刺鼠信号蛋白通过PPARγ途径促进高糖环境下滋养层细胞的增殖、侵袭和成管能力OA
Agouti Signaling Protein Promotes Trophoblast Cell Proliferation,Invasion,and Tube Formation Through PPARγ Under High-Glucose Conditions
目的 探索刺鼠信号蛋白(agouti signaling protein,ASIP)在不同糖耐量孕妇胎盘中的表达及在高糖环境下对滋养层细胞增殖、侵袭和成管能力的影响.方法 纳入妊娠期糖尿病(gestational diabetes mellitus,GDM)孕妇和非 GDM 孕妇各20 例.应用免疫组织化学和免疫荧光染色检测胎盘组织中 ASIP 的表达和定位.采用高糖培养的人滋养层细胞系 HTR-8/SVneo 细胞模型,分别给予重组人 ASIP 蛋白与过氧化物酶体增殖物激活受体 γ(peroxisome proliferator-activated receptor γ,PPARγ)的拮抗剂 GW9662 处理,通过 MTT、Transwell 及 Matrigel 实验评估细胞增殖、侵袭与成管能力.采用蛋白质印迹和 RT-qPCR 分析相关蛋白及 mRNA 表达.结果 GDM 组胎盘重量较非 GDM 组明显增加(570 g vs 490 g,P=0.019).RT-qPCR 和蛋白质印迹结果表明 GDM 胎盘中 ASIP 的 mRNA 及蛋白表达明显升高,免疫组织化学和免疫荧光染色检测进一步证实在胎盘绒毛滋养层细胞中 ASIP 蛋白表达明显升高.在高糖环境下,HTR-8/SVneo 细胞中 ASIP 的 mRNA 及蛋白水平的表达明显增加,重组人 ASIP 蛋白(50～200 ng/mL)以剂量依赖性的方式促进 HTR-8/SVneo 细胞增殖,诱导 PPARγ、信号转导与转录激活因子 1(signal transducer and activator of transcription 1,STAT1)和 STAT3 的磷酸化.ASIP(200 ng/mL)作用 HTR-8/SVneo 细胞 24 h 后,细胞的增殖、侵袭和成管能力均显著增强,加入 PPARγ 拮抗剂 GW9662(20 μmol/L)后上述效应可被部分逆转(P均<0.01).结论 GDM/高糖环境下胎盘滋养层细胞中 ASIP 表达升高,ASIP 通过 PPARγ 依赖性机制促进高糖环境下滋养层细胞的过度增殖、侵袭及成管,从而参与 GDM 病理性胎盘重塑.
Objective To investigate the expression of agouti signaling protein(ASIP)in the placenta of pregnant women with different glucose tolerance,as well as its effect,under high-glu-cose conditions,on trophoblast cell proliferation,invasion,and tube formation.Methods Placen-tal tissues were collected from pregnant women with gestational diabetes mellitus(GDM,n=20)and without GDM(non-GDM,n=20).ASIP expression and localization were detected by immuno-histochemistry and immunofluorescence.The human trophoblast cell line HTR-8/SVneo was cultured under high-glucose conditions and treated with recombinant ASIP or the peroxisome proliferator-acti-vated receptor γ(PPARγ)antagonist GW9662.Cell proliferation,invasion,and tube formation were assessed using MTT,Transwell,and Matrigel assays,respectively.Western blotting assay and RT-qPCR were performed to analyze protein and mRNA expression.Results Placental weight was significantly higher in the GDM group than in the non-GDM group(570 g vs 490 g,P=0.019).ASIP mRNA and protein levels were significantly upregulated in GDM placental villous trophoblasts,and immunohistochemistry and immunofluorescence further confirmed significantly increased ASIP protein expression in the placental villous trophoblasts.Under high-glucose conditions,the expres-sion levels of ASIP mRNA and protein in HTR-8/SVneo cells were significantly increased.Subsequently,recombinant ASIP(50-200 ng/mL)promoted cell proliferation and in-duced the phosphorylation of PPARγ,signal transducer and activator of transcription 1(STAT1),and STAT3 in a dose-dependent manner.Furthermore,treatment of HTR-8/SVneo cells with ASIP(200 ng/mL)for 24 h significantly enhanced trophoblast proliferation,invasion,and tube forma-tion.The promoting effects of ASIP were partially reversed by co-treatment with the PPARγ antago-nist GW9662(20 μmol/L).Conclusion ASIP expression is elevated in trophoblast cells under both GDM and high-glucose conditions.ASIP promotes excessive proliferation,invasion,and tube formation of trophoblast cells via a PPARγ-dependent mechanism under high-glucose conditions,thereby contributing to the placental remodeling of GDM.
韦雪;龚浩;张倩;董书琴;韦静;卢宇
南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 内分泌科 江苏省 泰州市,225300南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 内分泌科 江苏省 泰州市,225300南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 内分泌科 江苏省 泰州市,225300南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 内分泌科 江苏省 泰州市,225300南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 妇产科 江苏省 泰州市,225300南京医科大学泰州临床医学院,南京医科大学附属泰州人民医院 内分泌科 江苏省 泰州市,225300
医药卫生
刺鼠信号蛋白滋养层细胞胎盘妊娠期糖尿病过氧化物酶体增殖物激活受体γ
agouti signaling proteintrophoblastplacentagestational diabetes mellitusperoxisome proliferator-activated receptor γ
《医学分子生物学杂志》 2026 (3)
250-257,8
江苏省"六大人才高峰"项目(No.WSN-336),江苏省卫生健康委科研项目(No.LGY2020067),南京医科大学泰州临床医学院科研项目(No.TZKY20220110) This work was supported by grants from the Six Talent Peaks Project in Jiangsu Province(No.WSN-336),Scientific Research Project of Health Commission in Jiangsu Province(No.LGY2020067),Scientific Research Project of Taizhou School of Clinical Medicine of Nanjing Medical University(No.TZKY20220110).
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