摘要
本研究探讨了红景天多糖(rhodiola sachalinensis polysaccaride,RSP)对缺氧条件下猪睾丸间质细胞增殖和凋亡的影响及其机制.通过CCK-8法测定细胞活力,筛选出RSP的最适宜浓度和预处理时间;设正常组、低氧组、低氧+RSP组,流式细胞术检测细胞周期和凋亡率,RT-qPCR检测细胞增殖和凋亡相关基因(CCND1、CDK4、IL-6、TNF-a、Bcl-2、Bax、Caspase-3)mRNA 表达水平,Western blot 检测 Bax、Bcl-2、p21、JNK、p-JNK、p53 表达水平.低氧条件下,猪睾丸间质细胞活力下降,经0.012 5 mg·mL-1 RSP预处理18 h后,细胞活力有所提高.流式细胞术结果显示,RSP能缓解猪睾丸间质细胞低氧引起的细胞周期阻滞.RSP显著上调Bcl-2、CCND1、CDK4的表达(P＜0.05),下调 TNF-α、IL-6、Bax、Caspase-3、p21、p53、pJNK/JNK 的表达(P＜0.05).RSP 能保证低氧环境下猪睾丸间质细胞由S期进入G2期,促进细胞增殖,抑制细胞凋亡,推测RSP通过p53/p21和JNK通路调控细胞增殖和凋亡.
Abstract
This study investigates the effect and mechanism of rhodiola sachalinensis polysaccaride(RSP)on the proliferation and apoptosis of procine leydig cells under hypoxia conditions.The appropriate hypoxia injury time,optimal RSP concentration and the pre-treattime duration deter-mined by CCK-8 assay.The experiment was divided into 3 groups,normal group,hypoxia group,and hypoxia+RSP group,and cell cycle proliferation and apoptosis were detected by flow cytometry method,the cell proliferation and apoptosis-related genes(CCND1,CDK4,IL-6,TNF-a,Bcl-2,Bax and Caspase-3)mRNA expression levels was detect by RT-qPCR,Western blot was used to detect the expression of Bax,Bcl-2,p21,JNK,p-JNK and p53.The viability of porcine leydig cells was decreased under hypoxia,which could be alleviated after 18 hours of pretreatment with 0.012 5 mg·mL-1 RSP.The Flow cytometry results showed that RSP could alleviate cell cycle arrest induced by hypoxia in porcine leydig cells.RSP significantly up-regulated the expression of Bcl-2,CCND1,CDK4(P＜0.05),and down-regulated the expres-sion of TNF-α,IL-6,Bax,Caspase-3,and p21,p53,and pJNK/JNK(P＜0.05).RSP helps ensure the normal progression of leydig cells from S-phase to G2-phase under hypoxia conditions,promote cell proliferation and inhibit apoptosis.It is speculated that RSP regulates cell prolifera-tion and apoptosis through p53/p21 and JNK pathways.
基金项目
青海省自然基金应用基础研究(2022-ZJ-752)
维普
万方数据