补肾固本方对大鼠骨质疏松模型中PI3K/AKT/mTOR信号通路表达的调控研究

目的观察补肾固本方对去卵巢骨质疏松(osteoporosis,OP)大鼠骨组织中PI3K/AKT/mTOR信号通路的影响,探讨补肾固本方干预OP的作用机制。方法将100只SD大鼠随机分为5组:正常组(C组)、模型组(M组)、补肾固本方组(B组)、补肾固本方+PI3k受体特异性阻断剂LY294002组(B+L组)、PI3k受体特异性阻断剂LY294002组(L组),除正常组外,其余4组建立去卵巢大鼠OP模型。药物连续干预12周后,采用ELISA方法检测血清中E2、BGP、ALP水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测各组股骨组织中LC3、Beclin1、caspase-9 mRNA的表达情况,蛋白质印迹法(Western blot)检测各组股骨组织中PI3K、AKT、mTOR蛋白的表达。结果补肾固本方可显著增高OP大鼠血清E2水平,降低BGP、ALP水平,显著上调LC3、Beclin1表达水平,降低caspase-9、PI3K、p-AKT、mTOR的表达;而上述变化能够被PI3k受体特异性阻断剂LY294002所阻断,且其差异具有统计学意义(P0.05)。结论补肾固本方减少去卵巢后大鼠OP的发生,其机制可能与抑制PI3K/AKT/mTOR信号通路及其下游基因蛋白表达有关。

Regulation of the expression of PI3K/AKT/mTOR signaling pathway by nourishing kidney and consolidating fundamental recipe in rat osteoporosis model

Objective To observe the effect of nourishing kidney and consolidating fundamental recipe on PI3K/AKT/mTOR signaling pathway in bone tissue of ovariectomized osteoporotic rats, and to explore the mechanism of nourishing kidney and consolidating fundamental recipe on osteoporosis (OP). Methods A total of 100 SD rats were randomly divided into five groups: normal group (group C), model group (group M), nourishing kidney and consolidating fundamental recipe group (group B), nourishing kidney and consolidating fundamental recipe + PI3k receptor specific blocker LY294002 group (group B + L), and PI3k receptor specific blocker LY294002 group (group L). Except in normal group, ovariectomized rat OP model was established in the other four groups. After 12 weeks of continuous drug intervention, the serum levels of E2, BGP, and ALP were detected using ELISA method. The expressions of LC3, Beclin1, and caspase-9 in femoral tissues were detected using real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The expressions of PI3K, AKT, and mTOR in femoral tissues were detected using Western blotting. Results Nourishing kidney and consolidating fundamental recipe significantly increased serum E2 level, reduced BGP and ALP levels, significantly increased LC3 and Beclin1 expression levels, and decreased caspase-9, PI3K, p-AKT, and mTOR expression in OP rats. The above changes could be blocked by PI3k receptor specific blocker LY294002, and the difference was statistically significant (P<0.05). Conclusion Nourishing kidney and consolidating fundamental recipe reduces the occurrence of OP in ovariectomized rats. Its mechanism may be related to the inhibition of PI3K/AKT/mTOR signaling pathway and downstream gene expressions.