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目的:观察冬虫夏草主要有效成分——虫草素(cordycepin)对肾间质纤维化及其e IF2α/TGF-β/Smad信号通路的干预作用和机制。方法:将15只C57BL/6小鼠随机分为对照组、模型组和虫草素组,建立小鼠单侧输尿管结扎(unilateral ureteral obstruction,UUO)肾间质纤维化模型,虫草素组小鼠经皮下注射虫草素(5 mg·kg-1·d-1),同时,以生理盐水干预对照组和模型组,共5 d。实验结束后,取小鼠结扎侧的肾脏,观察肾间质纤维化病理特征,并测定肾组织I型胶原(collagen type I,Col I)和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)核酸表达(Northern blot);另外,在体外培养大鼠肾小管上皮细胞(NRK-52E细胞),在转化生长因子(transforming growth factor,TGF)-β联合或不联合虫草素干预后,观察I型胶原、IV型胶原(collagen type IV,Col IV)核酸表达的变化(Northern blot),并检测e IF2α、磷酸化e IF2α(phosphorylated e IF2α,p-e IF2α)以及Smad2/3、磷酸化Smad2/3(phosphorylated Smad2/3,p-Smad2/3)蛋白表达的变化(Western blot)。结果:虫草素在体内改善模型鼠肾间质纤维化,包括纤维化的病理特征以及I型胶原、α-SMA核酸的表达;虫草素在体外促进NRK-52E细胞p-e IF2α的高表达,抑制TGF-β诱导的p-Smad2/3以及I型、IV型胶原的表达水平。结论:虫草素在体内有改善肾间质纤维化的作用,它可能是通过诱导e IF2α磷酸化,抑制TGF-β/Smad信号通路中的关键信号分子——p-Smad2/3表达,减少肾组织胶原和α-SMA表达等途径而改善肾间质纤维化。
OBJECTIVE: To observe the intervention effect and mechanism of cordycepin, a main active component of Cordyceps, on renal interstitial fibrosis and eIF2α / TGF-β / Smad signaling pathway. Methods: Fifteen C57BL / 6 mice were randomly divided into control group, model group and cordycepin group. The model of unilateral ureteral obstruction (UUO) Cordycepin (5 mg · kg-1 · d-1) was injected subcutaneously, meanwhile, the rats in the control group and the model group were treated with saline for 5 days. At the end of the experiment, the kidneys from the ligation side of the mice were sacrificed and the pathological features of renal interstitial fibrosis were observed. The expressions of collagen type I (Col I) and α-smooth muscle actin α-SMA). In addition, rat renal tubular epithelial cells (NRK-52E cells) were cultured in vitro. After the intervention of transforming growth factor (TGF) -β or cordycepin The changes of the expression of collagen type IV (Col IV) and the expression of eIF2α, phosphorylated eIF2α and phosphatidylinfus, Smad2 / 3 (phosphorylated Smad2 / 3, p-Smad2 / 3) protein expression changes (Western blot). Results: Cordycepin improved renal interstitial fibrosis, including the pathological features of fibrosis and the expression of type I collagen and α-SMA in vivo. Cordycepin promoted the expression of pe IF2α in NRK-52E cells in vitro and inhibited the expression of TGF- β-induced p-Smad2 / 3 and type I, type IV collagen expression levels. CONCLUSION: Cordycepin can improve renal interstitial fibrosis in vivo. It may be through the induction of eIF2α phosphorylation, which inhibits the expression of p-Smad2 / 3, a key signaling molecule in TGF-β / Smad signaling pathway, Tissue collagen and α-SMA expression and other ways to improve renal interstitial fibrosis.