| Objective: This study develops DM rat animal model throughing high-sugar high-fat feed combined with intraperitoneal injection of a low dose of streptozotocin(STZ), and application of Xiezhuojiedu Huoxuetongluo traditional Chinese medicine, irbesartan and the both of drugs intervene the animal model, by observing the experimental animal’s general conditions,biochemical indexes, renal pathological changes and the expression level of Angiopoietin-like protein 2(Angptl 2), to investigate the renal protective effect and possible mechanism of Xiezhuojiedu Huoxuetongluo traditional Chinese medicine on DN rat.Methods: 85 healthy male Sprague-Dauley rats, weighing 170±10g, was fed adaptively one week. 15 rats were randomly selected as the normal group(ZG), given normal diet, the other 70 as module, given the high-sugar high-fat diet for 12 weeks after giving one-time intraperitoneal injection of STZ 35mg/kg, and normal group rats were injected with the same dose of 0.1 mol/l citrate buffer. 72 hours later, the model of diabetes was successfully built,which random blood sugar ≥16.7mmol/l, two rats died and one rat’s blood sugar not up to standard in the process of building, so they were removed. The successfull model rats were randomly divided into four groups: model group(MG) 17, traditional Chinese medicine group(ZG) 17, western medicine group(JG) 17, Chinese medicine and western medicine group(JG)16. normal group and model group were given amout of saline, each treatment group were given drug lavage, monitoring body weight, blood glucose and urine protein during the dosing period of 4 weeks, 8 weeks, 12 weeks. At the end of 16 th weeks, 12 hours of fasting, but water, collected 24-hour urine of each rat to detect 24 hours urinary protein, 10% chloral hydrate anesthetized rats, chose blood in abdominal aorta to measure biochemical indexs, picked the right kidney and weighed, tiked the renal tissue for observing the renal pathological morphological changes with light microscope,electron microscope and the expression of Angptl 2 in immunohistochemistry,detecting the contents of Angptl 2 m RNA. Results:Results: 1 The general conditions of experimental ratsNormal rats were in condition of good mental state, normal diet, hair luster, responsive and weight gradually increased. Module of rats all became more dranking, eating, urinating, apathetic, thin, fatigue, thin and so on, the symptoms in model group were severe.In the process of buliding module, two rats injected STZ died within 72 hours,one rat’ blood sugar was not up to standard, so they were rejected. In the process of drug delivery, four rats died in model group, one rat died in traditional Chinese medicine group, two rats died in western medicine group, at the same time, these rats’ blood sugar were not up to standard, which two in traditional Chinese medicine group, one in western medicine group, one in Chinese medicine and western medicine group, they were taken as well.2 The comparison of rats’ body weigh, kidney weight and the kidney hypertrophy index(kidney weigh/body weight) in each groupCompared with normal group, the weight of treatment group and model group was significantly lighter(P<0.05); the body weight was no significantly different in the treatment group and model group(P>0.05). Compared with normal group, the kidney hypertrophy index of the treatment group and model group was significantly higher(P<0.05); the kidney hypertrophy index between model group and treatment group was no significantly different(P>0.05). The comparison of kidney weight had no obvious difference in each group(P>0.05).3 The comparison of rats’ 24 h urinary total protein(24UTP) in each groupCompared with normal group, the 24 UTP was significantly increased in model group and each treatment group(P<0.05); compared with model group, each treatment group was significantly reduced(P<0.05); the comparison in each treatment group, Chinese medicine and western medicine group was low than traditional Chinese medicine group and western medicine group(P<0.05); there was no significant difference in traditional Chinese medicine group and western medicine group(P>0.05).4 The comparison of rats’ glycosylated hemoglobin(Hb A1c)Compared with normal group, the other group rats’ HbA1 c higher significant(P<0.05); the model group and western medicine group were higher than traditional Chinese medicine group, Chinese medicine and western medicine group(P<0.05); the model group and western medicine group had no significant difference(P>0.05); the traditional Chinese medicine group, Chinese medicine and western medicine group had no obvious difference(P>0.05).5 The comparison of rats’ total cholesterol(TC) and triglyceride(TG)Compared with normal group, the treatment groups and the model group TC,TG leves were significantly increased(P<0.05); compared with model group, the treatment groups TC,TG decreased significantly(P<0.05); comparison among the treatment groups, Chinese medicine and western medicine group TC,TG lower than the traditional Chinese medicine group and western medicine group(P<0.05); there was no statistical significantance between traditional Chinese medicine group and western medicine group(P>0.05).6 The comparison of rats’ serum total protein(TP) and albumin(ALB)Compared with normal group, the lever of TP and ALB in the model group and treatment groups were significantly lower(P<0.05); compared with model group, the treatment groups were higher(P<0.05); comparison among the treatment groups, Chinese medicine and western medicine group was higher than traditional Chinese medicine group,western medicine group(P<0.05); there was no obvious difference about TP and ALB in traditional Chinese medicine group and western medicine group(P>0.05).7 The comparison of rats’ blood urea nitrogen(BUN) and serum creatinine(Scr) BUN and Scr of normal group, model group, traditional Chinese medicine group, western medicine group, Chinese medicine and western medicine group had no significant differentance(P>0.05).8 The observation of renal pathology morphology8.1 Light microscopyThe renal tissue structure was clear and normal in normal group, glomerular structure had no abnormal, glomerular basement membrane and mesangial matris were normal without thickening; model group rats apprared glomerular hypertrophy, glomerular basement membrane and mesangial matris thickening, mesangial cell proliferation, foot process fusion. compared with model group, the renal pathology changes in each treatment group was significantly lighter.8.2 Electron microscopyThe normal group rats’ renal tissue structure was clear and normal, glomerular basement membrane thickening did not been seen, foot process arranged in order manner without fusion; glomerular basement membrane thickening and not uniform, mesangial matris increased, foot process fusion widely han been seen in model group; each treatment group’ pathology change compared with the model group was lighter, the difference among the treatment groups was not significant.9 Detecting the expression of Angptl 2 in renal tissue with immunohistochemistryA small amount of expression of glomerular Angptl 2 or no obvious expression in normal group; compared with the normal group, the expression of Angptl 2 increased obviously in the buliding module groups(P<0.05); compared with the model group, the expression of Angptl 2 reduced obviously in the treatment groups(P<0.05); compared with traditional Chinese medicine group and western medicine group, Chinese medicine and western medicine group’ expression was low(P<0.05); there was no significant difference between traditional Chinese medicine group and western medicine group(P>0.05). 10 Detecting the expression of Angptl 2 m RNA in renal tissue with Real-Time PCRCompared with the normal group, Angptl 2 m RNA expression significantly increased in model group and each treatment group(P<0.05); compared with the model group, the Angptl 2 m RNA expression of each treatment group significantly reduced(P>0.05); compared with the treatment groups, Chinese medicine and western medicine group Angptl 2 m RNA expression was less than traditional Chinese medicine group and western medicine group(P<0.05); There were no significant difference between the traditional Chinese medicine group and western medicine group(P>0.05)Conclusions: 1 Xiezhuojiedu Huoxuetongluo traditional Chinese medicine can significantly reduce 24 h urinary protein quantity, increase plasma protein, reduce blood sugar, improve blood lipid metabolism disorders and reduce the pathological change, meanwhile, it has a role in protecting kidney.2 Xiezhuojiedu Huoxuetongluo traditional Chinese medicine can downregulate the expression of Angptl 2, suggesting that it is one of possible mechanism of treatmenting diabetic nephropathy. |
PDF Full Text Request