抗纤灵方通过PI3K/AKT/mTOR信号通路干预肾纤维化机制研究

Mechanism of Kangxianling Docotion in Intervening Renal Fibrosis Through PI3K / AKT / mTOR Signaling Pathway

目的:研究抗纤灵方对PI3K/AKT/mTOR信号通路的影响及抗肾纤维化作用机制。方法:将60只C57小鼠,随机分为假手术组10只和手术组50只,手术组行5/6肾切除术。术后2周,手术组随机分为模型组、抗纤灵低、中、高剂量组及雷帕霉素阳性药组,各组10只。假手术组给予0.5 mL生理盐水ig,抗纤灵方低、中、高剂量组分别给予0.5 mL抗纤灵药物ig(0.1,0.2,0.4 mg·kg-1),阳性药组给予0.5 mL雷帕霉素ig(0.016μg·kg-1),ig 12周后处死小鼠,在处死小鼠前1 d收集24 h尿液检测24 h蛋白定量,眼眶采血测血肌酐、尿素氮,取残肾采用HE观察肾脏组织形态改变,PCR法检测肾组织中PI3K/AKT/mTOR mRNA表达。结果:与假手术组比较,模型组24 h尿蛋白定量,血肌酐,尿素氮,PI3K/AKT/mTOR mRNA表达均显著升高(P<0.01),肾脏病理形态改变明显;与模型组比较,各治疗组24 h尿蛋白定量,血肌酐,尿素氮,PI3K/AKT/mTOR mRNA表达均下降(P<0.05),肾脏组织学形态改善。结论:抗纤灵方能降低小鼠24 h尿蛋白定量,改善肾功能,延缓肾纤维化发生;其机制可能与抑制PI3K/AKT/mTOR信号通路表达相关。

Objective： To study the effect of Kangxianling docotion on PI3K/AKT/mTOR signaling pathway and its mechanism against renal fibrosis. Method： Totally 60 C57 mice were randomly divided into sham group （n = 10） and operation group （n = 50） , the surgical group received 5/6 nephrectomy. After two weeks, the surgery group were randomly divided into model group, Kangxianling low, medium and high dose groups and rapamycin positive control group, with 10 in each group. The sham group received 0.5 mL of normal saline, ig, Kangxianling docotion low, medium and high dose groups were given 0.5 mL Kangxianling drug orally （0. 1 , 0.2, 0.4 mg·kg^-1） , rapamycin group was given 0.5 mL rapamycin gavage （0. 016 μg·kg^-1 ）. Mice were sacrificed after 12 weeks of gavage. The 24 h urine protein before the mice were sacrificed to detect 24 h protein quantification, the orbital blood was collected to test serum creatinine, blood urea nitrogen; the remnant kidney was collected to observe renal morphology by HE change; PCR was used to detect PI3K/AKT/mTOR mRNA expressions in kidney tissues. Result： Compared with sham operation group, model group showed significantly higher 24-hour urinary protein excretion, serum creatinine, blood urea nitrogen, PI3K/AKT/mTOR mRNA expressions （P 〈 0.01 ）, with significantly renal pathological changes. Compared with the model group, all treatment groups showed lower 24-hour urinary protein excretion, serum creatinine, blood urea nitrogen, PI3K/ AKT/mTOR mRNA expressions （P 〈 0.05） , with improvements in renal histology. Conclusion： Kangxianling can decrease 24 h urinary protein quantification in mice, improve renal function and delay the occurrence of renal fibrosis; its mechanism may be related to the inhibition of the expression pathway PI3K/AKT/mTOR signal.