Experimental Study On The Effect Of The Decoction That Could Tonify Kidney And Invigorate The Blood Circulation,drive The Wind, On The MDR1 And P-gp170 In Rats With Adriamycin Nephropathy

Objective Through animal experiment,to observe the effect of Bushen Huoxue Qufeng decoction itself and(or)combining with glucocorticoid on MDR1 and glycoprotein P-gp170 in rats with adriamycin-induced nephropathy;Studying on Bushen Huoxue Qufeng therapy and experience in proteinuria induction,improving glucocorticoid sensitivity,inhanceing the treatment effection of nephrotic syndrome from the perspective of molecular biology mechanism.So as to provide an objective basis for traditional Chinese and combination of Chinese traditional and Western medicine for the treatment of refractory nephropathy.Methods Selecting healthy male SPF Spraque-Dawley rats,make sure every rat,s urine protein was negative,making 92 adriamycin nephropathy rats and 20 control group(A group),then rats were randomly divided into model group(group B),prednisone group(C group),Chinese medicine(group D),Chinese medicine prednisone group(group E),23 rats in each group.Traditional Chinese medicine group were treated with Bushen Huoxue Qufeng decoction,prednisone group was given the Prednisone Acetate Tablets solution,model group were given distilled water,the blank control group without intervention.The whole experimental time lasted for 8 weeks.At the end of the 4th week and 8th week,10 rats were randomly selected each group for observation of the treatment effect(24h urinary protein,renal function,blood lipid,blood coagulation),we used flow cytometry to measure the expression of P-gp170 in peripheral blood mononuclear cell membrane,and RT-PCR for peripheral blood mononuclear cells and renal tissue MDR1 m RNA,we collected kidney tissues for pathological check,and implyed immunohistochemistry for P-gp170 expression.Results(1)Rats in group B fur loose and pale,activity decreased,teeth broken,limbs and tail swelling,peritoneal effusion,meanwhile,the intervention group improve the situation,E group activities,diet increased significantly,but compared with D group appeared restless,increased death.(2)At the end of 8th week,the urinary protein was significantly lower in the prednisone group and goup E than that in the model group(P<0.05).The 24 h urinary protein ratio of the group E was lower than that of the prednisone group and goup D,all of their variation trend were accord with the curve(P> 0.05).(3)The lipoidemia: group A was lower than other groups(P<0.05),at the same time point.The group D was lower than that of group B?C(P<0.05).The TG: group A was lower than that of groups(P<0.05),at the same time.The TG level of group D was lowest when comparing with the B ? C ? E groups,especially with group C,the P<0.01.(4)The expression of MDR1 and P-gp170 in group B was significantly higher than that of group A(P <0.05).(5)The expression of MDR1 and P-gp170 in peripheral blood lymphocytes and renal tissue of prednisone group were sgnificantly higher than that of both blank group and model group(P <0.01).(6)The expression of P-gp170 in peripheral blood lymphocytes was lower than that in prednisone group(P <0.05).(7)In peripheral blood lymphocytes,the P-gp170 and MDR1 were positively correlated r = 0.466,p = 0.001.(8)P-gp170 in renal tissue was positively correlated with MDR1 m RNA r=0.291 p=0.291.(9)There was no significant difference about the expression of MDR1 and p-gp170 both in group D and group E,when compared with group B(P> 0.05).Conclusion(1)Bu Shen Huo Xue Qu Feng decoction can relieve the urine protein,control hyperlipidemia of rats with adriamycin nephropathy and reduce the pathological damage of kidney.(2)Glucocorticoid may induced the over expression of P-gp170,and the high level of P-gp170 may cause glucocorticoid resistance and reduce the effect of glucocorticoid in adriamycin nephropathy rats.(3)The level of P-gp170 in renal tissue and lymphocyte of rats with adriamycin nephropathy may affect the amount of urinary protein.(4)Bushen Huoxue Qufeng decoction to some degree,reduces the level of MDR1 m RNA and P-gp170 in rats with adriamycin induced nephropathy,or antagonizes their side effect,which may be one of the possible mechanisms to enhance the effect of the combination of glucocorticoid and reverse the drug resistance.