血必净通过激活PI3K/Akt/mTOR信号通路减轻大鼠睾丸缺血再灌注损伤

Xuebijing relieves testicular ischemia/reperfusion injury in rats by activating PI3K/Akt/mTOR signaling pathway

目的:探究血必净对缺血再灌注(I/R)损伤大鼠睾丸的保护作用及其相关机制。方法:45只雄性SD大鼠随机分为对照组、模型组、血必净低剂量组、血必净高剂量组和地塞米松组(均n=9);除对照组大鼠外,其它各组大鼠构建睾丸扭转复位模型,术后低、高剂量组及地塞米松组大鼠分别腹腔注射0.5和2 mL·kg^-1·d^-1血必净及0.5 mL·kg^-1·d^-1地塞米松。用药第3、7和14天取各组大鼠左侧睾丸,采用HE染色观察各组大鼠睾丸组织病理学改变;生化检测各组大鼠睾丸组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、内皮素1(ET-1)和一氧化氮(NO)水平,Western blot检测各组大鼠睾丸组织中细胞周期相关蛋白、细胞凋亡相关蛋白以及PI3K/Akt/mTOR信号通路相关蛋白的水平。结果:血必净可显著减轻I/R大鼠睾丸损伤,显著升高I/R大鼠睾丸组织中SOD活性,降低MDA、ET-1和NO的含量,抑制I/R损伤组织中的氧化应激,介导细胞周期和细胞凋亡相关因子的表达,并显著升高I/R大鼠睾丸中p-PI3K、p-AKT、p-mTOR和p-S6K的蛋白水平,其作用效果具有时间依赖性和剂量依赖性。结论:血必净通过介导细胞周期和细胞凋亡相关蛋白的表达,抑制氧化应激反应,激活PI3K/Akt/mTOR信号通路来减轻大鼠睾丸I/R损伤,且其作用效果呈剂量依赖性和时间依赖性。

AIM:To investigate the effect of Xuebijing on testicular ischemia/reperfusion(I/R)injury in rats and its related mechanisms.METHODS:Male Sprague-Dawley rats(n=45)were randomly divided into control group,I/R group,low-dose Xuebijing group,high-dose Xuebijing group and dexamethasone group(n=9 in each group).Except for the rats in control group,the rats in other groups underwent testicular torsion,and after the operation,the rats were treated with 0.5 mL·kg^-1·d^-1 Xuebijing,2 mL·kg^-1·d^-1 Xuebijing and 0.5 mL·kg^-1·d^-1 dexamethasone in low-dose Xuebijing group,high-dose Xuebijing group and dexamethasone group,respectively.On the 3rd,7th,and 14th days after treatment,the left testis in the rats of each group was taken.The histopathological changes of the testis were observed by hematoxylin-eosin staining.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),endothelin-1(ET-1)and nitric oxide(NO)in the testicular tissue were detected by biochemical methods.The protein levels of cell cycle-related molecules,apoptosis-related proteins and PI3K/Akt/mTOR signaling pathway-related proteins were determined by Western blot.RESULTS:Xuebijing significantly attenuated the testicular damage in I/R rats,significantly increased the activity of SOD in the testis of I/R rats,reduced the content of MDA,ET-1 and NO,inhibited oxidative stress in I/R-injured tissues,mediated the protein expression of cell cycle-related factors and apoptosis-related factors,and significantly increased the protein levels of p-PI3K,p-AKT,p-mTOR and p-S6K in the testis of I/R rats(P<0.05).These effects were time-dependent and dose-dependent.CONCLUSION:Xuebijing reduces testicular I/R injury of rats by mediating the expression of cell cycle-related and apoptosis-related proteins and activating PI3K/Akt/mTOR signaling pathway in dose-dependent and time-dependent manners.