【摘要】 目的探讨清热益气化瘀通络方对糖尿病肾病(DN)大鼠足细胞表型转化即上皮-间质细胞转分化的作用。方法将40只健康雄性Wistar大鼠随机分为正常组、糖尿病肾病组、清热益气化瘀通络组、厄贝沙坦组,每组10只。除正常组大鼠外,其余组大鼠均采用腹腔注射链脲佐菌素方法制作DN模型。造模成功后,清热益气化瘀通络组和厄贝沙坦组给予相应药物灌胃,正常组和糖尿病肾病组给予等量生理盐水灌胃,均连续12周。灌胃结束后,检测各组大鼠血糖、血肌酐、尿素氮及24 h尿蛋白定量,应用实时定量RT-PCR法检测肾皮质中Wilms肿瘤抑制因子(WT1)、钙黏蛋白(P-Cadherin)、结蛋白(desmin)mRNA表达情况。结果实验结束时,各造模组的血糖水平均明显高于正常组(P均<0.05),但各造模组间血糖水平比较差异均无统计学意义(P均>0.05);各造模组的24 h尿蛋白定量也均明显高于正常组(P均<0.05),但厄贝沙坦组和清热益气化瘀通络组24 h尿蛋白定量均明显低于糖尿病肾病组(P均<0.05),且清热益气化瘀通络组明显低于厄贝沙坦组(P均<0.05)。WT1 mRNA表达(2-△Ct)及相对定量(2-△△Ct)结果显示从高到低依次为正常组、厄贝沙坦组、清热益气化瘀通络组、糖尿病肾病组,但4组间比较差异均无统计学意义(P均>0.05);正常组、厄贝沙坦组、清热益气化瘀通络组P-Cadherin mRNA表达(2-△Ct)及相对定量(2-△△Ct)均明显高于糖尿病肾病组(P均<0.05),desmin mRNA表达(2-△Ct)及相对定量(2-△△Ct)均明显低于糖尿病肾病组(P均<0.05),但3组间比较差异均无统计学意义(P均>0.05)。结论清热益气化瘀通络方可增加足细胞P-Cadherin表达而降低desmin表达,具有拮抗糖尿病肾病大鼠足细胞表型转化的作用。更多还原

【Abstract】 Objective It is to explore the effects of decoction for clearing heat,replenishing Qi,resolving stasis and unblocking collaterals on podocyte phenotypic transition( epithelial-to-mesenchymal transition,EMT) in diabetic nephropathy( DN) rats. Methods Forty healthy male Wistar rats were randomly divided into four groups: control group( group A),DN group( group B),group treated by clearing heat,replenishing Qi,resolving stasis and unblocking collaterals( group C) and group treated by Irbesartan group( group D),each group had 10 rats. All the rats except in group A were given streptozotoein by intraperitoneal injection to establish ND models. After model successfully established,group C and group D were given decoction for clearing heat,replenishing Qi,resolving stasis and unblocking collaterals and Irbesartan respectively by lavage,group A and group B were given the same dose of normal saline by lavage. All the groups were treated for12 weeks. At the end of experiment,blood sugar,SCr,BUN and 24-hour urinary protein quantity were measured,the mRNA expressions of Wilms tumor-inhibiting factor( WT1),P-cadherin and desmin were detected by RT-PCR. Results At the end of experiment,the level of blood sugar and 24 h urinary protein quantity in group B,group C and group D was higher than that in group A( P < 0. 05),but no significant difference in blood sugar was found among the latter 3 groups( P > 0. 05),the level of 24 h urinary protein quantity in group C and group D was lower than that in group B,the decrease in group C was more significant than that in group D( P < 0. 05). The results of the expression of WT1 mRNA( 2-△Ct)and relative quantity( 2-△△Ct) from high to low were group A,group D,group C and group B respectively,but no significant difference was found among the four groups( P > 0. 05). The expression of P-Cadherin mRNA( 2-△Ct) and relative quantity( 2-△△Ct) in group A,group D group C were significantly higher while the expression of desmin mRNA( 2-△Ct)and relative quantity( 2-△△Ct) were lower than those in group B( P < 0. 05),but no significant difference was found among group A,group C and group D( P > 0. 05). Conclusion The decoction for clearing heat,replenishing Qi,resolving stasis and unblocking collaterals had antagonistic effect on podocyte EMT through increasing the expression of P-cadherin and decreasing the expression of desmin in the rats with DN.更多还原