| Objective: Diabetic nephropathy(diabetic nephropathy, DN) is referring to glomerular lesions caused by diabetes, the obvious pathological change that the extracellular matrix hyperplasia is a serious complication in diabetes developed to a certain extent. diabetic nephropathy model was established by feeding with high fat diet and given a low dose of STZ, Applying of Huayu Tongluo chinese trditionol medicine or irbesartan to the model, by observing the general conditions of rats, blood biochemical index, morphological changes of renal tissue, quantitative measurement of 24 h urinary protein, blood regular test, and the expression levelof connective tissue growth facter(CTGF), to study the Huayu Tongluo medicine role, and to explore its possible mechanisise for the prevension and control in DN.Methods: 70 healthy male SD rats, 10 were randomly divided into normal control group, 60 as model group by weight, the model group were fed with high sugar and high fat diet four weeks, then one-time given a low dose of injection of STZ 35 mg/kg, 72 hours after three consecutive caudal vein random blood glucose measurement ≥16.7mmol/L for building successful model, a total of 50 rats, were randomly divided into 4 groups: 14 as model group, 12 rats as irbesartan group, 12 rats as Huayu group, 12 as Tongluo group. treated groups were given corresponding drugs, in the 16 th at the week end, collecting 24 hour urine, blood sugar test paper measure fasting blood glucose(FBG); fasted for 12 hours, abdominal aortic blood serum separated, for blood biochemical detection in narcotism;And lay the kidney tissues for light microscopy, electron microscopy, immunohistochemistry and PCR.Results:1 Rat’s general statusNormal control group rats were generally in good conditions; Model group and treated group gradually eat more, drunk more, urinated more, became thinner and that were most significant in the model group. and also weight losing, color darking, spirit withering, performed most serious in model group, symptoms of rats in the treated group reduced somewhat light. 2 Results of water-intake, diet-intake and urine-outputCompared with control group, the water-intake, diet-intake and urine-output of rats increased significantly in the model group, irbesartan group and Huayu group Tongluo group(P < 0.05).Compared with model group, the water-intake decreased significantly in irbesartan group and Tongluo group(P < 0.05),the urine-output decreased significantly in irbesartan group and Tongluo group compared with Huayu group(P < 0.05); the water-intake, diet-intake were not statistically significant compared in irbesartan group and Tongluo group(P > 0.05). Compared with Huayu group, the water-intake diet-intake decreased significantly in irbesartan group and Tongluo group(P < 0.05) 3 The comparison of rats′ body weight and the kidney hypertrophy index( right kidney weight /body weight)Compared with control group, the body weight was significantly lighter in the model group and treatment groups(P <0.05), hypertrophy index in the model group and treatment groups were significantly higher(P <0.05); Compared with model group, the body weights were significantly lighter in treatment groups(P <0.05), compared within treatment groups, differences of the body weights and the kidney hypertrophy indexs were not statistically significant(P > 0.05). 4 The 24-hour urinary total protein(24h UTP)Compared with normal control group, model group the 24 h total urinary protein increased significantly(P < 0.05); Compared with model group, the treatment groups of 24 h urinary protein amounts decreased obviously(P < 0.05); the 24 h total urinary protein decreased significantly in irbesartan group and Tongluo groupcompared to Huayu group(P < 0.05). 5 The comparison of rats′ FBG, TC, TGCompared with normal control group, model group rats′ FBG, TC, TG increased significantly(P < 0.05); Compared with model group, the treatment group rats′ FBG, TC, TG decreased significantly(P< 0.05); Compared intreatment groups the TG TC decreased significantly in huayu and Tongluo groups, meantime TC TG decreased significantly in Tongluo group compared to huayu group(P < 0.05). 6 The comparison of rats′ ScrCompared with normal control group, the Scr increased significantly in the model group(P <0.05), compared with model group, the Scr decreased in the treatment groups(P <0.05); compared within treatment groups, the Scr was not statistically significant(P >0.05). 7 The observation of kidney pathological morphology 7.1 Light microscopy observationsThe Normal control group rats had normal kidney tissues structure;Model group rats′ renal glomerular became hypertrophy, renal capsule fissure shaped slit cavity change, glomerular basement membrane became diffuse thickening, and mesangial matrix became hyperplasia. Pathological changes in the Tongluo group rats were lighter than Huayu and irbesartan groups. 7. 2 Electronic microscope resultsNormal control group rats had normal kidney tissues structure;Model group rats renal glomerular basement membrane became thickening, increased mesangial matrix, widely fused podocyte foot processes, Pathological changes of the treatment group rats were lighter, comepared with the model group. each treatment group pathology changes significantly reduced; among the treatment groups, the Huayu group pathology performance was slightly heavier than the other treatment groups. 8 Immunohistochemical detection of renal tissue CTGFCTGF in rat kidney mainly expressed in renal tubules, renal interstitial, glomerular,especially in the mesangial matrix. The amount expression compared with normal control group, model group significantly increased(P < 0.05); Compared with model group, the expression quantity of treatment groups was obviously lower(P < 0.05);compared within treatment groups, the expression quantity of Huayu was slightly heavier than the other treatment groups.(P < 0.05). 9 The R-T PCR detection of renal cortical CTGFcompared with normal control group,The expression of renal cortical CTGF in model group was significantly higher(P < 0.05); Compared with model group, the expression quantity of treatment groups was obviously lower(P < 0.05); compared within treatment groups, the expression quantity of Huayu was slightly heavier than the other treatment groups(P < 0.05).Conclusions1 Huayutongluo Chinese medicine can significantly improve the general situation of DN rats, decrease the 24 h urine protein, reduce the pathological damage in DN, improve lipid metabolism disorder in diabetes, show kidney protection role.2 Huayutongluo Chinese traditional medicine can inhibit renal CTGF expression,show the similar protection role with irbesartan which may be associated with inhibition of cell proliferation.3 Tongluo medicine is superior to Huayu medicine in the protection role of DN.Especiely in the decrese of water-intake, diet-intake and urine-output, TC, 24 h UTP and pathological changes. |
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