Protective Effects Of Huayutongluo Traditional Chinese Medicine And Its Decomposed Recipes On Diabetic Nephropathy Rats And On The Expression Of Mas Reporter

Objective: Through fed with high fat diet and injection of streptozotocin(STZ) to develop diabetic nephropathy rats model, Setting up the normal control group(C), model group(M), Huayutongluo Chinese medicine group(Q), Huayu Chinese medicine group(H), Tongluo Chinese medicine group(T), Irbesartan group(I). By observing the general situation of rats, biochemical index,morphological changes of renal tissue,and the expression level of Mas receptor to explore the protective effect and the mechanism of Huayutongluo Chinese medicine,as well, the relationship between Mas receptor with diabetic nephropathy.Provides the basis for the clinical treatment of diabetic nephropathy.Methods: Fifty 75 male SD rats adaptively fed after a week were randomly selected 10 for the normal control group(C=10) and others belong to bulding group. Building group fed with high fat feed for 4 weeks. After fasting rats for 12 hours, Intraperitoneal injection of STZ 35 mg/kg. The rats showing random blood glucose ≥ 16.7 mmol/L 72 hours after injection were considered as successfully duplicated diabetic models. A total of 5 rats which death and not conform to the standard were eliminated. Building rats were randomly divided into model group(M group, n =12), given the correspond ing volume water by gavage. Intraperitoneal group(I=12), treated with irbesartan suspension to intervene. The whole Chinese medicine group(Q =12), were administered with Huayu and Tongluo Chinese medicine,compo sed of danshen, chuanxiong,quanxie,shuizhi, dilong formula particles.Tong luo group(T=12), composed of quanxie,shuizhi, dilong formula particles. Hua yu group(H=12) composed of danshen,chuanxiong formula particles. Given them the corresponding volume water by gavage. Rats′ urine was collected at the end of 16 th weeks.Animals were taked blood tests under anesthesia blood indicators, And to remain the kidney tissue for light microscopy, electron microscopy, immunohistochemistry, western blot, Real- Time PCR for detecting Mas receptor expression levels.Results: 1 The general situation of the experimental ratsThe rats of group C′color is gloss, muscle is plump,Their water intake, food intake and urine output is normal.But other groups appeared typically symptoms of diabete, such as polydipsia, polyphagia, polyuria and Loss of body weight. The above symptom group M is the most significant and the symptoms in treated groups were less than M group.Diabetes complications began to appear at 7 weeks after injection of STZ,such as cataract,diabetic foot. But the symptoms of treated groups have different degrees of ease during the process. 2 Weight, kidney weight and kidney hypertrophy indexThe result shows that: Rat kidney weight had no significant differences between groups(P>0.05);Compared with C, The body weight of M and treatment groups reduced significantly(P < 0.05), kidney hypertrophy index increased significantly(P < 0.05), Compared with M, the kidney hypertrophy index of I was obviously reduced(P < 0.05),There is no obvious difference between Q、T、H group with M group. 3 Quantitative of ptoteinuria in 24 hourCompared with group C, the total 24 hours urine protein levels of M group and treatment group were significantly higher(P < 0.05),But the treatment group was lower than that in group M.There is no obvious difference between I group and Chinese traditional medicine group(P > 0.05).4 Random blood glucoseCompared with group C, the random blood glucose levels in rats of group M and the treatment group was obviously increased(P < 0.05).There is no obvious difference between M group and treatment group(P > 0.05). 5 Other blood test results 5.1 PLTI, M group compared with group C, the platelet count increased significantly(P < 0.05), There is no obvious difference between C、Q、T、H group. 5.2 Liver functionTP and ALB level of C group were higher than other groups(P<0.05), TP level of I,Q,T group increased significantly than M group(P<0.05),but has no obvious difference between eath other(P>0.05).TP level between H and M group has no significant difference(P>0.05). The ALB level of M group was lower than that of treatment group(P<0.05), no significant difference between each treatment group(P > 0.05). 5.3 Renal functionCompared with group C, M group and each treatment group rats CR level increased significantly(P<0.05), and building groups have no obvious difference(P>0.05); The BUN level of Q,T,I group was obviously higher than that of C group, lower than that of group M(P<0.05), no obvious difference between H and M group(P>0.05). 5.4 Blood lipidCompared with group C, M group and each treatment group rats TG levels increased significantly(P<0.05), but the Q,T,H group was lower than that in group M(P<0.05), Rat TG level between I and M group had no significant difference(P>0.05); Compared with group C, rats TC levels of M group and each treatment group increased significantly(P<0.05), Between M group and each treatment group,rats TC level had no obvious difference(P>0.05);Compared with group C,M group and each treatment group rats LDL levels increased significantly(P<0.05),but the treatment group rats LDL level was lower than that in group M(P<0.05), LDL levels of Q group significantly decreased than group I(P<0.05), there is no significant difference between Q、T and H group(P>0.05). 5.5 HemagglutinationGroup Q, group T, H and group C,group M PT levels have difference(P< 0.05),significantly lower than that of group C,but significantly higher than the group M(P<0.05),No significant differences between group I and group M(P>0.05);M group and each treatment group rats compared with group C,INR levels significantly decreased(P<0.05),but Q group increased significantly than group M(P<0.05). 6 Kidney tissue morphology observation 6.1 Electron microscope observationGroup C in the rat kidney tissue structure is clear,thickening of glomerular basement without membrane,foot processes uniform distribution; Mesangial area without broadening.M owed clear group of renal tissue structure,glomerular basement membrane thickening of high homogeneity; Foot process fusion; Mesangial and matrix proliferated.Group I and Q,T, H structure is clear,the segmental thickening of glomerular basement membrane,part of foot process fusion,mesangial matrix increased slightly. Comparison between the treatment group,group H slightly heavier than other treatment group. 6.2 Light observationGroup C in the rat kidney tissue structure is clear, no obvious hypertrophy or atrophy,in glomerular.capillary lumens is clear and had no expansion, basement membrane had no thickening,mesangial without hyperplasia;.Glomerular of group M become hypertrophy,basement membrane thickened obviously,mesangial proliferated and mesangial area broadened. Compared with group M,the I and Q,T,H group pathological changes significantly reduced; But the group H compared to other treatment group,its pathological performance slightly heavier. 7 Immune histochemical method to detect the expression of renal tissue MasThe results showed: Mas are expressed in each group,it express in rat renal cortical tissue of proximal tubule and only a small expression within the glomeruli.Compared with group C,Mas expression of building groups in renal tissue of rats decreased significantly(P<0.05), compared with the group M, the expression of Mas in each treatment group was obviously increased(P< 0.05);Comparison between treatment groups,the expression of Mas of group H is less than other groups, group Q, T and I had no difference(P>0.05). 8 Real- Time PCR detection of renal tissue Mas m RNA expressionThe results showed: Compared with group C, Masm RNA expression of building groups in renal tissue of rats decreased significantly(P<0.05), Compared with the group M,the expression of Masm RNA in renal cortical at each treatment group was obviously increased(P<0.05);Group Q,T and group I had no significant difference(P>0.05). 9 Western blot method- detect the expression of Mas in renal corticalThe results showed: Compared with C, Mas expression levels of each building module in renal cortex were significantly lower(P<0.05); Compared with group M, the expression level of Mas in renal cortical of treatment group increased significantly(P<0.05); Group Q, T and group I had no significant difference(P>0.05).Conclusion:1 Combined with kidney hypertrophy index, 24 hours urinary protein and pathological changes of electron microscope and light observation, proved that diabetic rats become diabetic nephropathy after 16 weeks. DN rats had dark tail veins; and their blood in high viscosity, hypercoagulable state. These were in accordance with the features of blood-stasis-blocking-collaterals syndrome in Chinese medicine.2 Huayutongluo recipe and demolition as similar with irbesartan can improve the general situation of rats, reduce urinary protein, maintain plasma albumin content, reduce the serum creatinine,urea nitrogen,lighten the DN pathological changes to effect kidney protection.3 Huayutongluo traditional Chinese medicine and its demolition have an effective on reducing the number of platelets, regularing lipid metabolism and improving prothrombotic states. Compared three groups of traditional Chinese medicine group, the whole party and tongluo group therapy effect remarkablily, is better than that of huayu Chinese medicine group.4 Mas expression in DN rats renal cortex has obviously decreased. Huayutongluo Chinese medicine can improve the expression of Mas in renal cortical tissue of diabetic nephropathy rat,may play a role of renal protection for its one of the reasons.