阿托伐他汀对高胆固醇血症小鼠海马中tau蛋白磷酸化水平的影响

Objective To investigate the phosphorylation of tau protein and the effect of atorvastatin on tau protein phosphorylation in hypercholesteremic mouse hippocampus. Methods Male Kunming mice were randomly divided into normal control group, hypercholesteremia group, and low-dose atorvastatin treatment group (8mg·kg-1·d-1), middle-dose group (15mg·kg-1·d-1),and high-dose group (20mg·kg-1·d-1) with five mice in each group. The hypercholesteremia mouse model was induced by cholesterol-enriched diets. The expression level of tau protein phosphorylation in mouse hippocampus was detected by Western blot and immunohistochemistry methods. The activities of mitogen-activated protein kinase (MAPK) and cyelin dependent kinase 5 (Cdk-5) were measured by liquid scintillation counting of the hippocampus incorporated radioactivity and immunoprecipetation activity assay respectively. Results In hypercholesteremic group, the expression level of tau protein phosphorylation in mouse hippocampus was significantly increased(F=14.761,P＜0.01)as compared with control group, and so were MAPK activity and Cdk-5 activity(all P＜0.01). Atorvastatin treatment group showed the decreased expression level of tau protein phosphorylation at ser396/404 site in low-dose group (F=6.349,P＜0.05),middle-dose group (F=10.283,P＜0.01) and high-dose group (F=10.511,P＜0.01) as compared with hypercholesteremia mouse group. The activities of MAPK and Cdk-5 were decreased along with the increasing atorvastatin doses. Conclusions Hypercholesteremia induces tau protein hyperphosphorylation in mouse hippocampus. Abnormal cholesterol metabolism may play an important role in the pathology process of neurodegeneration in brain. Atorvastatin might inhibit tau protein hyperphosphorylation by inhibiting the activations of MAPK and Cdk-5 in brain, atorvastatin may have the protect effect for tau protein.     

阿托伐他汀对高胆固醇血症小鼠海马中tau蛋白磷酸化水平的影响

Objective To investigate the phosphorylation of tau protein and the effect of atorvastatin on tau protein phosphorylation in hypercholesteremic mouse hippocampus. Methods Male Kunming mice were randomly divided into normal control group, hypercholesteremia group, and low-dose atorvastatin treatment group (8mg·kg-1·d-1), middle-dose group (15mg·kg-1·d-1),and high-dose group (20mg·kg-1·d-1) with five mice in each group. The hypercholesteremia mouse model was induced by cholesterol-enriched diets. The expression level of tau protein phosphorylation in mouse hippocampus was detected by Western blot and immunohistochemistry methods. The activities of mitogen-activated protein kinase (MAPK) and cyelin dependent kinase 5 (Cdk-5) were measured by liquid scintillation counting of the hippocampus incorporated radioactivity and immunoprecipetation activity assay respectively. Results In hypercholesteremic group, the expression level of tau protein phosphorylation in mouse hippocampus was significantly increased(F=14.761,P＜0.01)as compared with control group, and so were MAPK activity and Cdk-5 activity(all P＜0.01). Atorvastatin treatment group showed the decreased expression level of tau protein phosphorylation at ser396/404 site in low-dose group (F=6.349,P＜0.05),middle-dose group (F=10.283,P＜0.01) and high-dose group (F=10.511,P＜0.01) as compared with hypercholesteremia mouse group. The activities of MAPK and Cdk-5 were decreased along with the increasing atorvastatin doses. Conclusions Hypercholesteremia induces tau protein hyperphosphorylation in mouse hippocampus. Abnormal cholesterol metabolism may play an important role in the pathology process of neurodegeneration in brain. Atorvastatin might inhibit tau protein hyperphosphorylation by inhibiting the activations of MAPK and Cdk-5 in brain, atorvastatin may have the protect effect for tau protein.     

Effect of bone marrow mesenchymal stem cells on the Smad expression of hepatic fibrosis rats

Objective:To investigate the impact of bone marrow mesenchymal stem cells on Smad expression of hepatic fibrosis rats.Methods:A total of 48 adult female SD rats were randomly divided into three groups,normal control group(n=10),observation group(n=19)with liver fibrosis model rats injected with BMSCs cells:model group(n=19),with liver fibrosis model rats injected with physiological saline.Serum index,TGF-β1 and Smad expression were detected.Results:TypeⅢprocollagen,Ⅳcollagen,hyaluronic acid,laminin levels of observation group were significantly lower than those of model group(P0.05).The content and expression of TGF-β1in serum and liver tissue of observation group were significantly lower than those of model group(P0.05).Compared with normal control group,the Smad3,Smad4 mRNA and protein expression of model group were significantly increased,the Smad7 mRNA and protein expression were significantly reduced(P0.05).Compared with model group.Smad3,Smad4 mRNA and protein expression of observation group were significantly reduced,and Smad7 mRNA expression were significantly increased(P0.05).Conclusions:BMSCs can regulate Smad expression to some extent,and reduce the degree of liver fibrosis.     

Effect of lentivirus-mediated integrin αVβ3-sh RNA on tumor growth of mice with lung cancer xenografts

Objective: To study the effect of lentivirus-mediated integrin αVβ3-sh RNA on tumor growth of mice with lung cancer xenograft. Methods: Lung cancer tissue, paracancer tissue and normal tissue were collected and integrin αVβ3 expression was detected; BALB/c nude mice were selected, divided into integrin αVβ3 knockdown group(KD group) and negative control group(NC group), and inoculated with cells stably infected by integrin αVβ3-sh RNA lentivirus and cells stably infected by negative control-sh RNA lentivirus respectively, the growth of tumor tissue was continuously observed, and the number of apoptosis cells as well as the expression of angiogenesis, apoptosis and invasion genes in tumor tissue were detected. Results: m RNA content and protein content of integrin αVβ3 in lung cancer tissue were significantly higher than those in paracancer tissue and normal tissue; increasing trend of tumor tissue volume of KD group was weaker than that of NC group, and tumor volume at various points in time of KD group was lower than that of NC group; m RNA contents and protein contents of VEGF, FGF, EGF, Bcl-2, MMP-9, MMP-12 and MMP-13 in tumor tissue of KD group were lower than those of NC group, and apoptosis index as well as m RNA content and protein content of Bax were higher than those of NC group. Conclusions: The expression of integrin αVβ3 increases in lung cancer tissue, and lentivirus-mediated integrin αVβ3-sh RNA can inhibit tumor growth of mice with lung cancer xenografts.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

S-phase delay in human hepatocellular carcinoma cells induced by overexpression of integrin β1

AIM: To clarify the mechanisms of integrin overexpression in negatively regulalting the cell cyde control of hepatocellular carcinoma cells SMMC-7721.METHODS: The cell cycle pattern was determined by flow cytometry. The mRNA and protein expression levels were assayed by RT-PCR and Western blot, respectively. Stable transfection was performed by Upofectamine 2000 reagent,and cells were screened by G418.RESULTS: Overexpression of α5β1 or β1 integrin induced S-phase delay in SMMC-7721 cells, and this delay was possibly due to the accumulaltion of cydin-dependent kinase inhibitors (CKIs) p21^cip1 and p27^kip1. The decrease of protein kinase B (PKB) phosphorylation was present in this signaling pathway, but focal adhesion kinase (FAK) was not involved.When phosphorylation of PKB was solely blocked by wortmannin, p27^kip1 protein level was increased. Moreover,S-phase delay was recurred when attachment of the parental SMMC-7721 cells was inhibited by the preparation of poly-HEME, and this cell cycle pattern was similar to that of β1-7721 or α5β1-7721 cells.CONCLUSION: S-phase delay induced by overexpression of integdn 151 subunit is attributed to the decrease of PKB phosphorylation and subsequent increases of p21^cip1 and p27^kip1 proteins, and may be involved in the unoccupied α5β1 because of lack of its ligands.     

Expression and significance of MMP-9 and MDM2 in the oncogenesis of lung cancer in rats

Objective:To observe the expression of matrix metalloproteinase-9(MMP-9)and mouse double minute 2 homolog(MDM2)in the oncogenesis of lung cancer in rats and to explore their clinical value.Methods:A total of 140 rats were selected,of which 20 were selected randomly as the control group;and the remaining 120 as the observation group.The observation group was injected with benzopyrene to establish diseases model such as tissue proliferation,abnormal proliferation and lung cancer.Delected the MMP-9 levels of lung tissue by enzyme-linked assay,detected the MDM2 levels of lung tissue by immunochemistry assay.Results:The MMP-9 and MDM2 expression of the lung cancer group and the abnormal proliferation group were significantly higher than that in the tissue proliferation group and the control group,the difference was significant(P0.05).And the MDM2 expression of the tissue proliferation group was significantly higher than that in the control group,the difference was significant(P0.05).There was no significant difference in the MMP-9 expression between the tissue proliferation group and the control group(P0.05).The MDM2 and MMP-9 expression were increased in turn in the small cell carcinoma,squamous cell carcinoma and adenocarcinoma,the difference was statistically significant(P0.05).The MMP-9 and MDM2 expressions of stageⅢand stageⅣlung cancer tissue in rats were significant higher than that during stageⅠand stageⅡ,the difference was significant(P0.05).There was no significantly different in the MMP-9 and MDM2 expressions between stageⅢand stageⅣ(P0.05),and there is no significant difference of the MMP-9and MDM2 expressions between stageⅠand stageⅡ(P0.05).Conclusions:The expression of MMP-9 and MDM2 in lung tissue was associated with lung disease and lung cancer,both of them may be involved in the development and metastasis of lung cancer.Combined detection can be used as therapy and prognostic indicators for lung cancer.     

Effect of Chaiqinchengqi decoction on sarco/endoplasmic reticulum Ca~（2＋）-ATPase mRNA expression of pancreatic tissues in acute pancreatitis rats

AIM： To investigate the effect of Chaiqinchengqi decoction （CQCQD） on sarco/endoplasmic reticulum Ca2＋-ATPase （SERCA） mRNA expression of pancreatic tissues in acute pancreatitis （AP） rats. METHODS： Thirty Sprague-Dawley （SD） rats were randomized into control group, AP group and CQCQD group （n = 3 × 10）. The rats in the CQCQD group were intragastrically administered with CQCQD （10 mL/kg every 2 h） after induction of AP by intraperitoneal injection of caerulein （50 μg/kg.h × 5） within 4 h. At 6 h after the induction of AP model, pancreatic tissues were collected for the pathological observation, mRNA extraction for determination of SERCA1 and SERCA2 mRNA expression or pancreatic acinar cell isolation for measurement of fluorescence intensity （FI） of intracellular calcium ion concentration [Ca2＋ i. RESULTS： There was no expression of pancreatic SERCA1 mRNA in the control group and the AP group. The expression of pancreatic SERCA2 mRNA in the AP group was down-regulated （expression ratio = 0.536; P = 0.001） compared with the control group, while that in the CQCQD group was up-regulated （expression ratio= 2.00; P = 0.012） compared with AP group. The FI of intracellular [Ca2＋ of pancreatic acinar cells in the AP group （138.2 ± 23.1） was higher than the C group （111.0 ± 18.4） and the CQCQD group （118.7 ± 15.2 ） （P 〈 0.05） and the pancreatic pathological score in the CQCQD group was lower than that in the AP group （5.7 ± 1.9 vs 9.2 ± 2.7, P 〈 0.05）.CONCLUSION： CQCQD can up-regulate the expression of SERCA2 mRNA of pancreatic tissues, reduce intracellular calcium overload and relieve pancreatic tissue lesions.     

Inhibitory effect of oxymatrine on hepatocyte apoptosis via TLR4/PI3K/Akt/GSK-3β signaling pathway

AIM To evaluate the effect of oxymatrine(OMT) on hepatocyte apoptosis in rats with lipopolysaccharide(LPS)/D-galactosamine(D-Gal N)-induced acute liver failure(ALF). METHODS LPS/D-Gal N was used to establish a model of ALF in rats. To evaluate the effect of OMT, we assessed apoptosis by transmission electron microscopy, and the pathological changes in the liver by light microscopy with hematoxylin and eosin staining. An automated biochemical analyzer was used to measure serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST). Enzyme-linked immunosorbent assay was used to determine the levels of tumor necrosis factor(TNF)-α and interleukin(IL)-1β. Western blotting was used to detect protein levels in liver tissues. Streptavidin peroxidase immunohistochemistry was used to observe expression of Toll-like receptor(TLR)4, active caspase-3, Bax and Bcl-2. RESULTS All rats in the normal control and OMT-pretreated groups survived. The mortality rate in the model group was 30%. OMT preconditioning down-regulated apoptosis of hepatocytes and ameliorated pathological changes in liver tissue. The levels of AST, ALT, TNF-α and IL-1β in the model group increased significantly, and were significantly reduced by OMT pretreatment. OMT pretreatment down-regulated expression of TLR4 and active caspase-3 and the Bax/Bcl-2 ratio, and upregulated expression of P-AktSer473(Akt phosphorylated at serine 473) and P-GSK3βSer9(glycogen synthase kinase 3β phosphorylated at serine 9) induced by LPS/D-Gal N. CONCLUSION OMT inhibits hepatocyte apoptosis by suppressing the TLR4/PI3K/Akt/GSK-3β signaling pathway, which suggests that OMT is an effective candidate for ameliorating acute liver failure.     

S-phase delay in human hepatocellular carcinoma cells induced by overexpression of integrin β1

AIM:To clarify the mechanisms of integrin overexpression in negatively regulating the cell cycle control of hepatocellular carcinoma cells SMMC-7721.METHODS: The cell cycle pattern was determined by flow cytometry. The mRNA and protein expression levels were assayed by RT-PCR and Western blot, respectively. Stable transfection was performed by Lipofectamine 2000 reagent,and cells were screened by G418.RESULTS: Overexpression of α5β1 or β1 integrin induced S-phase delay in SMMC-7721 cells, and this delay was possibly due to the accumulation of cyclin-dependent kinase inhibitors (CKIs) p21cip1 and p27kip1. The decrease of protein kinase B (PKB) phosphorylation was present in this signaling pathway, but focal adhesion kinase (FAK) was not involved.When phosphorylation of PKB was solely blocked by wortmannin, p27kip1 protein level was increased. Moreover,S-phase delay was recurred when attachment of the parental SMMC-7721 cells was inhibited by the preparation of polyHEME, and this cell cycle pattern was similar to that of β1-7721 or α5β1-7721 cells.CONCLUSION: S-phase delay induced by overexpression of integrin β1 subunit is attributed to the decrease of PKB phosphorylation and subsequent increases of p21cip1 and p27kip1 proteins, and may be involved in the unoccupied α5β1because of lack of its ligands.