川芎嗪对单侧输尿管梗阻大鼠肾小管上皮细胞转分化的影响

Influence of ligustrazine on renal tubular epithelial mesenchymal transition in unilateral ureteral obstruction rats

目的观察川芎嗪对单侧输尿管梗阻大鼠(UUO)肾小管上皮细胞-间充质转分化(EMT)的影响及机制。方法 SD大鼠60只,随机分为A组(假手术)、B组(UUO)、C组[川芎嗪,40 mL/(kg·d),腹腔注射]、D组[厄贝沙坦,50 mg/(kg·d),灌胃],每组各15只。术后分别于第3、7和14天取左肾组织。荧光实时定量PCR检测分化抑制因子1(Id1)mRNA,免疫组化法测Id1、小窝蛋白1(Cav1)、转化生长因子-β1(TGFβ1)、钙粘蛋白(E-cad)、α-平滑肌肌动蛋白(α-SMA)、I型胶原表达及相关分析。结果与A组比较,B组大鼠各时间点Id1 mRNA和蛋白、Cav1、TGFβ1、α-SMA和I型胶原表达增多,E-cad表达逐渐减少;C组大鼠各时间点均反向调节上述因子,疗效与D组相似;UUO大鼠肾组织中Id1表达与Cav1、α-SMA、TGFβ1表达正相关,与E-cad表达呈负相关。结论 Id1及Cav1表达强弱与肾间质纤维化程度相关,川芎嗪能有效减轻UUO诱导的大鼠肾间质纤维化,可能与有效抑制Id1及Cav1表达,部分逆转EMT有关。

【Objective】To investigate influence and mechanism of ligustrazine on renal tubular epithelial mesenchymal transition. 【Methods】Sixty male, Sprague-Dauley（SD） rats, were randomly divided into the following 4groups： A group（sham group, n =15）, B group（UUO group, n =15）, C group（ligustrazine group, 40 mL/（kg·d）, intraperitoneal injection, n =15）, D group（irbesartan group, 50 mg/（kg·d）, by gavageand, n =15）. On the 3rd day, 7th day and 14 th day after operation, left kidney removed and the rats were sacrificed by exsanguinations after left kidney removed. The expression of Id1 mRNA in each groupsReal-time PCR was detected. The inhibitor of differentiation 1（Id1）, caveolin-1（Cav1）, TGFβ1, E-cadherin, α-smooth muscleactin（α-SMA） and collagenⅠ（ColⅠ）were detected by immunohistochemical staining, then correlation analysis was calculated. 【Results】 Compared with A group, the expression of Id1 at mRNA and protein, Cav1, TGFβ1, α-SMA and colⅠ were significantly increased in the B group（3 d, 7 d, 14 d） after the surgery（P 0.05, respectively）, E-cadherin was decreased. Treatment with Ligustrazine significantly abrogated such effects（P 0.05）; the therapeutic effect was similar to irbesartan. Positive areas of Id1 and expression of Cav1, α-SMA, TGFβ1 in renal tissues were significantly positive correlated, and Ecadherin protein expression was negatively correlated. 【Conclusion】The expression of Id1 and Cav1 reflects the level of renal interstitial fibrosis, Ligustrazine can effectively reduced the UUO-induced renal interstitial fibrosis,which may inhibit the expression of Id1 and Cav1, partially reversed EMT related.