Up-regulation of HIF-1α and VEGF Expression by Elevated Glucose Concentration and Hypoxia in Cultured Human Retinal Pigment Epithelial Cells

Summary： In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF）, human RPE cells were cultured in 5,56 mmol/L glucose （control group）, 5.56 mmol/L glucose with 150 !a mol/L COCl2 （hypoxic group）, 25 mmol/L glucose （high glucose group） and 25 mmol/L glucose with 150 μmol/L COCl2 （combination group）. RT-PCR was used to detect the expression of HIF-1α and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1α and VEGF proteins. Although the small amount of HIF-1α protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1α mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1α mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1α, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1α of RPE cell, and HIF-1α protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF-1α protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy.     

Abdominal Fat Accumulation with Hyperuricemia and Hypercholesterolemia Quail Model Induced by High Fat Diet

Objective To establish abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model fed with high fat diet. And then to investigate the pathological characteristics of this quail model. Methods Thirty Longcheng quails were randomly divided into two groups： control group and model group （n=15）. The control group quails were fed with normal diet and model group quails were fed with high fat diet for 14 days. After a 12-hour overnight fast, liver and abdominal fat at euthanasia as well as serum were collected. The levels of serum uric acid, total cholesterol, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C）, triglyceride, free fatty acid （FFA）, and blood glucose were assayed. The activity changes of adenosine deaminase （ADA）, xanthine oxidase （XOD）, lipoprotein lipase （LPL）, hepatic lipase （HL）, and fatty acid synthetase （FAS） were analyzed. Results Compared with control group, the abdominal fat content （0.74±0.63 vs. 1.36±0.65 g, P〈0.05） and abdominal fat index （0.44%±0.30% vs. 0.85%±0.30%, P〈0.01） as well as live lipid index （3.61%±0.65% vs. 11.33%±2.14%, P〈0.01） in model group significantly increased; the levels of serum uric acid （210.61±94,76 vs. 304.25±141.94 /amol/L, P〈0.05）, total cholesterol （4.20±0.51 vs. 20.10±11.25 mmol/L, P〈0.01）, LDL-C （1.16±0.29 vs. 10.78±6.48 mmol/L, P〈0.01）, and FFA （0.39±0.14 vs. 0.55±0.15 mmol/L, P〈0.01） in model group significantly increased; HDL-C （5.85±0.95 vs. 4.14±2.03 mmol/L, P〈0.05） significantly decreased; the levels of triglyceride and blood glucose had no significant changes （P〉0.05）; the activities of ADA （9.71±3.05 vs. 17.19±5.10 U/ml, P〈0.01） and XOD （10.58±6,88 vs. 19.22＋9.44 U/L, P〈0.01） in model group significantly increased; and FAS, LPL, HL had no significant changes （P〉0.05）. Conclusions High fat diet can induce abdominal fat accumulation with hyperuricemia and hypercholesterolemia quail model. The changes of uric acid and lipid metabolic enzyme activities may he the pathological mechanism of abdominal fat accumulation with hyperuricemia and hypercholesterolemia.     

Effect of ARB on Expression of CD68 and MCP-1 in Adipose Tissue of Rats on Long-term High Fat Diet

In adipose tissue of rats on long-term high fat diet, the inflammatory changes the roles of angiotensin receptor blocker （ARB） in pimelitis and insulin resistance （IR） were observed. LR rat model was established by feeding high calorie and high fat diet. The change in insulin sensitivity was detected by euglycemic-hyperinsulinemic clamp technique 8 weeks after intervention by valsartan. The expression levels of CD68 and MCP-1 mRNA and proteins in adipose tissue were examined by RT-PCR and immunohistochemistry respectively. The parameters of blood glucose, insulin and blood lipid were analyzed. The results showed that in high fat diet group intra-abdominal obesity developed the content of visceral fat and the number of inflammatory cells in local adipose tissue were significantly increased （P〈0.01）, the levels of serum triglyceride, free fatty acids and fasting serum insulin were markedly increased, the insulin sensitivity was significantly lowered （P〈0.01）, and the expression of CD68 and MCP-1 was significantly increased as compared with control group （P〈0.01）. In ARB interventional group, the content of visceral fat, the number of inflammatory cells and the ex- pression of CD68 and MCP-1 in local adipose tissue were significantly reduced （all P〈0.01）, but the insulin sensitivity was significantly enhanced （P〈0.01） as compared with high fat diet group. There were pimelitis and IR in rats with obesity induced by long-term high calorie and high fat diet. The ARB can significantly inhibit the infiltration of macrophages and the expression of MCP-1 in adipose tissue, thereby attenuating the inflammation and improving LR in rats.     

Changes of High Mobility Group box 1 in Serum of Pig Acute Hepatic Failure Model and Significance

The role of the high mobility group box 1 （HMGB-1） in acute hepatic failure and the effect of artificial liver support system treatment on HMGB-1 level were investigated. Pig models of acute hepatic failure were induced by D-galactosamine and randomly divided into two groups with or without artificial liver support system treatment. Tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） levels were detected by the enzyme linked immunosorbent assay （ELISA）, the expression of HMGB-1 by Western blot, and serum levels of HMGB-1, liver function and hepatic pathology were observed after artificial liver support system treatment. The levels of TNF-α and IL-1β were increased and reached the peak at 24th h in the acute hepatic failure group, then quickly decreased. The serum level of HMGB-1 was increased at 24th h in the acute hepatic failure group and reached the peak at 48th h, then kept a stable high level. Significant liver injury appeared at 24th h and was continuously getting worse in the pig models of acute hepatic failure. In contrast, the liver injury was significantly alleviated and serum level of HMGB-1 was significantly decreased in the group treated with artificial liver support system （P〈0.05）. It was suggested that HMGB-1 may participate in the inflammatory response and liver injury in the late stage of the acute liver failure. Artificial liver support system treatment can reduce serum HMGB-1 level and relieve liver pathological damage.     

Effect of Huanglian Jiedu Decoction (黄连解毒汤) on glucose transporter 4 expression in adipose and skeletal muscle tissues of insulin resistant rats

Objective： To investigate the effects of Huanglian Jiedu Decoction （黄连解毒汤, HLJDD） on glucose transporter 4 （GLUT4） protein expressions in insulin-resistant murine target tissues. Methods： The experimental male Wistar rats were established into insulin resistant models by injecting streptozotocin （STZ 30 mg/kg） via caudal vein and feeding them with high fat high caloric diet, and randomly divided into the model group, the aspirin group and the HLJDD group. Besides, a normal group was set up for control. Changes of body weight （BW）, levels of serum fasting blood glucose （FBG）, serum fasting insulin （FINS） and oral glucose tolerance test （OGTT） were routinely determined. The expression of GLUT4 protein in adipose and skeletal muscle tissues before and after insulin stimulation was determined with Western blot. Results： In the HLJDD group after treatment, BW and FBG got decreased, OGTT improved, and the expression and translocation of GLUT4 protein elevated obviously, either before or after insulin stimulation, as compared with those in the model group, showing significant differences respectively. Conclusion： The mechanism of improving insulin resistance by HLJDD is probably associated with its effect in elevating GLUT4 protein expression and translocation in adipose and skeletal muscle tissues of insulin resistant rats.     

Effect of short term intensive multitherapy on carotid intima- media thickness in patients with newly diagnosed type 2 diabetes mellitus

Background Controlling plasma glucose levels, blood pressure and lipid levels is proven to reduce the risk of vascular complications in patients with type 2 diabetes mellitus. This has prompted intensive multitherapy targeted at several macrovascular risk factors. Carotid intima-media thickness （cIMT） is a reliable measure of early atherosclerosis. We sought to determine whether a 6-month intensive mutiltherapy program resulted in better goal attainment than usual care and its effect on the development of cIMT among patients with newly diagnosed type 2 diabetes mellitus.
Methods The study randomly assigned 220 patients with newly diagnosed type 2 diabetes mellitus to intensive or traditional therapy groups. The clinical parameters, such as fasting plasma glucose, total cholesterol, triglyceride, blood pressure, body weight and insulin were assessed at the baseline and after the 6-month therapy, cIMT of the patients was also obtained.
Results The average levels of fasting plasma glucose, hemoglobin Alc, total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C） in the intensive group were significantly lower than those in the control group at the end of 6-month treatment. By 6 months, a higher proportion of patients in the intensive therapy group than in the control group attained goals for fasting plasma glucose （FPG）, TC, LDL-C and hemoglobin Alc. With intensive multherapy the level of carotid intima-media thickness in the intensive therapy group was lower than that in the control group （（0.88±0.26） mm vs （0.96±0.22） mm, P 〈0.01）.
Conclusions The evidence from this clinical trial demonstrates that intensive glucose, lipid and blood pressure control in patients with newly diagnosed type 2 diabetes is associated with diabetic macrovascular benefits. Intensive multitherapy allows more patients to achieve aims of control and may reduce macrovascular complications and delay disease progression.     

Adipose tissue resistin gene expression in DIO and DR rats

Objective： To investigate the expression of resistin gene in diet-induced obesity （DIO） and diet resistance （DR） rats. Methods： DIO and DR models were prepared with male SD rats after 6 weeks feeding by a diet of relatively high fat, sucrose, and caloric content （HE diet）. Body-weight, fat mass, and the concentration of serum insulin were measured, and the expression of resistin and Peroxisome proliferator-activated receptory-7（PPAR-7） gene in whit adipose tissue （WAT） was also detected by RT-PCR. Results： ① Body weight, fat mass and the concentration of serum insulin were significantly increased in DIO rats and decreased in DR rats. ② The expression of resistin and PPAR7 gene was upregulated in DIO group and supressed in DR group, but the expression of resistin was not detectable in all samples within three groups. Conclusion： Resistin may serve as a link between obesity and insulin resistance, but the individual difference is enormous.     

Ethanol Extract of Purple Sweet Potato Tubers （Ipomoea batatas L） Decreases Blood Glucose and Increase Total Antioxidant Level in Rats with High Glucose Intake

High blood glucose level can increase the production of free radicals and the AGEs. Flavonoids from fruits and vegetables can prevent that events because they have antioxidant and hypoglycemic effects. The aim of this study was to examine possible hypoglycemic and antioxidant effect of the ethanol extract of purple sweet potato tubers. Subject of this study were 20 male adult rats divided into 4 groups with randomized pre-test and post-test control group design. Before treatment, blood samples were collected from retro-orbital sinus of all rats for blood glucose, MDA and total antioxidant measurement as pre-test data. After pre-test, group 1 （control group） rats were given normal diet without ethanol extract of purple sweet potato tubers. Group 2, 3, and 4 rats were given normal diet and ethanol extract of purple sweet potato tubers 1 cc/day, 2 cc/day, 4 cc/day, respectively, for one week. After one week of treatment, treatment groups were given single dose of high glucose intake. Over 1, 2, and 3 hours of treatment with high glucose, the blood samples were collected for post-test examination. The data were then analysed by one way anova and paired-t test, and the result showed a significant increase of blood glucose and MDA in group 1 （p 〈 0,01）, significant decrease of total antioxidant in group 1 （p 〈 0.01）. In group 3, all of the parameters were different with group 1 and group 2 （p 〈 0.01）. In group 4, MDA decreased significantly after treatment and the total antioxidant was increased significantly （p 〈 0.01）. From these findings, it can be concluded that ethanol extract of purple sweet potato tubers can maintain the blood glucose level within normal limit, prevent the increase of MDA, and increase total antioxidant of the blood after high glucose intake in rats. The effective dose of the extract in this research was medium dose.     

Clinical Observation on Electroacupuncture Treatment of 30 Cases of Chronic Hepatitis B

Objective: To probe into the methods and therapeutic effects of electroacupuncture (EA) on hepatitis B. Methods: 60 hospitalized cases of mild or moderate hepatitis B were randomly divided into a treatment group (30 cases) and a control group (30 cases). EA was applied at the points selected according to the differentiation of symptoms and signs. The treatment was given once daily for 30 minutes, and 2 weeks of treatments constituted a therapeutic course, with 2-3 courses on average in the treatment group. Conventional treatment for liver protection was given in the control group. Results: As compared with the control group, the duration of the treatment for recovery of the hepatic functions was significantly shorter, and the IL-8 level significantly lower in the treatment group. Conclusion: In treating hepatitis B, EA is effective in improving symptoms, recovering hepatic functions, and regulating immune functions to certain extent.     

Effects of Yixin Jiangya Capsules on Insulin Resistance and Tumor Necrosis Factor-α in Cases of Primary Hypertension with Left Ventricular Hypertrophy

Objectives: To observe the effects of Yixin Jiangya Capsules (益心降压胶囊 capsules for nourishing the heart and lowering blood pressure) on insulin resistance (IR) and tumor necrosis factor-α (TNF-α) in patients with primary hypertension with left ventricular hypertrophy (LVH). Methods: Totally 93 cases were randomly divided into a control group of 31 cases taking Enalapril and a treatment group of 62 cases taking Enalapril and Yixin Jiangya Capsules. Results: Fasting serum insulin (FSI) and TNF-α obviously increased and insulin sensitive index (ISI) significantly decreased in both groups before treatment as compared to those of a healthy group. After treatment, FSI, TNF-α and fasting blood glucose (FBG) obviously decreased and ISI remarkably increased in the treatment group, while ISI significantly increased and TNF-α obviously decreased in the control group. The curative effect in the treatment group was remarkably superior to that in the control group. FSI was positively related to TNF-α before treatment in both groups. Conclusion: FSI and TNF-α obviously increase and ISI significantly decreases in patients with primary hypertension with LVH. FSI and TNF-α influencing each other are involved in the generation and development of hypertension. Yixin Jiangya Capsules can improve IR and decrease TNF-α.     

Effect of Prepared Rhubarb on Insulin Resistance in Patients With Pregnancy Induced Hypertension

Objective: To investigate the effect of prepared rhubarb on insulin resistance in patients with pregnancy induced hypertension (PIH) and its mechanism. Methods: All the 92 patients accepted 75 g oral glucose tolerance test (OGTT) and insulin release test before and after treatment. These patients were divided into two groups (treated group and control group). Prepared rhubarb and nifedipine were given to the treated group, while nifedipine was given to the control group alone. Circulating endothelial cell (CEC), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) were measured and glucose area under curve (GAUC), insulin area under curve (IAUC), insulin sensitivity index (ISI) were calculated. And 30 normal pregnant women were selected as the healthy group. Results: CEC, TNF-a, IL-6, GAUC and IAUC of PIH patients were significantly higher than those of the normal healthy group; while ISI of PIH patients was significantly lower than that of the healthy group. ISI was significantly negatively correlat     

Effects of different dose endothelin-1 on expession of peroxisome proliferator-γ in cultured vascular smooth muscle cells of adult rats

Objective： To investigate the effects of endothlin-1 （ET-1） on vascular smooth muscle cells（VSMCs） proliferation and the expression of Peroxisome proliferator-activated receptorγ （PPAR-γ） in VSMCs. Methods： VSMCs of 16-week-old wistar rats thoracic aorta were cultured. VSMCs were treated by ET-1 for 48 h and observed of the proliferation by MTr. The expression of PPAR-γ mRNA and protein in cultured VSMCs treated by different concentration of ET-1 for 48 h was detected by RT-PCR and Western blot. Results： Compared with control group, VSMCs treated by ET-1 proliferated with the increase concentration of ET-1. There was significant differences among different groups （P 〈 0.01）. Meanwhile,the expression of PPAR-γ both in mRNA and in protein levels deceased. The expression of PPAR-γ in VSMCs was gradually decreased along with the increase concentration of ET-1. There was significant differences among different groups （ P 〈0.01）. Compared with control group, the expression of both PPAR-γ mRNA and PPAR-γ in ET-1 treated groups were lower（ P 〈 0.01 ）. Conclusion： ET-1 could induce VSMCs proliferation and the expression of PPAR-γ in VSMCs, which demonstrates that high dose ET-1 obviously weakens the function of PPAR-γ to increase VSMCs proliferation.     

Effects of Replenishing Qi, Promoting Blood Circulation and Resolving Phlegm on Vascular Endothelial Function and Blood Coagulation System in Senile Patients with Hyperlipemia

Objective： To observe the curative effect of the method of replenishing qi, promoting blood circulation and resolving phlegm on senile hyperlipemia and its effects on vascular endothelial function and blood coagulation system. Method： 96 patients with senile hyperlipemia were randomly divided into a treatment group and a control group. 56 patients in the treatment group orally took Danshen Jueming Granules （丹参决明颗粒） while 40 patients in the control group orally took Xuezhikang Capsules （血脂康胶囊）. Observed were indexes of blood lipid, vascular endothelial function, blood coagulation system and safety. Results： After treatment, the treatment group was obviously superior to the control group （P〈0.05） in reducing plasma total cholesterol （TC） and low density lipoprotein cholesterol （LDL-C） as well as in the ratio of thromboxane B2 （TXBz） to 6-keto-prostaglandin Flα （6-keto-PGF1α）, D-dimer （D-D） and fibrinogen （FIB）. Conclusion： Danshen Jueming Granules have the effect of regulating metabolism of blood lipid, and improving vascular endothelial function and blood coagulation system in senile patients with hyperlipemia.     

Prevalence of Abnormity of Blood Lipid and Associated Factors in Health Examination Population in Beijing

Objective To investigate the prevalence of abnormity of blood lipid and associated factors in healthy population in Beijing. Methods Totally, 38462 individuals who received health examination were enrolled in our study. We divided them into eight groups according to their ages. The levels of serum total cholesterol, triglyceride, high density lipoprotein cholesterol, and low density lipoprotein cholesterol were tested, and the relationship of blood lipid abnormity with body mass index （BMI） and fasting blood glucose was analyzed. Results The incidences of hypercholesterolemia, hyperglyceridemia, low high-density lipoprotein cholesterolemia, and hyper low-density lipoprotein cholesterolemia presented increasing trend in this population. The incidence rate of abnormity of blood lipid in health examination population increased with BMI increase. The incidence of abnormity of blood lipid in overweight and obesity population was significantly higher than that in low weight and normal weight populations （P〈0.05）. Meanwhile, the trend of abnormal blood lipid incidence coincided with that of abnormal fasting blood glucose. Conclusions The prevalence of overweight, obesity, and abnormity of blood lipid in Beijing presents increasing trend. The incidence of abnormity of blood lipid increases with BMI increase, in coincidence with that of fasting blood glucose.     

Protective effects of Modified Linggui Zhugan Decoction combined with short-term very low calorie diets on cardiovascular risk factors in obese patients with impaired glucose tolerance

OBJECTIVE:To observe the protective effects of modified Linggui Zhugan Decoction combined with short-term very low calorie diets(VLCDs) on cardiovascular risk factors in obese patients with impaired glucose tolerance(IGT).METHODS:Eighty-five obese patients with IGT of spleen hypofunction and dampness accumulation type were randomly divided into treated group(n=45)and control group(n=40).The treated group was given Linggui Zhugan Decoction combined short-term VLCDs.The control group was given basic weight-reduction treatment.The total course was 6 months.Changes of fasting plasma glucose(FPG),2-hour postprandial blood glucose(2hPG),glycosylated hemoglobin(HbA1c),fasting insulin(FINS),lipid metabolism,blood pressure,body mass index(BMI) and waist circumference(WC) were observed,and the outcomes were reviewed after the treatment.RESULTS:Compared with the control group,the levels of FPG,2hPG,HbA1c,FINS,HOMA-IR decreased significantly(P<0.05),and the indexes of total cholesterol(TC),triglyceride(TG),low-density lipoprotein(LDL),systolic blood pressure(SBP),diastolic blood pressure(DBP),BMI and WC were improved significantly(P<0.05) in the treated group.The transformation rate of normal glucose tolerance(NGT) was higher in treatment group than control group(P<0.01).No severe adverse reaction was found in the therapeutic course.CONCLUSION:The modified Linggui Zhugan Decoction combined with short-term VLCDs has good protective effects on cardiovascular risk factors as a treatment intervention for IGT with obesity,as it could improve glycometabolism,significantly decrease the levels of blood pressure,blood lipids,BMI,WC and effectively cut-off and reverse the development of diabetes mellitus.     

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.     

Combined use of extended-release niacin and atorvastatin: safety and effects on lipid modification

Background Cholesterol-lowering therapy with statins has been reported to reduce the morbidity and mortality of cardiovascular diseases. This study aimed to investigate the effects of combined application of extended-release niacin and atorvastatin on lipid profile modification and the risks of adverse events in patients with coronary artery disease. Methods Consecutive 108 patients with coronary artery disease and serum total cholesterol （TC） 〉 3.5 mmol/L were randomized into two groups： group A using atorvastatin and group B using extended-release niacin （niacin ER） and atorvastatin. Plasma lipid profile, glucose, and adverse events were assessed at the hospitalization, and 6 and 12 months after treatment. In addition, clinical cardiovascular events were evaluated after 12 months of treatment. Results The levels of TC, low density lipoprotein cholesterol （LDL-C） were significantly decreased （P 〈0.05） in groups A and B, but the levels of high density lipoprotein cholesterol （HDL-C） and ApoA increased by 29.36% and 40.81% respectively after 12 months of treatment in group B （P 〈0.01）. The medications were generally well tolerated in the two groups. No significant difference of adverse events was found between the two groups （group A： 3.2% vs group B 5.1%, P 〉0.05）. Conclusions Combined use of extended-release niacin with atorvastatin was superior to atorvastatin monotherapy alone in lipid profile regulation. Combination therapy with niacin ER and atorvastatin was well tolerated and safe in patients with coronary artery disease.     

Effect of FK506 on Expression of Hepatocyte Growth Factor in Murine Spinal Cord Following Peripheral Nerve Injury

This study is to investigate the effect of FK506 on expression of hepatocyte growth factor （HGF） in rats＇ spinal cord following peripheral nerve injury and to elucidate the mechanisms for neuroprotective property of FK506. Fifty male rats were randomly divided into normal group, injury group and treatment group. Models of peripheral nerve injury were established by bilateral transection of sciatic nerve 0.5 cm distal to piriform muscle. Then the treatment group received subcutaneous injection of FK506 （1 mg/kg） at the back of neck, while the injury group was given 0.9% saline. The L4-6 spinal cords were harvested at various time points after the surgery. Western blotting and immunofluorescent staining were used to detect the level and position of HGF in spinal cord. Immunofluorescent staining showed that HGF-positive neurons were located in anterior horn, intermediate zone and posterior horn of gray matter in normal spinal cord. Western blotting revealed that there was no significant difference in the expressions of HGF between the injury group and the normal group, while the expression of HGF was significantly higher in the treatment group than in the injury group 7 and 14 days after surgery. It is suggested that peripheral nerve injury does not result in up-regulation of the expression of HGF in spinal cord, while FK506 may induce high expression of endogenous HGF after injury thereby protecting neurons and promoting axonal outgrowth.     

Expression of cyclooxygenase-2 and its pathogenic effects in nonalcoholic fatty liver disease

Objective:To investigate the expression of cyclooxygenase-2 and its pathological effect in the experimental nonalcoholic fatty liver of rats,and to explore its possible mechanism.Methods:The rat NAFLD model was established by giving a fat-enriched diet.The blood samples were obtained form abdominal aorta and the levels of serum ALT,AST and IL-1,changes in the hepatic tissue 6-k-PGF1α TXB2 were measured.The expression level of COX-2 in rats livers were assayed by immunohistochemistry,RT-PCR and Western-blot.Results:Light microscope analysis revealed that hepatocytes were injured in the model group and slightly in the treatment group.The levels of serum TXB2 and IL-1 in the fatty liver rats were increased.Compared with the model group,the IL-1 and TXB2 increased significantly(P < 0.05),on the contrary,compared with the normal group,the hepatic tissue 6-Keto-prostagland decreased significantly in the model group(P < 0.05),the treatment group also increased but P > 0.05.There was no positive expression of COX-2 in hepatic tissue of normal rats.In the model group,there was positive expression of COX-2 antigen and the number of COX positive cells progressively increased at 4,8,12 wks.The intensity of expression of COX-2 had significantly increased(P < 0.05)and the intensity of COX-2 expression in the treated group decreased remarkably compared with the model group(P < 0.05).The expression of COX-2 mRNA and the level of COX-2 protein were significantly stronger in the liver of model rats compared with normal rats,and significantly weaker in treated rats,than in 8W and 12W model rats(P < 0.05).Conclusion:The increase of COX-2 expression in NAFLD is closely associated with the severity of liver inflammation and damage.COX-2 may play an important role in the progression of rat NAFLD,and the expression of COX-2 mRNA is downregulated by cyclooxygenase-2 inhibitor,which can depress the oxidative stress and control inflammatory response efficiently.     

Isoflavone Regulates Lipid Metabolism via Expression of Related Genes in OVX Rats Fed on a High-fat Diet

Objective To investigate the effects of isoflavone on body weight, fat mass, and gene expression in relation to lipid metabolism. Methods Thirty-six female SD rats were ovariectomized or sham-operated and fed on a high-fat diet. Two months later, abdominal incision was made, blood was collected to separate serum, and the liver and adipose tissue were immediately collected and weighed. Some portions of these tissues were frozen in liquid nitrogen and stored at -80℃. Results Ovariectomy （OVX） with a high-fat diet could induce obesity in rats, while treatment with isoflavone significantly inhibited the increase in body weight and fat mass in abdomen. Serum total cholesterol and leptin were significantly decreased in isoflavone group, compared with the OVX group. The mRNA expression of liver fatty acid synthase （FAS） in the OVX group was significantly higher than that in sham-operated group, while this difference was not observed in the isoflavone group. The mRNA expression of liver hormone-sensitive lipase （HSL） in the OVX rats tended to be lower than that in the sham-operated rats. Furthermore, a large amount of isoflavone maintained the mRNA expression at a sham level. Conclusion Isoflavone may prevent obesity induced by ovariectomy with a high-fat diet, in part by modulating gene expression related to lipid metabolism.