| Objective: To observe the effect of Traditional Chinese medicine Berberine Hydrochloride on the expression of Toll like receptor 4(Toll like receptor 4,TLR4)and its downstream signaling mediated monocyte chemoattractant protein(Monocyte chemoattractant protein-1,MCP-1)on thoracic aorta tissue of diabetic rats,and then clarify the effect of berberine on Diabetic macrovascular disease and its mechanism.Methods: 50 adult male Wistar rats were placed in a free drinking water environment and fed for 1 weeks.50 adult male Wistar rats were randomly divided into two groups: normal control group(NC,n=10)and diabetes mellitus(DM)group(n=40).The rats in DM group were injected STZ60mg/kg one-time into enterocoelia(STZ before use with 0.1M with the concentration of 1% sodium citrate buffer solution,p H =4.2)and the rats in normal control group were injected same volume of sodium citrate buffer.After 72 hours,the rats whose blood glucose?16.7mmol/L had chosen as diabetic rats.20 of 30 diabetes rats were randomly divided into the follow group: DM,BBR group(n=10).The rats in BBR group was given the100g/(kg body weight/d)Berberine Hydrochloride medicine with intragastric administration.The rats in NC and DM groups have given the same volume of saline.All of rats drink and eat freely in the experiment.After 8 weeks,fasting12 h with 3% pentobarbital(80mg/kg)intraperitoneal injection of anesthesia rapidly,used in rats to harvest blood and thoracic aorta tissue.Blood was used for the determination of cholesterol(TC),triglyceride(TG),alanine aminotransferase(ALT),aspartate transaminase(AST),blood urea nitrogen(BUN),serum creatinine(SCr)levels.The thoracic aorta fixed with 4%paraformaldehyde,Paraffin embedded,sliced,for HE staining to observe thepathological changes of blood vessels.The expression levels of TLR4 and MCP-1 thoracic aortic tissue were examined with immunohistochemical staining.The positive area and integrated optical density was measured by colour image analytic system.The measurement data were described by——X ±S.The comparison of each group before and after intervention using matching t test.The one-way ANOVA used for test the difference between each group.Kruskal-Wallis H test is used when variance is notneat.All the data were processed with SPSS 21.0 statistical processing software and inspection level:?=0.05.Results:1 The change of the general condition of rats Normal control group of rats in good mental state,urine and drinking water is moderate,normal stool,hair even shiny,eyes have God,responsive,lively and moving.The rats of DM group and BBR group compared with the normal group: mental state is poor,urine volume increased,water quantity increased,defecation increased,the colour dark matt,unresponsive,reduced activity;2 The change of blood glucose Before the treatment,DM group and BBR group blood glucose were significantly higer than normal control group(P<0.05);Before the treatment,DM group and BBR group compared with each other has no obvious difference(P>0.05).After the treatment,DM group and BBR group blood glucose were higer than normal control group(P<0.05);After the treatment,DM group and BBR group compared with each other has no obvious difference(P>0.05).There were no difference of blood glucose before and after the treatment in each group(P>0.05);3 The effect on blood lipidThe effect on cholesterol: After the treatment,compared with the normal control group,DM group and BBR group cholesterol have no obvious difference(P>0.05);DM group and BBR group compared with each other cholesterol has no obvious difference(P>0.05);The effect on triglyceride: After the treatment,compared with the normal control group,DM group and BBR group triglyceride have no obviousdifference(P>0.05);DM group and BBR group compared with each other triglyceride has no obvious difference(P>0.05);4 The impact on alanine aminotransferase,aspartate transaminase,blood urea nitrogen and serum creatinineAfter the treatment,compared with the normal control group,DM group and BBR group alanine aminotransferase,aspartate transaminase,blood urea nitrogen and serum creatinine have no obvious difference(P>0.05);DM group and BBR group compared with each other alanine aminotransferase,aspartate transaminase,blood urea nitrogen and serum creatinine has no obvious difference(P>0.05);5 The Expression of the vascular tissue morphology After HE staining,Observe each rat thoracic aorta morphological changes under the light microscope : The blood vessels form of NC group is normal,endothelial cells arranged closely,membrane is integrated,no inflammatory cells and amorphous material deposition;The intimal artery of DM group is visible incrassated and incomplete,part of the lining to the cavity,foam cells and cranny is visible,smooth muscle cells are disordered;The intimal artery of BBR group is visible thickening,part of the endothelial cells disappeared,foam cells can be seen occasionally;6 The expression of TLR4 in thoracic aorta After lavage treatment for8 weeks,compared with normal control group,the expression of TLR4 was significantly the expression of TLR4 of BBR group(P<0.05);Compared with DM group,BBR group significantly decreased(P<0.05);7 The expression of MCP-1 in thoracic aorta After lavage treatment for 8 weeks,compared with normal control group,the expression of MCP-1was significantly increased in DM group and BBR group(P<0.05);Compared with DM group,the expression of MCP-1 of BBR group significantly decreased(P<0.05).Conclusion:1 Berberine had no effect on blood glucose,blood lipid,alanine aminotransferase,aspartate aminotransferase,blood urea nitrogena nitrogenand creatinine levels.2 In the normal control group,HE staining in thoracic aorta was normal,without atherosclerosis,the morphology of the thoracic aorta in the diabetic rats and the rats treated with berberine hydrochloride suggested that arterial sclerosis occurred in HE staining.The model of atherosclerosis was successfully replicated in diabetic rats.3 The expression of TLR4 and MCP-1 in the thoracic aorta of the normal group was lower than that in the diabetic group and the berberine hydrochloride group,indicating that it was involved in the occurrence and development of vasculopathy.4 Berberine hydrochloride can inhibit the expression of TLR4 and MCP-1 in the thoracic aorta of diabetic rats,and has antiatherosclerotic effect.5 The anti atherosclerosis effect of berberine hydrochloride is not achieved by lowering blood glucose and blood lipid. |
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