Role of β2-adrenoceptor-β-arrestin2-nuclear factor-κB signal transduction pathway and intervention effects of oxymatrine in ulcerative colitis

Objective:To investigate the β2-adrenoceptor(β2AR)-β-arrestin2-nuclear factor-κB(NF-κB) signal transduction pathway and the intervention effects of oxymatrine in a rat model of ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups,which included the normal control group,the model group, the mesalazine group and the oxymatrine treatment group,with 10 rats per group.Experimental colitis induced with trinitrobenzene sulfonic acid(TNBS) was established in each group except the normal control group.The rats in the oxymatrine treatment group were treated with intramuscular injection of oxymatrine 63 mg/(kg·d) for 15 days and the rats in the mesalazine group were treated with mesalazine solution 0.5 g/(kg·d) by gastric lavage for 15 days. The rats in the normal control group and model group were treated with 3 mL water by gastric lavage for 15 days. Diarrhea and bloody stool were carefully observed.Histological changes in colonic tissue were examined on day 7 in 2 rats per group that were randomly selected.The expression of β2AR,β-arrestin2 and NF-κB p65 in colon tissue and spleen lymphocytes were detected with immunohistochemistry and Western immunoblotting techniques on day 16 after fasting for 24 h.Six rats died of lavage with 2 each in the normal control,the model group and the mesalazine group;and were not included in the analysis.Results:The rats in the model group suffered from looser stool and bloody purulent stool after modeling.But in the oxymatrine and mesalazine groups,looser stool and bloody purulent stool reduced after treatment.And the colonic wall in the model group was thickened and the colon length shortened.The colon mucosa was congested in multiple areas with edema,erosion,superficial or linear ulcer and scar formation,while the intestinal mucosa injury reduced in the mesalazine and oxymatrine groups(P<0.01).In colonic mucosa and in spleen lymphocytes,compared with the normal control group,the expression of NF-κBp65 were significantly increased(P<0.01) in the model group while the expressions ofβ2AR andβ-arrestin2 were significantly decreased(P<0.01).Compared with the model group,the expression of NF-κBp65 was significantly decreased in the mesalazine group(P<0.01) and oxymatrine treatment group(P<0.01) while the expressions of β2AR and β-arrestin2 were significantly increased(P<0.01).There were no statistically significant differences in the expression of β2AR,β-arrestin2 and NF-κBp65 between the mesalazine group and oxymatrine group(P>0.05).Conclusions:The β2AR-β-arrestin2-NF-κB signal transduction pathway participated in the pathologic course of ulcerative colitis.Oxymatrine attenuated ulcerative colitis through regulating the β2AR-β-arrestin2-NF-κB signal transduction pathway.     

Efficacy of Topical versus Oral 5-Aminosalicylate for Treatment of 2,4,6-Trinitrobenzene Sulfonic Acid-induced Ulcerative Colitis in Rats简

5-aminosalicylic acid（5-ASA） is drug of choice for the treatment of ulcerative colitis（UC）. In this study, the efficacy of topical versus oral 5-ASA for the treatment of UC was examined as well as the action mechanism of this medication. A flexible tube was inserted into the rat cecum to establish a topical administration model of 2,4,6-trinitrobenzene sulfonic acid（TNBS）-induced UC. A total of 60 rats were divided into sham operation group（receiving an enema of 0.9% saline solution instead of the TNBS solution via the tube）, model group, topical 5-ASA group, oral Etiasa group（a release agent of mesalazine used as positive control） and oral 5-ASA group（n=12 each）. Different treatments were administered 1 day after UC induction. The normal saline（2 mL） was instilled twice a day through the tube in the sham operation group and model group. 5-ASA was given via the tube in the topical 5-ASA group（7.5 g/L, twice per day, 100 mg/kg）, and rats in the oral Etiasa group and oral 5-ASA group intragastrically received Etiasa（7.5 g/L, twice per day, 100 mg/kg） and 5-ASA（7.5 g/L, twice per day, 100 mg/kg）, respectively. The body weight was recorded every day. After 7 days of treatment, blood samples were drawn from the heart to harvest the sera. Colonic tissues were separated and prepared for pathological and related molecular biological examinations. The concentrations of 5-ASA were detected at different time points in the colonic tissues, feces and sera in different groups by using the high pressure liquid chromatography（HPLC）. The results showed that the symptoms of acute UC, including bloody diarrhea and weight loss, were significantly improved in topical 5-ASA-treated rats. The colonic mucosal damage, both macroscopical and histological, was significantly relieved and the myeloperoxidase activity was markedly decreased in rats topically treated with 5-ASA compared with those treated with oral 5-ASA or Etiasa. The mRNA and protein expression of IL-1β, IL-6, and TNF-α was down-regulated in the colonic tissue of rats topically treated with 5-ASA, significantly lower than those from rats treated with oral 5-ASA or Etiasa. The concentrations of 5-ASA in the colonic tissue were significantly higher in the topical 5-ASA group than in the oral 5-ASA and oral Etiasa groups. It was concluded that the topical administration of 5-ASA can effectively increase the concentration of 5-ASA in the colonic tissue, decrease the expression of proinflammatory cytokines, alleviate the colonic pathological damage and improve the symptoms of TNBS-induced acute UC in rats.     

Effect of oxymatrine on hepatic gene expression profile in experimental liver fibrosis of rats

Objective:To investigate the effects of oxymatrine on hepatic gene expression profile in a rat model of liver fibrosis.Methods:Forty healthy male SD rats were randomly divided into three groups,a normal group（n=8）,a model group（n=16）,and an oxymatrine treatment group（n=16）.Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride（CCI₄）.The rats in the treatment group received oxymatrine via celiac injection at a dosage of 40 mg/kg once a day at the same time.The rats in the model and normal groups received saline at the same dosage via celiac injection.Serum levels of aspartate aminotransferase （AST）,alanine transarninase（ALT）,alkaline phosphatase（AKP）,hyaluronic acid（HA）,and laminin（LN）were assayed.The deposition of collagen was observed with HE and Masson staining.Effect of oxymatrine on hepatic gene expression profile was detected by oligonucleotide microarray analysis with Affymetrix gene chip rat U230A. Quantitative real-time polymerase chain reaction（QRT-PCR）was carried out to confirm the expression changes of six genes.Results:Oxymatrine significantly improved liver function,lowered serum levels of HA and LN,and decreased the degree of liver fibrosis,compared with the model group（P<0.05）.A total of 754 differentially expressed genes were identified by gene chip between the model group and the normal group,among which 438 genes increased and 316 genes decreased over two folds.Compared with the model group,86 genes were downregulated markedly in the oxymatrine group（P<0.05）,including collagen I and other genes related to extracellular material（ECM）,integrin signal transduction genes,early growth response factor genes,and proinflammatory genes;28 genes were upregulated significantly（P<0.05）,including cytochrome P450（CYP450） superfamily genes,glycolipids metabolism and biological transformation related genes.Six genes were confirmed with QRT-PCR,consistent with the result from microarray.Conclusion:Oxymatrine could affect the expression of many functional genes and may be useful in the prevention and treatment of liver fibrosis.     

Effect of qi-tonifying and stasis-eliminating therapy (益气活血法) on expression of vascular endothelial growth factor and its receptors Flt-1, Flk-1 in the brain of intracerebral hemorrhagic rats

Objective： To investigate the effects and mechanism of qi-tonifying and stasiseliminating （QTSE） therapy on the expression of vascular endothelial growth factor （VEGF） and its receptors Fit-1 and FIk-1 in the brains of intracerebral hemorrhagic （model） rats. Methods： One hundred and eighty Sprague-Dawley rats were randomly divided into six groups： the normal group （n=5）, the sham-operative （SO） group （n=35）, the model group （n=35）, the QTSE group （n=35）, the QT group （n=35） and the SE group （n=35）. All the rats except those in the normal group and SO group were established into an intracerebral hemorrhage（ICH） model by intracerebral injection of collagenase type Ⅶ and the latter three were orally administered with Buyang Huanwu Decoction （补阳还五汤, a classical recipe for QTSE） or with some of its components for qi-tonification and for stasis-elimination, respectively. To the other three groups, normal saline solutions were given instead. Behavioral tests were carried out in the animals randomly chosen from each group on days 1, 2, 4, 7, 14, 21 and 28 after modeling. The expressions of VEGF, FIk-1 and Fit-1 were determined by immunohistochemistry and the number of vascular segments with positive expression in the injured brain area of the rats was calculated. Results： From day 7 onwards, the asymmetric forelimb use rate in the QTSE group recovered more significantly than that in the other model groups. In the model group, the expressions of VEGF, FIk-1 and Fit-1 appeared on day 1 and reached a peak on day 21, then weakened gradually. In the QTSE group, as compared with the other model groups, a higher level of VEGF expression was shown from day 7 （P〈0.01） and a higher level of Fit-1 expression was shown from the 7th day to the 21st day （P〈0.01）. Conclusion： QTSE therapy can up-regulate the expressions of VEGF and its receptors （FIk-1 and Fit-l） and improve the recovery of kinetic function in the ICH rats, which may be correlated with its action in modulating vascular regeneration to promote the reconstruction of microvascular networks in the damaged areas.     

Effect of Qingre Buyi Decoction (清热补益汤) on Nitric Oxide and Histomorphology of Intestinal Mucosa in Rats with Radiation Enteritis

Objective: To explore the mechanism of Qingre Buyi Decoction (清热补益汤,QBD) in prevention and treatment of radiation enteritis in rats. Methods: Forty-eight Wistar rats were randomly divided into four groups, the TCM group, the WM group, the model group and the control group, 12 in each group. Rats in the former three groups were given orally with QBD, norfloxacin and normal saline once a day for 7 successive days, after being irradiated with X-ray at single dose of 10 Gy for modeling of radiation enteritis, while rats in the control group were untreated. Animals were sacrificed at the end of the medication. NO concentration, mean height and number of villi per centimeter in their small intestinal mu-cosa were measured. Results: The intestinal NO concentration was significantly lower in the TCM and WM groups than that in the model group(P<0.05), while the number of villi was significantly more and the height higher in the former two groups than those in the model group (P<0.01 for both), but no signif     

左归丸和右归丸对实验性自身免疫性脑脊髓炎大鼠轴突再生过程中脑源性神经营养因子表达及环磷酸腺苷/蛋白激酶A信号转导通路的作用

Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA）signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis（EAE）,in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods：The rats were randomly divided into normal control（NC）,model（MO）,prednisone acetate（PA）,ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein（MBP）68-86.The brain and spinal cord were harvested on the 14th and 28th day postimmunization（PI）,the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction（PCR）,and the cAMP levels were detected by using enzyme-immunoassay method.Results：（1）On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group（P〈0.05 or P〈0.01）.（2）On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group（P〈0.05 or P〈0.01）,and the cAMP level in YGP group was higher than that in ZGP group（P〈0.05）.The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively（P〈0.01）.（3）On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively（P〈0.05）.（4）On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions：The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mec     

Combined Transplantation of Neural Stem Cells and Olfactory Ensheathing Cells Improves the Motor Function of Rats with Intracerebral Hemorrhage

Objective To investigate the effects of combined transplantation of neural stem cells （NSC） and olfactory ensheathing cells （OEC） on the motor function of rats with intracerebral hemorrhage. Methods In three days after a rat model of caudate nucleus hemorrhage was established, NSCs and OEC, NSC, OEC （from embryos of Wistar rats） or normal saline were injected into bematomas of rats in combined transplantation group, NSC group, OEC group, and control group, respectively. Damage of neural function was scored before and in 3, 7, 14, 30 days after operation. Tissue after transplantation was observed by immunocytochemistry staining. Results The scores for the NSC, OEC and co-transplantation groups were significantly lower in 14 and 30 days after operation than in 3 days after operation （P〈0.05）. The scores for the NSC and OEC groups were significantly lower than those for the control group only in 30 days after operation （P〈0.05）, while the difference for the NSC-OEC group was significant in 14 days after operation （P〈0.05）. Immunocytochemistry staining revealed that the transplanted OEC and NSC could survive, migrate and differentiate into neurons, astrocytes, and oligodendrocytes. The number of neural precursor cells was greater in the NSC and combined transplantation groups than in the control group. The number of neurons differentiated from NSC was significantly greater in the co-transplantation group than in the NSC group. Conclusion Co-transplantation of NSC and OEC can promote the repair of injured tissue and improve the motor fimction of rats with intracerebral hemorrhage.     

Monocyte chemotactic protein 1 increases homing of mesenchymal stem cell to injured myocardium and neovascularization following myocardial infarction

Objective：To investigate the effect of MCP-1 on mesenchymal stem cells（MSCs） homing to injured myocardium in a rat myocardial infarction（MI） model. Methods：Rat myocardial infarction model was established by permanent left anterior descending branch ligation. Mesenchymal stem cells from donor rats were cultured in IMDM and labeled with BrdU. The Rats were divided into two groups. Monocyte chemotactic protein I（MCP-1） expression were measured by in situ hybridization and immunohistochemistry in the sham operated or infarcted hearts at 1, 2, 4, 7, 14 and 28 days post operation in MCP-1 detection group. The rats were injected with MCP-1, anti-MCP-1 antibody or saline 4 days after myocardial infarction in intervention group. Then, a total of 5 × 10^6 cells in 2.5 ml of PBS were injected through the tail vein. The number of the labeled MSCs in the infarcted hearts was counted 3 days post injection. Cardiac function and blood vessel density were assessed 28 days post injection. Results：Self-generating MCP-1 expression was increased at the first day, peaked at the 7^th day and decreased thereafter post MI and remained unchanged in sham operated hearts. The MSCs enrichment in the host hearts were more abundant in the MI groups than that in the non-MI group（P= 0.000）, the MSCs enrichment in the host hearts were more abundant in the MCP-1 injected group than that in the anti-MCP-1 antibody and saline injected groups （P = 0.000）. Cardiac function was improved more in MCP-1 injected group than anti-MCP-1 antibody and saline injected groups（P= 0.000）. Neovascularization in MCP-1 injected group significantly increased compared with that of other groups（P = 0.000）. Conclusion： Myocardial MCP-1 expression was increased only in the early phase post MI. MCP-1 may enhance MSCs homing to the injured heart and improve cardiac function by promoting neovascularization.     

Effect of Tiantai No.1(天泰1号)on Gene Expression Profiles in Hippocampus of Alzheimer's Disease Rats by Bioinformatic Analysis

Objective:To study the effect of Tiantai No.1(天泰1号) on gene expression profile in hippocampus of Alzheimer's disease(AD) rat,molecular genetic target points of the effect of this drug were defined,its molecular genetic pharmacodynamic mechanism of anti-AD was further explored at molecular gene level,and a scientific basis was provided for its clinical availability and promotion.Methods:Thirty male SpragueDawley rats were divided into three groups with 10 rats per group:sham-operation group,model group and Tiantai No.1 group.Sterile surgical procedure was applied,the model group with bilateral hippocampal injection of Aβ_(1-40) was established,and normal saline was used instead of Aβ_(1-40)in the sham-operation group.One week after the models was made,rats were administered by gastric lavage once every day for three consecutive weeks.The rats of the sham-operation group and the model group were daily fed with purified water by lavage;the rats of the Tiantai No.1 group treated group were administered with Tiantai No.1 by lavage.Total RNAs of hippocampus tissues were extracted with Trizol,the changes of hippocampus gene expression profiles in the above three groups were analyzed by using Affymetrix rat whole genome expression profile microarray.Results:Microarray analysis showed that,compared with the sham-operation group,the hippocampus of the model group had 50 up-regulated genes with significant difference(fold change 2),and 21 down-regulated genes with significant difference(fold change 0.5);compared with the hippocampus of the model group,the hippocampus of the Tiantai No.1 group was found to have 5 up-regulated genes with significant difference(fold change 2) and 20down-regulated genes with significant difference(fold change 0.5).The functions of differentially expressed genes of the groups were involved in nervous system's development,neuronic differentiation and function-regulation,cellular growth and differentiation and apoptosis,synaptic occurrence and plasticity,inflammation and immune response,ion channels/transporters,cellular signal transduction,cellular material/energy metabolism and so on.Conclusion:Tiantai No.1 can regulate hippocampal function,and further regulate the brain function of animals in multiple gene target points by a number of ways.     

Effect of Kangshuai Yizhi Formula I (抗衰益智方 I) on learning and memory dysfunction induced by scopolamine in mice

Objective：To evaluate the improvement of Kangshuai Yizhi FormulaⅠ（抗衰益智方Ⅰ,KYFⅠ）on the learning and memory dysfunction in mice,and on the mechanism of the hippocampal cholinergic system and the nervous system of monoamine which are closely related to learning and memory function.Methods：Mice in the low-,middle-,and high-dose KYFⅠgroups were given low-,middle-,and high-dose KYF,respectively, by gastrogavage for 35 successive days.Animals in the control group and the model group were treated with distilled water.The acute learning and memory dysfunction model was established by injection of scopolamine from day 31,and Morris water maze was used to assess the behavior performance of scopolamine-induced model mice for five days.The activities of acetylcholinesterase（AChE）,choline acetyl transferase（ChaT） and the content of monoamine neurotransmitters in hippocampus were measured.The activity of monoamine oxidase（MAO）in hippocampus and serum was also detected.Results：（1）Compared with the control group,the mean escape latency was shortened,and the frequency across the platform and the staying time at the platform area on the 5th day were decreased in the model group by Morris water maze test.The activities of AChE and MAO were increased,and the ChaT activity and monoamine neurotransmitter content were decreased as well.（2） The escape latency for 4 days in the low-,middle-,and high-dose KYFⅠgroups was significantly shortened than that in the model group,with the shortest latency in the high-dose KYFⅠgroup（P〈0.05,P〈0.01）.The frequency across the platform was significantly increased and the staying time at the platform was significantly prolonged in the middle-and high-dose KYFⅠgroups（P〈0.05,P〈0.01）.（3）As compared with the model group,the activity of ChaT and the content of monoamine neurotransmitters in the hippocampus were significantly increased, and the activities of AchE and MAO were significantly decreased in the hippocampus in the high-dose KYFⅠgroup（P〈0.01）.Conclusions：High-dose KYFⅠcan significantly improve the learning and memory dysfunction induced by scopolamine in mice.Its mechanism may be related to improving the central cholinergic system and regulating the hippocampal monoamine neurotransmitters.     

The Active Metabolite of Leflunomide A771726 Inhibits Corneal Neovascularization

The effects of A771726, the active metabolite of leflunomide, on experimental rat corneal neovascularization （NV） in vivo and on cultured human umbilical vein endothelial cells in vitro were studied. The corneal NV was induced by alkali burn in 40 SD rats. The rats were randomly divided into 4 groups with 10 rats in each group. Group A was treated with 0.9% sodium chloride （control group）, and group B, group C and group D were given different concentrations of A771726 eye drops （0.5%, 1.0%, 2.0% respectively） 4 times daily during days 0--28. The occurrence and development of corneal NV were observed at 4, 7, 14, 21 and 28 day after alkali burn by a slit lamp microscope. The cultured human umbilical vein endothelial cells （ECV-304） were incubated with A771726 solution at different concentrations （20, 40, 80, 160, 320 μmol/L） for 36 h. The proliferation of cells was assessed by methyl thiazolyl tetrazolium （MTT）, and the expression of proliferating cell nuclear antigen （PCNA） in cells was detected by using immunofluorescence under the laser confocal microscope. The rat model showed that the onset of corneal NV was delayed and progression of corneal NV was inhibited in the groups C and D. The corneal NV areas in groups C and D were significantly smaller than in groups A and B （P〈0.01）. No significant difference was found in corneal NV areas between groups C and group D （P〉0.05）. A771726 solution （940 μmol/L） could inhibit proliferation of human umbilical vein endothelial cells and decrease the expression of PCNA in cells significantly, A771726, as the active metabolite of leflunomide, strongly prevented corneal NV induced by alkali burn in the in vivo model, and inhibited proliferation of human umbilical vein endothelial cells in the in vitro model. Therefore, A771726 may serve as an angiogenic inhibitor in the treatment of corneal NV.     

Effects of tanshinone II A on the myocardial hypertrophy signal transduction system protein kinase B in rats

Objective： To study the effect of tanshinone ⅡA on the cell signal transduction system protein kinase B （Akt） in rats with hypertrophy of the myocardium induced by partial constriction of the thoracic aorta. Methods： Rat models of myocardial hypertrophy were established by the thoracic aorta partial constriction method. Forty-eight rats were randomly divided into the sham-operative group, the model group, the valsartan treatment group, and the low-, medium-, and high-dose tanshinone treatment groups. The heart mass index （HMI）, left ventricular mass index （LVMI）, ejection fraction （EF）, left ventricular posterior wall （LVPW）, and interventricular septal thickness （IVS） were detected by high-frequency ultrasonography. The myocardial fiber diameter （MFD） was detected by HE staining, and the contents of p-Akt and p-Gsk3β in the myocardium were detected by Western blot. Results： Compared with the sham-operative group, the levels of HMI, LVMI, LVPW, IVS, and MFD were increased respectively in the other groups （P〈0.05）; the contents of p-Akt and p-Gsk3β were also increased in the other groups. Compared with the model group, the levels of HMI, LVMI, LVPW, IVS, and MFD were decreased respectively in all treatment groups （P〈0.05）; the contents of p-Akt and p-Gsk3β were decreased in all treatment groups as well. There was no significant difference, however, among the low-, medium-, and high-dose tanshinone treatment groups and the valsartan treatment group （P〉0.05）. Conclusion： Tanshinone HE A can prevent myocardial hypertrophy by its action on the protein kinase B （Akt） signaling pathway.     

Iloprost inhibits fracture repair in rats

Background Previous studies have shown that prostaglandins （PGs） dramatically stimulate healing processes in bone.However,the effect of prostaglandin l2 （PGI2） on fracture healing remains unclear.To investigate the effect of PGI2,a study on fracture healing process in closed tibia fractures was designed.Methods Thirty-six Sprague-Dawley male rats were randomized into two groups.On the first day,their right tibias were fractured by three-point bending technique.The study group （n=18） received a single injection of 10 μg/kg iloprost for 5 days,while the control group （n=18） received saline solution in the same way.On the 7th,14th and 28th days following the fracture,six rats were sacrificed and their right legs were harvested in each group.The progression of fracture healing was assessed for each specimen by the scores of radiography （by Lane-Sandhu） and histology （by Huo et al）.Results On the 7th day,the radiographic and histologic scores were equal.On the 14th day radiographic total score was 6 and histologic total score was 23 in the iloprost group,whereas radiographic total score was 11 and histologic total score was 33 in the control group.On the 14th day radiographic and histologic scores were significantly decreased in the iloprost group compared to the control group （P 〈0.05）.On the 28th day radiographic total score was 12 and histologic total score was 37 in the iloprost group,whereas radiographic total score was 15 and histologic total score was 40 in the control group.On the 28th day although there was a decrease in radiographic and histologic scores of the iloprost group acording to control group,it was not statistically significant （P 〉0.05）.Conclusion Iloprost delays fracture healing in early stage in rats.     

Effect of Wumeiwan on Cytokines TNF-α, IL-6, IL-8, IL-10 and Expression of NF-κBp65 in Rats with Ulcerative Colitis

The effects of Wumeiwan （WMW） on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis （UC） were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine （SASP） was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley （SD） rats were randomly divided into four groups （n=14 in each group, with equal ratio of male and female）： normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene （DNCB） immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α, IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased （P〈0.01）, while those of IL-10 significantly reduced （P〈0.01） after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group （P〈0.05）. The levels of IL-6, IL-8, and TNF-α were lower, while the level oflL-10 was higher in WMW group than in SASP group. NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group （P〈0.01）, and there was significant difference between WMW group and SASP group （P〈0.05）. It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.     

Effects of Curcumin on Pain Threshold and on the Expression of Nuclear Factor κB and CX3C Receptor 1 after Sciatic Nerve Chronic Constrictive Injury in Rats

Objective：To investigate the effects of curcumin on pain threshold and the expressions of nuclear factorκB（NF-κB）and CX3C chemokine receptor 1（CX3CR1）in spinal cord and dorsal root ganglion（DRG）of the rats with sciatic nerve chronic constrictive injury.Methods：One hundred and twenty male Sprague Dawley rats,weighing 220-250 g,were randomly divided into 4 groups.Sham surgery（sham）group：the sciatic nerves of rats were only made apart but not ligated;chronic constrictive injury（CCI）group：the sciatic nerves of rats were only ligated without any drug treatment;curcumin treated injury（Cur）model group：the rats were administrated with curcumin 100 mg/（kg·d）by intraperitoneal injection for 14 days after CCI;solvent control（SC）group：the rats were administrated with the solvent at the same dose for 14 days after CCI.Thermal withdrawal latency（TWL）and mechanical withdrawal threshold（MWT）of rats were respectively measured on pre-operative day 2 and postoperative day 1,3,5,7,10 and 14.The lumbar segment L4-5 of the spinal cord and the L4,L5 DRG was removed at post-operative day 3,7 and 14.The change of nuclear factorκB（NF-κB）p65 expression was detected by Western blotting while the expression of CX3CR1 was determined by immunohistochemical staining.Results：Compared with the sham group,the TWL and MWT of rats in the CCI group were significantly decreased on each post-operative day（P〈0.01）,which reached a nadir on the 3rd day after CCI,and the expressions of NF-κB p65and CX3CR1 were markedly increased in spinal cord dorsal horn and DRG.In the Cur group,the TWL of rats were significantly increased than those in the CCI group on post-operative day 7,10 and 14（P〈0.05）and MWT increased than those in the CCI group on post-operative day 10 and 14（P〈0.05）.In addition,the administration of curcumin significantly decreased the positive expressions of NF-κB p65 and CX3CR1 in spinal cord and DRG（P〈0.05）.Conclusion：Our study suggests that curcumin cou     

Shenling Baizhu Powder (参苓白术散) Ameliorates Pi (Spleen)-Deficiency-Induced Functional Diarrhea in Rats

Objective To explore the mechanism of Pi(Spleen)-deficiency-induced functional diarrhea(FD)model rats treated by Shenling Baizhu Powder(参苓白术散,SBP).Methods Thirty male Sprague-Dawley rats were randomly divided into 5 groups including control,model,low-,medium-,and high-dose SBP groups(SBPLDG,SBPMDG,SBPHDG),6 rats in each group,respectively.Pi-deficiency-induced FD rats model was developed through Radix et Rhizoma Rhei gavage for 7 days.After modeling,the rats were treated with 3 doses of SBP[0.93,1.86,and 3.72 g/(kg·d)],and the rats in the control and model groups were given pure water for 7 days.The diarrhea index was calculated.On the 7th and 14th days,the traveled distance of rat was measured by the open field test.Serum D-xylose content was determined by the phloroglucinol method and interleukin(IL)-10 and IL-17 levels were measured using an enzyme-linked immunosorbent assay kit.The content of Treg cells was determined by flow cytometry.Results Compared with the control group,the diarrhea index and IL-17 level in the model group were significantly higher and the total exercise distance and D-xylose content significantly decreased(P>0.05).The expression of IL-10 in the SBPHDG group was significantly up-regulated,and serum D-xylose level and Treg cells increased significantly compared with the model group(P>0.05).Conclusion High-dose SBP exhibited ameliorating effects against Pi-deficiency induced FD,which might be attributed to its modulations on intestinal absorption function as well as adaptive immunity in mesenteric lymph nodes of rat.     

Effects of Zuogui Pill (左归丸) on Wnt Singal Transduction in Rats with Glucocorticoid-induced Osteoporosis

Objective: To reveal the mechanism of Zuogui Pill (左归丸) in treatment of glucocorticoid-induced osteoporosis from the angle of the Wnt signal transduction pathway and to provide further experimental evidence for expounding the scientific connotation of "the kidney dominating the bones" in TCM. Methods: Forty-two male Wistar rats were selected and randomly divided into three groups, control group (n=12), model group (n=15) and Zuogui Pill group (n=15). Form the beginning, The rats were injected dexamethasone for eight weeks to make the model of osteoporosis, and the Zuogui Pill were administered intragastrically to the rats of Zuogui Pill group for eight weeks. The relative morphological parameters were measured in the undecalcified tibial slices. And the protein expression levels of Wnt1, LRP-5 and β-catenin in rat tibial osteoblasts (OB) and bone marrow stromal cells (BMC) were detected by immunohistochemistry. Results: Compared with the control group, TBV% and TFS% decreased significantly, while TRS% increased significantly, and the protein expression of Wnt1, LRP-5 and β-catenin in OB and BMC decreased significantly in the model group. And compared with the model group, TBV% and TFS% increased significantly, and expression levels of Wnt1, LRP-5 and β-catenin proteins increased significantly in the Zuogui pill group. Conclusion: Zuogui Pill can prevent and treat glucocorticoid-induced osteoporosis in rats by up-regulating the expression of the key signal molecules Wnt1, LRP-5 and β-catenin in Wnt signal transduction pathway.     

加味竹叶石膏汤及其成分可预防大鼠放射性食管炎的发生

Objective:To investigate the effect of Modified Zhuye Shigao Decoction(加味竹叶石膏汤,MZSD)and its components on preventing radiation esophagitis of rats.Methods:One hundred Wistar rats were randomly divided into 5 groups,including the control group,radiation model group,MZSD group,Zhuye Shigeo Decoction(竹叶石膏汤,ZSD)group,and added Ingredients group,20 rats in each group.The model of radiation esophagitis of rat was established by once local radiation of 40 Gy(330 Mulmin)with a high energy linear accelerator.The administration of Chinese medicine was continued for 14 days from 7 days before radiation application in the three treatment groups.On the 7th and 14th day,the serum was isolated and the levels of inflammatory cytoldnes tumor necrosis factor(TNF-α),interleukin 1 13(IL-1β)and IL-8 were tested.The pathological slices of esophagus were obtained,and the pathological changes were observed.During the whole process,weight and food intake were recorded each day.Results:On the 7th day after radiation,the esophagus of rats in the MZSD group was almost intact,and the pathological injury score was significantly lower than that of the radiation model group,ZSD group and added ingredients group(P0.01).Compared with the control group,the body weight and food intake of rats in the radiation model group were significantly decreased,and the levels of TNF-α,IL-1β and IL-8 were significantly increased(P0.05 or P0.01),while the MZSD group showed a significant increase in body weight and food intake,and a significant decrease in the levels of TNF-α,IL-1β and IL-8 compared with the radiation model group,ZSD group and added ingredients group(P0.05 or P0.01).Conclusion:MZSD prevents the development of radiation esophagitis probably by inhibiting the generation and release of the inflammatory cytoldnes TNF-α,IL-1β and IL-8.     

Effects of Zuogui Pill(左归丸) and Yougui Pill(右归丸) on the Expression of Brain-Derived Neurotrophic Factor and Cyclic Adenosine Monophosphate/Protein Kinase A Signaling Transduction Pathways of Axonal Regeneration in Model Rats with Experimental Autoimm

Objective:To study the effects of Zuogui Pill(左归丸,ZGP)and Yougui Pill(右归丸,YGP)on the expressions of brain-derived neurotrophic factor(BDNF)and cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis(EAE),in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods:The rats were randomly divided into normal control(NC),model(MO),prednisone acetate(PA),ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein(MBP)_(68-86).The brain and spinal cord were harvested on the 14th and 28th day postimmunization(PI),the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction(PCR),and the cAMP levels were detected by using enzyme-immunoassay method.Results:(1)On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group(P0.05 or P0.01).(2)On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group(P0.05 or P0.01),and the cAMP level in YGP group was higher than that in ZGP group(P0.05).The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively(P0.01).(3)On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively(P0.05).(4)On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions:The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.