隔药饼灸对慢性肾衰竭兔肾组织纤维化相关因子表达的影响

Effect of herbal cake-partitioned moxibustion on renal function and expression of renal connective tissue growth factor,integrin-linked kinase and bone morphogenetic protein-7 in chronic renal failure rabbits

目的:观察隔药饼灸对慢性肾衰竭(CRF)新西兰兔肾脏结缔组织生长因子(CTGF)、整合素连接激酶(ILK)、骨成型蛋白-7(BMP-7)表达的影响,探讨隔药饼灸干预CRF的可能机制。方法:清洁级雄性健康新西兰兔随机分为空白组、模型组、氯沙坦钾组、隔药饼灸组,每组6只。采用腺嘌呤灌胃制备CRF家兔模型。氯沙坦钾组予氯沙坦钾2.33 mg·kg-1·d-1灌胃治疗,隔药饼灸组予隔药饼灸"命门"、双侧"肾俞""脾俞"治疗,每天1次,每穴灸5壮,各组的治疗以12 d为1个疗程,每个疗程结束后休息2 d,连续治疗3个疗程。治疗结束后,采用全自动生化仪检测家兔血清肌酐(Scr)、血尿素氮(BUN)、24 h尿蛋白;Western blot法、实时荧光定量PCR法检测家兔肾组织CTGF、ILK、BMP-7蛋白及mRNA的表达水平。结果:与空白组比较,模型组Scr、BUN、24 h尿蛋白明显升高(P<0.01),提示造模成功。干预结束后,与模型组比较,隔药饼灸组Scr、BUN降低(P<0.01),氯沙坦钾组Scr、BUN及24 h尿蛋白均降低(P<0.01,P<0.05)。与空白组比较,模型组ILK、CTGF蛋白及mRNA相对表达量升高(P<0.01)、BMP-7蛋白及mRNA相对表达量降低(P<0.01);与模型组比较,氯沙坦钾组、隔药饼灸组ILK、CTGF蛋白及mRNA相对表达量均降低(P<0.01,P<0.05)、BMP-7蛋白及mRNA相对表达量升高(P<0.01)。与氯沙坦钾组比较,隔药饼灸组ILK蛋白表达量升高(P<0.01)、BMP-7蛋白表达量降低(P<0.01)。结论:隔药饼灸可通过抑制CRF兔肾组织中CTGF、ILK的表达,上调BMP-7的表达,改善细胞外基质的异常沉积,从而发挥对肾间质纤维化的治疗作用,这可能是隔药饼灸治疗CRF的机制之一。

Objective To observe the effect of herbal cake-partitioned moxibustion on renal function and expression of connective tissue growth factor(CTGF),integrin-linked kinase(ILK) and bone morphogenetic protein-7(BMP-7) in chronic renal failure(CRF) rabbits,so as to reveal its mechanisms underlying improvement of CRF.Methods Twenty-four male New Zealand rabbits were randomly divided into control,model,medication and herbal cake-partitioned moxibustion(moxibustion)groups(n=6 rabbits in each group).The CRF model was established by gavage of suspension of Adenine(150 mg·kg-1·d-1)for 21 days.Herbal cake-partitioned moxibustion was applied to"Mingmen"(GV4)and bilateral"Shenshu"(BL23),"Pishu"(BL20)and for 5 moxa-cones every time.Rabbits of the medication group was treated by gavage of Losartan Potassium(2.33 mg·kg-1·d-1).All the treatments were conducted once daily,12 times a course for consecutive 3 courses with a two-day rest after each course of treatment.Serum creatinine(Scr),blood urea nitrogen(BUN)and 24 h urine protein contents were detected by using an automatic biochemical analyzer.The expression levels of CTGF,ILK and BMP-7 proteins and mRNA in the kidney tissue were determined by Western blot and quantitative real time-PCR,separately.Results Following modeling,Scr and BUN and 24 h urine protein contents were significantly increased in the model group in comparison with the control group(P<0.01).After the intervention,Scr and BUN contents were all significantly decreased in both the moxibustion and medication groups relevant to the model group(P<0.01),suggesting an improvement of the renal function.Compared with the control group,the expression levels of ILK and CTGF mRNAs and proteins were obviously up-regulated(P<0.01),and those of BMP-7 mRNA and protein significantly decreased in the model group(P< 0.01).In comparison with the model group,the expression levels of ILK and CTGF mRNAs and proteins were significantly down-regulated in the two treatment groups(P<0.01,P<0.05),and those of BMP-7 mRNA and protein markedly increased in the two treatment groups(P<0.01).In comparison with the medication group,the expression level of ILK protein was significantly up-regulated(P<0.01)and BMP-7 protein was significantly down-regulated(P<0.01)in the moxibustion group.No significant differences were found between the medication and moxibustion groups in down-regulating the levels of Scr,BUN and 24 h urine protein and expression of ILK mRNA,CTGF mRNA and protein and BMP-7 mRNA(P>0.05).Conclusion Herbal cake-partitioned moxibustion can improve renal function in CRF rabbits,which may be related to its effects in suppressing the expression of ILK and CTGF,and in up-regulating the expression of BMP-7 in the kidney tissue.