本研究观察三七对阿霉素诱导的肾纤维化血瘀证大鼠模型磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)信号通路及线粒体能量代谢的影响,探讨三七对肾脏的保护作用机制。将30只阿霉素诱导的雄性大鼠随机分为模型,低,medium-,和大剂量三七,阳性对照组(n=6)。选用6只干净SD雄性年夜鼠进入正常组。正常组和模型组给予生理盐水,和其他组相应的药物。治疗8周后,肾功能,肾脏病理学,三磷酸腺苷(ATP)水平,Na~+-K~+-ATP酶和Ca~(2+)-Mg~(2+)-ATP酶活性,和ATP5B的蛋白质水平,mTORC1,70kDa核糖体蛋白S6激酶(P70S6K),P85,Akt,p-Akt,测定肾组织中含有SH2的肌醇磷酸酶(SHIP2)。与正常组相比,模型组血尿素氮(BUN)和血肌酐(SCr)水平升高(P<0.01)。与模型组相比,三七和阳性对照降低了BUN和SCr的水平,中、高剂量诺参组及阳性对照组差异有统计学意义(P<0.05)。与模型组相比,三七和阳性对照减轻了肾组织的病理变化,如空泡和纤维瘤的变化,肾小球萎缩,肾小管囊性扩张,大量的炎症细胞浸润.与正常组相比,模型组大鼠肾组织线粒体ATP含量、Na~+-K~+-ATP酶和Ca~(2+)-Mg~(2+)-ATP酶活性降低(P<0.05),中、高剂量三七和阳性对照可缓解这种下降(P<0.05)。与模型组相比,中、高剂量三七和阳性对照上调ATP5B和SHIP2的蛋白水平,下调mTORC1、P70S6K的蛋白水平,P85,Akt,和p-Akt(P<0.05或P<0.01或P<0.001)。三七可能通过抑制PI3K/Akt/mTORC1通路的激活恢复线粒体能量代谢而发挥抗纤维化作用,从而保护肾脏。
This study observed the effects of Notoginseng Radix et Rhizoma on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin complex 1(mTORC1) signaling pathway and mitochondrial energy metabolism in the rat model of adriamycin-induced renal fibrosis with blood stasis syndrome to explore the mechanism of Notoginseng Radix et Rhizoma in protecting the kidney. Thirty male rats with adriamycin-induced renal fibrosis were randomized into model, low-, medium-, and high-dose Notoginseng Radix et Rhizoma, and positive control groups(n=6). Six clean SD male rats were selected into the normal group. The normal group and model group were administrated with normal saline, and other groups with corresponding drugs. After 8 weeks of treatment, the renal function, renal pathology, adenosine triphosphate(ATP) levels, Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities, and the protein levels of ATP5B, mTORC1, 70 kDa ribosomal protein S6 kinase(P70S6K), P85, Akt, p-Akt, and SH2-containing inositol phosphatase(SHIP2) in the renal tissue were determined. Compared with the normal group, the model group showed elevated levels of blood urea nitrogen(BUN) and serum creatinine(SCr)(P<0.01). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control lowered the levels of BUN and SCr, which were significant in the medium-and high-dose Noto-ginseng Radix et Rhizoma groups and the positive control group(P<0.05). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control alleviated the pathological changes in the renal tissue, such as vacuolar and fibroid changes, glomerulus atrophy, cystic expansion of renal tubules, and massive infiltration of inflammatory cells. Compared with the normal group, the model group showed decreased mitochondrial ATP content and Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities in the renal tissue(P<0.05), and medium-and high-dose Notoginseng Radix et Rhizoma and positive control mitigated such decreases(P<0.05). Compared with the model group, medium-and high-dose Notoginseng Radix et Rhizoma and the positive control up-regulated the protein levels of ATP5B and SHIP2 and down-regulated the protein levels of mTORC1, P70S6K, P85, Akt, and p-Akt(P<0.05 or P<0.01 or P<0.001). Notoginseng Radix et Rhizoma may exert an anti-fibrosis effect by inhibiting the activation of the PI3K/Akt/mTORC1 pathway to restore mitochondrial energy metabolism, thus protecting the kidney.