基于玄府理论的固本通络方对IgA肾病大鼠Podocin mRNA和α-actinin-4 mRNA表达的影响

Effects of Guben Tongluo Formula on Expressions of Podocin mRNA and α-actinin-4 mRNA in Rats with IgA Nephropathy Based on Sweat Pore Theory

目的:探析基于玄府理论的固本通络方对IgA肾病大鼠Podocin mRNA和α-actinin-4 mRNA表达的调节及对IgA肾病蛋白尿的治疗作用。方法:将80只SD大鼠随机分为正常组、模型组、中药组和科素亚组,采用牛血清清蛋白+四氯化碳+蓖麻油+脂多糖的方法建立IgA肾病大鼠模型。分别于造模结束后、治疗2周、治疗4周后检测各组大鼠24 h尿蛋白定量、血肌酐、尿素氮水平。治疗2周和4周时,采用光镜、免疫荧光法观察肾小球形态学变化;PCR法检测Prodocin、α-actinin-4 mRNA表达量。结果:治疗前,正常组24 h尿蛋白定量明显少于模型组,差异具有统计学意义(P<0.05)。模型组、中药组及科素亚组肾脏病理示肾小球系膜增生,基底膜增厚,系膜区IgA荧光强度极高。经治疗,与模型组比较,中药组及科素亚组在治疗2周、4周后24 h尿蛋白定量明显下降,差异具有统计学意义(P<0.05);中药组在治疗2周、治疗4周后与科素亚组比较24 h尿蛋白定量均减少,差异具有统计学意义(P<0.05)。与模型组比较,治疗后中药组和科素亚组Podocin mRNA含量明显升高,α-actinin-4 mRNA含量均明显降低,差异具有统计学意义(P<0.05);治疗4周与治疗2周比较,科素亚组Podocin含量明显升高,中药组α-actinin-4 mRNA含量均明显降低,差异具有统计学意义(P<0.01);治疗4周后中药组α-actinin-4 mRNA含量均明显低于科素亚组,差异具有统计学意义(P<0.05)。中药组及科素亚组肾小球病变较治疗前损伤程度轻,荧光强度明显减低。结论:固本通络方可以减轻IgA肾病大鼠蛋白尿与调节足细胞Podocin和α-actinin-4表达有关。

Objective:To explore the effects of the Guben Tongluo Formula on the expression of Podocin mRNA and alpha-actinin-4 mRNA in IgA nephropat hy rats and the treatment of urinary protein in IgA nephropathy. Methods:Eighty SD rats were randomly divided into four goups:control group,model group,herb group and Cozaar group,with the method of Bovine serum albumin + carbon tetrachloride + castor oil + lipopolysaccharide establishing rat model of IgA nephropathy. The 24-hour urine protein quantitation,serum creatinine and urea nitrogen levels of each group were measured after the end of modeling,2 weeks and 4 weeks. At the end of 2 nd and 4 th week,the morphological changes of glomeruli were observed by light microscopy and immunofluorescence and PCR was uesd respectively to examine the mRNA expression of Podocin and α-actinin-4. Results:Before treatment,the 24-hour urine protein in the normal group was significantly less than that in the model group,which was statistically significant(P< 0.05). Renal pathology of the model group,Chinese medicine group and Kosuya group showed mesangial hyperplasia,thickening of the basement membrane,and high fluorescence intensity of IgA in the mesangial area. After treatment,at the end of 2 nd and 4 th week,the urine protein quantitation of the Chinese medicine group and Kosuya group was significantly lower than that in the model group,with statistical significance(P< 0.05). At the end of 2 nd and 4 th week,compared with the Cozaar group,the urinary protein quantitation of the Chinese medicine group was reduced and there was a statisti cal significance(P< 0.05). Compared with the model group,the content of Podocin mRNA of the Chinese medicine group and Kosuya group was significantly increased,and the content of α-actinin-4 mRNA was significantly decreased after the treatment(P< 0.05). At the end of 4 th week,compared with the results of treatment for 2 nd weeks,the content of Podocin in the Kossuya group was significantly increased,and the content of α-actinin-4 mRNA in the Chinese medicine group was significantly decreased(P< 0.01). The content of α-actinin-4 mRNA in the Chinese medicine group was significantly lower than that in the Kossuya group at the end of 4 th week(P< 0.05). The glomerular lesions in the Chinese medicine group and Kosuya group were lighter than before treatment,and the fluorescence intensity was significantly reduced. Conclusion:Guben TongLuo Formula can alleviate proteinuria in rats with IgA nephropathy,it is associated with the regulation of Podocin and α-actinin-4 expression in podocytes.