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Zuogui hypoglycemic Yishen Decoction on mice with type 2 diabetic nephropathy podocyte structure and nephrin, podocin expression

Author: YinHongAi
Tutor: YuZuo
School: Hunan University of Traditional Chinese Medicine
Course: Chinese and Western medicine combined with clinical
Keywords: diabetic nephropathy MKR mice ZuoGui JiangTangYishen Decoction podocyte nephrin podocin
CLC: R285.5
Type: Master's thesis
Year: 2014
Downloads: 3
Quote: 0
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Abstract


Objective To explore the possibility of progressive diabetic nephropathy in mice model by high fat diet feed and unilateral nephrectomy and investigate the effects of Zuogui Jiangtang Yishen decoction (ZGJTYS) on the structure of podocyte and expression of nephrin, podocin in podocyte of MKR mice with diabetic nephropathy (DN).Methods Firstly, establish the model of diabetic nephropathy:Sixty transgenic MKR mice (8weeks old) who expressed dominant-negative IGF-1receptor and insulin receptor in skeletal muscle were divided randomly into three groups as follows:ordinary feed group, high fat feed group, high-fat diet feed and mono-nephrectomy model group. The body weight, fasting blood glucose (FBG), micro albumin of urine (UmAlb) were detected at at1st,2nd,4th,6th,8th week. Eight weeks later, all mice were put to death, then blood and kidney were prepared to detect the serum BUN and Cr and observe the pathologic changes. Secondly, the part of drug intervention:Forty MKR mice (8weeks old) were divided randomly into four groups as follows:negative control group (group A), DN model group(group B), ZGJTYS group(group C) and positive control group(group D, Gliquidone and benazepri). All mice from group B, C and D were received high-fat diet feed and unilateral nephrectomy. Four weeks after operation, all mice were received drug intervention, and four weeks later, all mice were put to death. The levels of micro albumin of urine (UmAlb) were measured with double-antibody sandwich ELISA, the serum BUN and Cr with biochemical, and the fasting blood glucose (FBG) with electrochemical detection method. The morphological structure changes of the podocytes were observed with transmission electron microscopes. The nephrin and podocin mRNA expression were measured by RT-PCR, the protein expression by western blotting.Results The part of establishing the model of diabetic nephropathy:As compared with ordinary feed group, FBG in the mice of high fat feed group increased significantly. However, there was no difference in terms of FBG between high fat feed group and model group. BUN, Cr and urine UmAlb in the mice from model group increased significantly compared with ordinary feed group and high fat feed group (P<0.01). Compared with ordinary feed group, the kidney weight (KW) and the kidney/body weight (KW/BW)of mice in model group increased,and the renal pathological lesions of mice in model group were obvious compared with ordinary feed group; The part of drug intervention:As compared with group A, FBQ BUN, SCr and urine UmAlb in the mice of group B increased significantly (P<0.01), the expression level of nephrin and podocin mRNA and protein were markedly decreased(P< 0.01). After intervention of ZGJTYS, the renal pathological lesions of mice in group C and group D were significantly reduced compared with group B, all biochemical indicators above in the mice of group C and D significantly decreased (P<0.01), the expression level of nephrin and podocin mRNA and protein were markedly increased(P<0.01) compared with group B. However, there was no statistical significance in terms of all indicators above between group C and group D (P>0.05)Conclusion This model established by high-fat diet feed and mono-nephrectomy is ideal. ZuoGui JiangTang YiShen decoction exerts reno-protective effect via reinstating nephrin and podocin expression to repair the damaged podocyte.

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