益母缩宫颗粒含药血清对人早孕蜕膜腺基质细胞凋亡相关蛋白表达的影响  

Effect of Yimu Suogong Granules containing serum on the expression of apoptosis related proteins in human decidual gland stromal cells during early pregnancy

作　　者：陈艳 毛惠 赵雅 CHEN Yan;MAO Hui;ZHAO Ya(College of Integrated Chinese and Western Medicine,Southwest Medical University,Sichuan Province,Luzhou646000,China)

机构地区：[1]西南医科大学中西医结合学院,四川泸州646000

出　　处：《中国当代医药》2023年第14期20-26,共7页China Modern Medicine

摘　　要：目的研究益母缩宫颗粒(YMSGG)含药血清对人早孕蜕膜腺基质细胞增殖和凋亡的影响,同时探讨其可能的作用机制。方法制备YMSGG含药血清和空白血清,体外培养人早孕蜕膜腺基质细胞,采用CCK-8法筛选YMSGG含药血清最佳作用浓度和时间用于后续实验。将人早孕蜕膜腺基质细胞分为YMSGG含药血清低、中、高剂量组和空白血清低、中、高剂量组,利用流式细胞术检测YMSGG含药血清对人早孕蜕膜腺基质细胞凋亡的影响;应用实时荧光定量PCR(rt-PCR)法检测B淋巴细胞瘤-2(Bcl-2)、Bax、Caspase-3、Caspase-8、Caspase-9,Fas、Fas配体(Fasl)mRNA表达情况;通过Western blot检测人早孕蜕膜腺基质细胞中凋亡相关蛋白Bcl-2、Bax、Caspase-3、Caspase-8、Caspase-9、Fas、Fasl蛋白表达水平。结果干预12 h,各浓度含药血清组的OD值与空白血清组比较,差异无统计学意义(P>0.05);干预24、48 h,各浓度含药血清组的OD值高于空白血清组,差异有统计学意义(P<0.01);且抑制率随浓度的升高而升高,但48 h药物浓度与抑制率线性关系较24 h好,因此选择48 h作为最佳作用浓度时间。根据48 h药物浓度与抑制率曲线图,计算出IC50为0.155,故选择5%、10%、15%作为后续实验低中高浓度组。5%含药血清凋亡率与5%空白血清凋亡率差异无统计学意义(P>0.05);10%、15%含药血清凋亡率分别高于10%、15%空白血清凋亡率,差异有统计学意义(P<0.01);且随浓度升高而升高。5%、10%、15%YMSGG含药血清组Bcl-2 mRNA的表达分别低于5%、10%、15%空白血清组,差异有统计学意义(P<0.05);Bax、Caspase-3、Caspase-8、Caspase-9、Fas、FasL mRNA的表达分别高于5%、10%、15%空白血清组,差异有统计学意义(P<0.05)。随着YMSGG含药血清浓度的增加,人早孕蜕膜腺基质细胞表达Bax、Caspase-3、Caspase-8、Caspase-9、Fas、FasL呈一定的浓度依赖性。5%、10%、15%YMSGG含药血清组的Bcl-2蛋白表达分别低于5%、10%、15%空Objective To study the effect of Yimu Suogong Granules(YMSGG)containing serum on the proliferation and apoptosis of human decidual gland stromal cells in early pregnancy,and to explore its possible mechanism.Methods Preparation of drug containing serum and blank serum of YMSGG.Culture of human decidual gland stromal cells in early pregnancy in vitro.Cell counting kit(CCK-8)method was used to screen the best concentration and time of action of YMSGG-containing serum for subsequent experiments.Human decidual gland stromal cells in early pregnancy were divided into low,medium and high dose groups of YMSGG-containing serum and low,medium and high dose groups of blank serum.The effect of YMSGG-containing serum on apoptosis of human decidual gland stromal cells in early pregnancy was detected by flow cytometry.The expression of B lymphoblastoma-2(Bcl-2),Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fas ligand(Fasl)mRNA were detected by real-time PCR(rt-PCR).The expression levels of apoptosis related proteins Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9,Fas and Fasl in decidual gland stromal cells of early pregnancy were detected by Western blot.Results After 12 hours of intervention,there was no significant difference in OD value between the drug containing serum group and the blank serum group(P>0.05).After 24 and 48 hours of intervention,the OD value of drug containing serum group was significantly higher than that of blank serum group(P<0.01).The inhibition rate increased with the increase of concentration,but the linear relationship between drug concentration and inhibition rate at 48 h was better than that at 24 h.Therefore,48 h was chosen as the best concentration time;According to the curve of 48 h drug concentration and inhibition rate,IC50 was calculated to be 0.155,so 5%,10%and 15%were selected as the low,medium and high concentration groups for subsequent experiments.There was no significant difference between the apoptosis rate of 5%medicated serum and that of 5%blank serum(P>0.05).The apoptotic rates of 10%and 15%d

关 键 词：益母缩宫颗粒 细胞凋亡 B淋巴细胞瘤-2 Bax 死亡因子 FAS配体 蜕膜残留 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]