| Objective: Diabetic nephropathy (DN) is the diabetic glomerular sclerosis, also is one of microvascular complications, and its incidence increased with the duration of diabetes. Patients died of diabetic renal failure increased gradually. The mechanism of DN is sophisticated, and modern medicine lacks of special effective treatment at present, so to find a way to prolong and treat DN is of great significance. In recent years, studies have found that herbs recipe has advantage for mulit-pathways and poly-targets treatment effection. The supplementing-qi and nourishing-yin resolving-masses and dredging-meridians(SNRD) recipe is our effective recipe based on the pathogenesis of DN and many years′clinical treatment. In cilical practice, good effection achived to treat DN. Privous study found it can improve clinical manifestions, decrease urinary protein, profect renal function,and prolong DN′s progress. On base of privous study, this study apply SNRD recipe to intervene DN rats model established by intraperitoneal injection of STZ, by observing their 24 hour urinary protein quantitative, renal pathology,and mcp-1 levels in renal tissues to confirm the action of SNRD recipe and probe its possible mechanism ,to provide an experimental basis and theoretical basis for its clinical application.Methods: Body weight 250g±10g, 95 healthy male SD rats were feed for adaptive for 1 week. Their urine glucose and protein were determined negative for urine collected through metabolic cage, then their left kidney was removed by way of incision bank.After 2 weeks, except blank group, remains injected intraperitoneally STZ 40mg/kg, tail vein blood was taken after 72hours. If Blood glucose was above 16.7mmol/l ,models were succeed made. Model rats were randomly divided into 5 equal groups: model group, irbesartan group,Traditional Chinese Medicine low dose group, middle dose group, high dose group. Model succeed 1 week, each group were given medicine at the dose of 20 times of adult dose qd. Irbesartan group was intragastric gavaged irbesartan 15mg/kg/d, Traditional Chinese Medicine low dose group,middle dose group,high dose group were given herbal 5.29g/kg/d,10.58g/kg/d,21.16g/kg/d, normal and model groups were gavaged equal quantitive saline every day for 12 weeks. Rats ate normal fodder, and ate or drunk freely with room temperature 18---20℃relative humidity 40%---70%, 12-hour alternating light. During the experiment, tail vein blood glucose was tested once a day. If blood glucose higher than or equal to 26mmol/l ,which may be life-threatening, proper amount of long-acting insulin(0.5---4u) injected subcutaneously in order to avoid ketosis as well reduce deaths. At the 12th weeks′end with treatment, all rats drunk freely but without food. Collected 24 hour urine, 24 hour urinary protein quantative was measured, then anesthesia the rats, after leaving blood sample,right kidney was took out, weighted, calculated hypertrophy indexes and collected the kidney tissue samples. Pathomorphology changes of kidney were observed under light microscope and electron microscope. RNA was extracted from each group renal cortex, reverse transcription polymerase chain reaction (RT-PCR)to detect the level of mcp-1mRNA.Results:1The body weight and hypertrophy index(kidney weight/body weight)of each groupModel group and each treatment group were lighter than normal group(P<0.05).There was no significant difference between model group and each treatment group for body weight (P>0.05). For hypertrophy index , model group and each treatment group were higher than normal group(P<0.05). There was no significant difference between model group and treatment groups, as well among each treated group(P>0.05). 2The 24hour urinary protein quantity of each group Model group and each treatment group increased significantly compared with normal group(P<0.05). Each treated group decreased significantly compared with model group(P<0.05). There was no significant difference among each treatment group(P>0.05).3 The pathomorphology changes of kidney3.1Light microscrope observationThe normal group showed that: the structure of glomerulus was intact,without glomerular hypertrophy, glomerular capillary mesangial, basement membrane and matrix did not show marked change. The model group showed that: glomerular hypertrophy, glomerular mesangial area widened, and mesangial matrix accumulate. Traditional Chinese Medicine group and irbesartan group showed different extent pathomorphology changes,but less than model group,i.e. glomerular hypertrophy slightly, small quantity mesangial cells and matrix accumulate .There was no significant difference among each group.3.2Observation under transmission electron microscopyThe normal group showed that: the glomerular basement memberance structural integrity, capillary endothelial cells and podocyte without anomalies. Model group showed that: glomerular basement memberance thickening, mesangial matrix increase, accompanied by extensive podocyte fusion. Traditional Chinese Medicine group and irbesartan group both showed different extent pathomorphology changes, but less than model group,i.e. glomerular capillary basement memberance thicken slightly, mesangial matrix accumulate ,accompanied by partial podocyte fusion.There was no significant difference among each group.4.test of mcp-1Model group and each treatment group increased significantly compared with normal group(P<0.05). Each treated group decreased significantly compared with model group(P<0.05). There was no significant difference among each treatment group(P>0.05). Conclusions:1. SNRD recipe can decrease urinary protein, improve DN rats′renal injury, and its dosage was no significant correlation; its role with western medicine irbesartan was considerable.2. SNRD recipe can inhibit the expression of mcp-1 in renal tissues. This may be one of the mechanisms for its prevention and treatment DN progress. |
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