祛风通络方对肾小球系膜细胞间隙连接蛋白36及基因表达的影响

吴喜利; 叶冰玉; 安鹏等

祛风通络方对肾小球系膜细胞间隙连接蛋白36及基因表达的影响

Effect of Qufeng Tongluo Recipe on the Expressions of Connexin 36 Protein and Gene in Rat Mesangial Cells

摘要

摘要: 目的研究间隙连接蛋白36(Cx36)及其基因在肾小球系膜细胞（MCs）的表达，观察祛风通络方的干预作用，从而进一步探讨祛风通络方抑制肾小球MCs增殖的作用机制。方法应用血清药理学方法，制备贝那普利药物血清和祛风通络药物血清，体外培养肾小球MCs，并将其分为正常对照、脂多糖(LPS)、贝那普利治疗和祛风通络方治疗组。分别采用激光扫描共聚焦显微镜、Western blot、免疫组化及实时定量PCR检测各组肾小球系膜细胞Cx36蛋白及基因的表达变化。结果与对照组比较，脂多糖组肾小球MCs Cx36表达明显增强（P<0.01）；与脂多糖组比较，两治疗组Cx36表达均减弱（P<0.01或P<0.05）。各组肾小球MCs Cx36基因表达与蛋白表达相似。结论 Cx36及基因在脂多糖诱导的肾小球MCs中有明显表达，祛风通络方抑制肾小球系膜细胞增殖的作用可能与其调节Cx36及基因表达有关。

Abstract: 【Abstract】 Objective To determine the effect of Qufeng Tongluo Recipe (QFTLR) on the expressions of connexin 36 (Cx36) protein and gene in rat mesangial cells (MCs) and the proliferation of the MCs. Methods Serum samples containing Benazepril (Bena) and QFTLR were prepared in line with serum pharmacology methodology. The MCs culturedin vitro were divided into normal control and Lipopolysaccharide (LPS), Bena and QFTLR treated groups. The expressions of Cx36 protein and gene were detected by laser scanning confocal microscope (LSCM), Western blot, immunohistochemical assay and quantitative real time polymerase chain reaction (QRT-PCR) respectively. Results Compared with the control, higher level of Cx36 protein expression was found in the MCs than treated with LPS (P<0.01). Both Bena and QFTLR lowered the level of Cx36 protein expression in the MCs treated with LPS significantly (P<0.01 or P<0.05). Similar results were found with the expression of Cx36mRNA. Conclusion QFTLR inhibits the proliferation of rat MCs, possibly through down-regulating the expressions of Cx36 protein and gene.

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