孕酮和脂多糖对人羊膜细胞β-防御素-4(HBD-4)表达的影响
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摘要
研究了孕酮(P4)和脂多糖(LPS)对人羊膜细胞内β-防御素-1(HBD-1) mRNA和β-防御素-4(HBD-4) mRNA表达的影响.使用10~(-9)M孕酮、0.5μg/mL脂多糖处理无血清羊膜细胞4 h、8 h、12 h、24 h,各时间点设无孕酮、无脂多糖添加的对照组(Control),然后使用10~(-9)M孕酮+0.5μg/mL脂多糖(P4+LPS)处理无血清羊膜细胞4 h、8 h、12 h、24 h,各时间点设0.5μg/mL脂多糖处理组(LPS)为对照组,并利用RT-qPCR技术检测各处理组中羊膜细胞内HBD-1、HBD-4 mRNA的表达变化.与对照组比较,孕酮和脂多糖对羊膜细胞HBD-1 mRNA的表达均无显著性影响.脂多糖可上调人羊膜细胞中HBD-4 mRNA的表达(P<0.05),而只添加孕酮对羊膜细胞中HBD-4 mRNA的表达量无显著影响,但在孕酮与脂多糖共同诱导下可上调人羊膜细胞中HBD-4 mRNA的表达(P<0.05).孕酮和脂多糖对羊膜细胞内HBD-1的表达无显著调节作用.羊膜细胞分泌的HBD-4是妊娠期羊膜腔天然免疫的重要组成部分,羊膜及羊膜腔感染时缺少孕激素将会影响羊膜细胞中HBD-4的表达,从而易引起抗菌能力降低而危及母亲和胎儿健康.
The RT-qPCR was used to investigate the effects of progesterone(P4) and lipopolysaccharide(LPS) on the expression of β-defensin-1(HBD-1) mRNA and β-defensin-4(HBD-4) mRNA in human amniotic cells in this research.To determine the expression of the two mRNA,Serum-free cultured amniotic cells were treated with(1) 10~(-9)M progesterone;(2) 0.5 μg/mL LPS;(3) 10~(-9)M progesterone+0.5 μg/mL LPS,for 4 h,8 h,12 h and 24 h,and progesterone-free and LPS-free as control for(1) and(2),0.5 μg/mL LPS for(3).The following results were obtained:(1) progesterone and LPS had no significant effect on the expression of HBD-1 mRNA;(2) LPS up-regulated the expression of HBD-4 mRNA(P<0.05);(3) progesterone had no significant effect on the expression of HBD-4 mRNA;(4) progesterone increased the expression of HBD-4 mRNA under the circumstances with LPS(P<0.05).HBD-4 secreted by amnion cells is an important component of the natural immunity of the amniotic cavity during pregnancy.The lack of progesterone will affect the expression of HBD-4 in amniotic cells,which may lead to the decrease of antibacterial ability and hence to endanger the health of mother and fetus,when the amniotic membrane and amniotic cavity are infected.
The RT-qPCR was used to investigate the effects of progesterone(P4) and lipopolysaccharide(LPS) on the expression of β-defensin-1(HBD-1) mRNA and β-defensin-4(HBD-4) mRNA in human amniotic cells in this research.To determine the expression of the two mRNA,Serum-free cultured amniotic cells were treated with(1) 10~(-9)M progesterone;(2) 0.5 μg/mL LPS;(3) 10~(-9)M progesterone+0.5 μg/mL LPS,for 4 h,8 h,12 h and 24 h,and progesterone-free and LPS-free as control for(1) and(2),0.5 μg/mL LPS for(3).The following results were obtained:(1) progesterone and LPS had no significant effect on the expression of HBD-1 mRNA;(2) LPS up-regulated the expression of HBD-4 mRNA(P<0.05);(3) progesterone had no significant effect on the expression of HBD-4 mRNA;(4) progesterone increased the expression of HBD-4 mRNA under the circumstances with LPS(P<0.05).HBD-4 secreted by amnion cells is an important component of the natural immunity of the amniotic cavity during pregnancy.The lack of progesterone will affect the expression of HBD-4 in amniotic cells,which may lead to the decrease of antibacterial ability and hence to endanger the health of mother and fetus,when the amniotic membrane and amniotic cavity are infected.
引文
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[2] Foerster K,He W,Manuel J,et al. LPS-induced occult loss in mice requires FGL2 [J]. Am J Reprod Immunol,2007(58):524-529.
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[4] King A E,Kelly R W,Sallenave J M,et al. Innate immune defences in the human uterus during pregnancy [J]. Placenta,2007,28(11-12):1099-1106.
[5] Butts C L,Bowers E,Horn J C,et al. Inhibitory effects of progesterone differ in dendritic cells from female and male rodents [J]. Gend Med,2008(5):434-447.
[6] Wira C R,Rodriguez-Garcia M,Patel M V. The role of sex hormones in immune protection of the female reproductive tract [J]. Nat Rev Immunol,2015,15(4):217-230.
[7] Han J H,Kim M S,Lee M Y,et al. Modulation of human beta-defensin-2 expression by 17beta-estradiol and progesterone in vaginal epithelial cell [J]. Cytokine,2010,49(2):209-214.
[8] Castro-Leyva V,Zaga-Clavellina V,Espejel-Nu,et al. Decidualization mediated by steroid hormones modulates the innate immunity in response to group B streptococcal infection in vitro [J]. Gynecol Obstet Invest,2017,82(6):592-600.
[9] Szekeres-Bartho. Immunological relationship between the mother and the fetus [J]. Int Rev Immunol,2002,21(6):471-495.
[10] Koga K,Mor G. Expression and function of toll-like receptors at the maternal-fetal interface [J]. Reprod Sci,2008,15(3):231-242.
[11] Radtke A L,O’Riordan M X. Intracellular innate resistance to bacterial pathogens [J]. Cell Microbiol,2006,8(11):1720-1729.
[12] Seo S J,Ahn S W,Hong C K,et al. Expression of β-defensins in human keratinocyte cell lines [J]. J Dermatological Sci,2001,27(3):183-191.
[13] King A E,Fleming D C,Critchley H O,et al. Differential expression of the natural antimicrobials,beta-defensins 3 and 4,in human endom etrium [J]. J Reprod Immunol,2003,59(1):1-16.
[14] Canciello A,Russo V,Berardinelli P,et al. Progesterone prevents epithelial-mesenchymal transition of ovine amniotic epithelial cells and enhances their immunomodulatory properties [J]. Sci Rep,2017,19:7(1):3761.