过氧化物酶体增殖物激活受体γ及其配体对早孕期绒毛组织及细胞滋养细胞浸润能力的影响

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目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)及其配体对早孕期绒毛组织及细胞滋养细胞浸润能力的影响。方法采用免疫组化方法、免疫荧光细胞化学染色法、蛋白印迹法和RT-PCR 技术检测20例孕6~8周(早孕早期组)及20例孕11~12周(早孕晚期组)绒毛组织及细胞滋养细胞中的 PPARγ蛋白及其 mRNA 的表达;并检测不同浓度 PPARγ激动配体——15-脱氧-前列腺素 J_2(15-d-PGJ_2)和曲格列酮,以及不同浓度拮抗配体——双酚丙烷二环氧甘油醚(BADGE)对原代无血清培养的细胞滋养细胞浸润能力的影响。结果 (1)PPARγ蛋白在早孕早期组和早孕晚期组绒毛组织中均有表达,主要定位在细胞滋养细胞核中,合体滋养细胞及绒毛问质细胞中无表达。(2)早孕早期组绒毛组织和培养的细胞滋养细胞中,PPARγ蛋白表达水平分别为1.35±0.08、1.13±0.11,PPARγ mRNA 表达水平分别为36.0±5.1、13.4±3.1;早孕晚期组绒毛组织和培养的细胞滋养细胞中,PPARγ蛋白表达水平分别为1.17±0.03、0.86±0.05,PPARγ mRNA 表达水平分别为23.3±5.5、6.1±1.3,早孕晚期组 PPARγ蛋白及其 mRNA 表达水平明显低于早孕早期组,两组分别比较,差异均有统计学意义(P<0.05)。(3)PPARγ激动配体15-d-PGJ_2和曲格列酮均有抑制细胞滋养细胞的浸润的作用。15-d-PGJ_2浓度为1、10μmoL/L,曲格列酮浓度为10μmoL/L 时,早孕早期组细胞滋养细胞浸润指数分别为0.57±0.03、0.43±0.02、0.50±0.06,早孕晚期组分别为0.69±0.02、0.59±0.03、0.66±0.05,两组分别比较,差异均有统计学意义(P<0.05)。(4)PPARγ拮抗配体 BADGE 浓度为20、50μmol/L 时,早孕早期组细胞滋养细胞浸润指数分别为1.23±0.07和1.58±0.04;早孕晚期组分别为1.05±0.03和1.38±0.08,两组分别比较,差异均有统计学意义(P<0.05)。结论 PPARγ在调节滋养细胞浸润过程中起重要作用;在早孕期胎盘绒毛组织,PPARγ激动配体可抑制滋养细胞浸润;PPARγ拮抗配体可促进滋养细胞浸润,且能部分逆转激动配体的作用。 Objective To investigate the effect of peroxisome proliferator - activated receptor γ (PPARγ) and its ligand on the infiltration capacity of chorionic villi and cytotrophoblasts in early pregnancy. Methods Immunohistochemistry, immunofluorescence cytochemistry, Western blotting and RT-PCR were used to detect the expression of chorionic villi in 20 pregnant women aged 6-8 weeks (early pregnancy) and 20 cases of pregnant 11-12 weeks (early pregnancy) And PPARγprotein and its mRNA expression in cytotrophoblasts were detected. Different concentrations of 15-deoxy-prostaglandin J_2 (15-d-PGJ_2) and troglitazone Effect of Bisphenol Propane Diglycidyl Ether (BADGE) on the Infiltration of Cytotrophoblasts in Primary Serum - free Culture. Results (1) The expression of PPARγ protein in early pregnancy early pregnancy group and early late pregnancy villus tissue were mainly located in the nucleus of trophoblastic cells, syncytiotrophoblast cells and villous seminiferous cells were not expressed. (2) The expression of PPARγprotein in early chorionic villi and cultured cells were 1.35 ± 0.08 and 1.13 ± 0.11 respectively, and the mRNA expression levels of PPARγ were 36.0 ± 5.1 and 13.4 ± 3.1 respectively. The level of PPARγ protein in cultured cytotrophoblasts were 1.17 ± 0.03,0.86 ± 0.05 and 23.3 ± 5.5,6.1 ± 1.3, respectively. The expression levels of PPARγ protein and mRNA in late pregnancy were significantly lower than those in early pregnancy Group, the two groups were compared, the differences were statistically significant (P <0.05). (3) PPARγ agonistic ligands 15-d-PGJ_2 and troglitazone all inhibit the infiltration of cytotrophoblasts. When the concentration of 15-d-PGJ_2 was 1, 10μmoL / L and the concentration of troglitazone was 10μmoL / L, the infiltration index of cytotrophoblast in early pregnancy were 0.57 ± 0.03, 0.43 ± 0.02 and 0.50 ± 0.06, respectively 0.69 ± 0.02,0.59 ± 0.03,0.66 ± 0.05. There was significant difference between the two groups (P <0.05). (4) When the BADGE concentration of PPARγ antagonist ligand was 20 and 50μmol / L, the infiltration index of cytotrophoblast in early pregnancy was 1.23 ± 0.07 and 1.58 ± 0.04, respectively. The first trimester pregnancy group was 1.05 ± 0.03 and 1.38 ± 0.08, Respectively, the differences were statistically significant (P <0.05). Conclusion PPARγ plays an important role in the regulation of trophoblast invasion. In the placenta of early pregnancy, PPARγ agonist ligand can inhibit trophoblast invasion. PPARγ antagonist ligand can promote trophoblast invasion and partially reverse the effect of agonistic ligand.
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