| Objective: The experiment adopted gavage of bovine serum albumin(BSA+CCL4 subcutaneous injection),and caudal intravenous injection of LPS to develop IgA nephropathy rat model,and given Huoxue Tongluo Recipe treatment,to explore the IgA nephropathy model of experimental rat health,the24-hour urinary protein,blood biochemical indexes and renal pathological structure the change in rat kidney tissue,IL-13,IL-4 of the change,and serum CIC concentration to reveal the mechanisms of Huoxue Tongluo Qingre Lishi Recipe to IgA nephropathy.Methods:40 SD rats weighing 180-200 g were fed in a relatively stable environment for 1 week,then these rats were randomly allocated into four groups,including normal control group,model group,benazepril group and Chinese traditional medicine group.Secondly,making model for the four groups based on the improved IgA nephropathy rat model method proposed by Zhang and Li et al.In the next day,such rats were drenched with the bovine serum albumin(BSA)of 600mg/kg for 12 weeks.BSA of 600mg/kg was prepared through mixing the BSA and distilled water in 1:100.The injection way is subcutaneous injection with CCL4 of 0.1 ml and castor oil of 0.3 ml,once a week for 12 weeks;meanwhile,tail vein injection with LPS of 0.05 mg was implemented at weeks 13 and 14.Finally,the rats of the four groups were given the corresponding medicine by gavage at 1 week after the model was established successfully.At the end of the eighteenth removal of samples.In the weeks 8,12,14 and 18,these rates were fed in the metabolism cage with water but ambrosia.The 24-hour urine was collected to detect the 24-hour urinary protein quantitative.At the end of the eighteenth week,fasting for more than 12 hours,measuring body weight to the abdominal anesthetics,thebiochemical index and the serum CIC were detected by cotected the femoral artery blood,the expression of IL-13 and IL-4 was detected by real-time fluorescence quantitative PCR in renal issues.immunohistochemical determination of IL-13 expression.Results:1 The normal control group was sane,sleek,sensitive reaction,feeding and drinking were normal;the model group,the benazepril group,the Chinese traditional medicine group of rats were drenched,their coat gloss gradually became poor,listlessness,hair color significantly worse even hair removal,weight loss,slow reaction etc.After given the benazepril group and the Chinese traditional medicine group corresponding drug treatment,the two groups of rats of the mental and physical status were improved significantly,which compared with the model group.2 Compared with the normal control group,the model group 、 the benazepril group and the Chinese traditional medicine group’s 24-hour urinary protein quantitative increases significantly(P < 0.05),compared with the model group,the 24-hour urinary protein quantitative of the benazepril group and the Chinese traditional medicine group is lower significantly(P < 0.05),the benazepril group compared with the Chinese traditional medicine group,the 24-hour urinary protein quantitative is no significant change(P>0.05).3 The normal control group,the model group,the benazepril group,the Chinese traditional medicine group four groups ofCREA,BUN compared with no statistical significance(P>0.05).The rats in the normal control group were normal on serum lipid,the model group,the benazepril group,the Chinese traditional medicine group three groups compared with the normal control group,serum lipid increased significantly,with statistical significance(P<0.05),the benazepril group compared with the model group,serum lipid had no statistical significance(P>0.05);compared with the Chinese traditional medicine group,serum lipid had no statistical significance too(P>0.05)).4 Renal histopathological observationFrom the light microscope,the size of glomerulus of the normal controlgroup is normal,and the mesangium and matrix have no significant improvement.The size of glomerulus of the model group is normal,but the mesangium and matrix are thickening significantly.The mesangium of groups the Chinese traditional medicine group and he benazepril group are thicker than that of the normal control group.However,compared with the model group,the Chinese traditional medicine group and the benazepril group have a less pathological change degree.From the Ig A immunofluorescence : The normal control group glomerular mesangial had no immune complex deposition.In the model group we observed the fluorescent green immune complex deposition.the benazepril group,he Chinese traditional medicine group two groups have different degrees of immune complex deposition,but they are lighter than the model group.5 RT-PCR observationCompared with the normal control group,the IL-13 mRNA and IL-4mRNA expressions of the model group,the Chinese traditional medicine group and the benazepril group increase significantly(P < 0.05);Compared with the benazepril group and the model group,the IL-13 mRNA and IL-4mRNA expressions of he Chinese traditional medicine group decrease significantly(P < 0.05);Compared with the benazepril group,the IL-13 mRNA and IL-4 mRNA expressions of the Chinese traditional medicine group decrease significantly(P < 0.05).6 ELISA observationCompared with the normal control group,CIC concentration was significantly increased in the model group,the Chinese traditional medicine group and the benazepril group,which was statistical significance(P<0.05);the Chinese traditional medicine group and the benazepril group compared with the model group,CIC concentration decreased significantly,with statistical significance(P<0.05);the Chinese traditional medicine group compared to the benazepril group,the CIC concentration changed little,no statistical significance(P>0.05).Conclusion1 Huoxue Tongluo Qingre Lishi recipe can decrease the 24-hour urinary protein quantitative,and improve the renal pathological damage in rats with IgA nephropathy.This shows that Huoxue Tongluo Qingre Qushi recipe has certain therapeutic effect on IgA nephropathy,and it also has some protective effects on the kidney.2 Huoxue Tongluo Qingre Lishi recipe can regulate the expression of IL-13 and IL-4mRNA in renal tissue of IgA rats and decrease the concentration of the serum CIC. |
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