孕酮对人孕晚期DSCs体外增殖及TNF-α、IL-6基因表达水平的影响  

Effects of progesterone on the proliferation and expressions of TNF-αand IL-6 genes of human decidual stromal cells in late pregnancy in vitro

作　　者：逯非凡 李媛 顾仲毅 李励 杨倩倩 徐畅 邓轶方[2] 管睿 Lu Feifan;Li Yuan;Gu Zhongyi;Li Li;Yang Qianqian;Xu Chang;Deng Yifang;Guan Rui(Department of Obstetrics and Gynecology,the First Affiliated Hospital,Naval Medical University,Shanghai 200433,China;Department of Pharmacology,Shanghai Institute of Pharmaceutical Industry,Shanghai 200437,China)

机构地区：[1]海军军医大学第一附属医院妇产科,上海200433 [2]上海医药工业研究院有限公司药理评价研究中心,上海200437

出　　处：《中华生殖与避孕杂志》2023年第7期718-722,共5页Chinese Journal of Reproduction and Contraception

基　　金：上海市自然科学基金(13ZR1408600);上海老龄化和妇儿健康研究专项(2020YJZX0216)。

摘　　要：目的研究孕激素对人孕晚期蜕膜基质细胞(decidual stromal cells,DSCs)形态、增殖以及分泌细胞因子的影响,从而探究孕激素在预防自发性早产中的作用机制。方法人孕晚期DSCs原代培养并鉴定,培养液中加入不同浓度的孕酮,分成10^(-6) mol/L组、10^(-5) mol/L组和10^(-4) mol/L组,对照组培养液不加孕酮。显微镜下观察各组DSCs细胞形态、测量细胞长/宽比值;四甲基偶氮唑盐比色(methyl thiazolyl tetrazolium,MTT)法测定细胞增殖活性;RT-qPCR检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)mRNA表达水平。结果①DSCs长/宽比值比较:孕酮10^(-4) mol/L组(5.87±0.19)、10^(-5) mol/L组(5.98±0.27)低于对照组(6.42±0.19),差异有统计学意义(P<0.001,P=0.002),孕酮10^(-4) mol/L组低于10^(-6) mol/L组(6.28±0.32),差异有统计学意义(P=0.005)。②细胞增殖活性比较:孕酮10^(-5) mol/L组(0.70±0.04)、10^(-4) mol/L组(0.78±0.04)分别高于对照组(0.59±0.05;P=0.027、P=0.002),10^(-4) mol/L组高于10^(-6) mol/L组(0.61±0.01,P=0.004)。③TNF-αmRNA表达比较:孕酮各浓度组均低于对照组(均P<0.001),10^(-5) mol/L组、10^(-4) mol/L组均低于10^(-6) mol/L组(均P<0.001);IL-6 mRNA表达比较:对照组、10^(-6) mol/L组、10^(-5) mol/L组和10^(-4) mol/L组中,IL-6 mRNA表达水平逐渐降低,组间两两比较,差异均有统计学意义(均P<0.001)。结论孕酮能使人孕晚期DSCs形态宽大饱满、促进细胞增殖、降低TNF-α及IL-6 mRNA表达水平,提示孕激素在预防自发性早产中起一定作用。Objective To study the effects of progesterone on the morphology,proliferation,and secretion of cytokines of human decidual stromal cells(DSCs)in late pregnancy,and to explore the mechanism of progesterone in preventing spontaneous preterm birth.Methods Human decidual stromal cells in late pregnancy were cultured and treated with different concentrations of progesterone(in the experimental groups,10^(-6) mol/L,10^(-5) mol/L and 10^(-4) mol/L progesterone was added to the culture medium respectively,and no progesterone was added to the culture medium of control group).The morphology of DSCs was observed under the microscope,the cell length/width ratio was measured,the proliferation was detected by methyl thiazolyl tetrazolium(MTT)method,and the expressions of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNA were detected by RT-qPCR.Results The length/width ratios of DSCs in progesterone 10^(-4) mol/L(5.87±0.19)and 10^(-5) mol/L(5.98±0.27)groups were lower than that in control group(6.42±0.19),the differences were statistically significant(P<0.001,P=0.002).The length/width ratio in the 10^(-4) mol/L group was lower than that in the 10^(-6) mol/L group(6.28±0.32,P=0.005).The proliferation of DSCs in the 10^(-5) mol/L(0.70±0.04)and 10^(-4) mol/L(0.78±0.04)groups was higher than that in control group(0.59±0.05;P=0.027,P=0.002),and proliferation of DSCs in 10^(-4) mol/L group was higher than that in 10^(-6) mol/L group(0.61±0.01,P=0.004).The expression of TNF-αmRNA in each progesterone group was lower than that in control group(all P<0.001)and the expression of TNF-αmRNA in the 10^(-5) mol/L and 10^(-4) mol/L groups was lower than that in the 10^(-6) mol/L group(all P<0.001).The expressions of IL-6 mRNA decreased gradually in control group,10^(-6) mol/L,10^(-5) mol/L and 10^(-4) mol/L groups,the differences were statistically significant(all P<0.001).Conclusion Progesterone can make the decidual stromal cells wider,promote proliferation,and decrease the expressions of TNF-αand IL-6 mRNA,which may

关 键 词：孕酮 细胞增殖 肿瘤坏死因子-α 白细胞介素-6 蜕膜基质细胞 

分 类 号：R714.21[医药卫生—妇产科学]