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目的观察抗纤灵对5/6肾切除诱导的慢性肾脏纤维化小鼠磷酸化蛋白激酶B(p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(p-m TOR)及Akt,m TOR mRNA表达的影响,从Akt-m TOR信号通路角度探讨抗纤灵的作用机制。方法 C57BL/6J雄性小鼠采用5/6肾切除制备慢性肾脏纤维化模型。2周后根据肌酐水平将手术组小鼠分为模型组、雷帕霉素组和抗纤灵组,同时设立假手术组,每组10只。雷帕霉素组、抗纤灵组分别给予雷帕霉素(0.8μg/g)、抗纤灵(20 g/kg)每天0.5 ml灌胃治疗,模型组和假手术组灌胃等量蒸馏水。连续用药8周后处死小鼠,检测各组小鼠肌酐(Scr)、尿素氮(BUN)、24小时尿蛋白(24 h UP)含量;采用蛋白质印迹法(Western blotting)、RT-PCR检测肾脏组织中p-Akt、p-m TOR蛋白及Akt、m TOR mRNA的表达。结果与假手术组比较,模型组Scr、BUN、24 h UP、p-Akt、p-m TOR蛋白及Akt、m TOR mRNA显著升高(P<0.05,P<0.01)。与模型组比较,雷帕霉素组、抗纤灵组Scr、UN、24 h UP、p-Akt、p-m TOR蛋白及Akt,m TOR mRNA均有所降低(P<0.05,P<0.01)。雷帕霉素组与抗纤灵组比较,在降低Scr、24 h UP方面,抗纤灵组明显优于雷帕霉素组(P<0.05);在降低p-m TOR蛋白及m TOR mRNA方面,雷帕霉素优于抗纤灵组(P<0.05)。结论抗纤灵可能通过调节Akt-m TOR信号通路,对肾纤维化有一定的改善作用。
Objective To observe the effects of Kangxianling on 5/6 nephrectomy-induced chronic kidney fibrosis in mice phosphorylated protein kinase B (p-Akt), phosphorylated mammalian target of the rapamycin (pm TOR) and Akt, m TOR mRNA Expression of the impact of Akt-m TOR signaling pathway from the perspective of anti-fibrinol mechanism of action. Methods C57BL / 6J male mice were treated with 5/6 nephrectomy to prepare chronic kidney fibrosis model. After 2 weeks, the mice in the operation group were divided into model group, rapamycin group and Kangxianling group according to creatinine level. At the same time, sham-operation group was set up, with 10 rats in each group. Rapamycin group and Kangxianling group were administrated with rapamycin (0.8μg / g) and anti-fibrosis (20g / kg) 0.5mg per day respectively. The model group and sham-operated group were given the same amount of distilled water . The rats were sacrificed 8 weeks after the continuous administration, and the levels of Scr, BUN and 24 h urinary protein in each group were measured. The expression of protein was detected by Western blotting and RT-PCR Tissue expression of p-Akt, pm TOR protein and Akt, m TOR mRNA. Results Compared with sham operation group, the expression of Scr, BUN, 24 h UP, p-Akt, p-m TOR and Akt, m TOR mRNA in model group were significantly increased (P <0.05, P <0.01). Compared with model group, Scr, UN, 24 h UP, p-Akt, p-m TOR protein and Akt, m TOR mRNA in rapamycin group and anti-fibronectin group were decreased (P <0.05, P <0.01). In the rapamycin group and Kangxianling group, Kangxianling group was significantly better than rapamycin group in reducing Scr, 24 h UP (P <0.05); in reducing pm TOR protein and m TOR mRNA, Rapamycin was superior to Kangxianling group (P <0.05). Conclusion Kangxianling may improve renal fibrosis by regulating Akt-m TOR signaling pathway.