补肾安胎方对人绒毛膜外滋养层细胞HTR-8分泌功能及HLA-G基因表达的调控作用

Regulatory Effect of Bushen Antai Recipe on the Secretion Function and HLA-G Expression of Human Extravillous Cytotrophoblasts HTR-8

    摘要
  • 摘要: 目的 观察补肾安胎方对人绒毛膜外滋养层细胞HTR-8分泌功能及人类白细胞抗原G（HLA-G）基因表达的调控，探讨其免疫保胎机制。方法 雌性SD大鼠灌胃制备空白血清和补肾安胎方含药血清。将人绒毛膜外滋养层细胞HTR-8细胞分为4组：空白对照组、低剂量组（5%中药血清组）、中剂量组（10%中药血清组）、高剂量组（20%中药血清组），培养24 h后，ELISA法测定培养上清液中雌二醇（E2）、孕激素(P）、人绒毛膜促性腺激素(HCG)及辅助性T细胞（Th）调控因子白介素-4(IL-4) 和干扰素 -γ（IFN-γ）水平，RT-PCR法测定HLA-G mRNA表达。结果 与空白对照组相比，低剂量组在促进E2、P分泌方面无统计学意义（P>0.05），中、高剂量组可有效提高细胞上清E2、P含量，且高剂量组高于中剂量组（P<0.05）。3组含药血清均能明显提高滋养细胞HCG的分泌水平（P<0.05），3组间差异无显著性意义（P>0.05）。低剂量组即可促进Th2型细胞因子IL-4分泌，但差异无统计学意义（P>0.05），中、高剂量组可有效促进Th2型细胞因子IL-4含量，组间差异无统计学意义（P>0.05）。3组含药血清均能显著抑制Th1型细胞因子IFN-γ（P<0.05），中、高剂量组均可增加HLA-G mRMA的表达，有显著差异（P<0.05），且高剂量组高于中剂量组（P<0.05）。结论 补肾安胎方含药血清可以改善滋养细胞分泌功能，使母胎免疫向Th2型转化，并可能部分通过上调HLA-G基因表达实现。

     

    Abstract: OBJECTIVE To study the effects of Bushen Antai Recipe (BAR) on the secretion function and HLA-G expression of human extravillous cytotrophoblasts HTR-8. METHODS SD female rats were treated with BAR or normal saline to prepare for BAR-containing serum or control serum. After centrifugation of the serum， HTR-8 were randomly divided into 4 groups： control group， low-dose BAR (BAR-L) group， medium dose BAR (BAR-M) group and high dose BAR (BAR-H) group. The BAR-L， BAR-M and BAR-H group were treated with 5%， 10% or 20% BAR-containing serum， respectively. The supematants were harvested at 24 h of the culture， and then the levels of hormone， including E2 (estradiol)， progesterone (P) and human chorionic gonadotropin (HCG)， and regulatory factors of Helper T cells (Th)， including interleukin -4 (IL-4) and interferon gamma (IFN-γ)， were measured by ELISA in culture supernatant. The mRNA of HLA-G was analyzed using quantitative RT-PCR. RESULTS The BAR-L group could promote the secretion of E2 and P with no statistically significant difference， the BAR-M group and BAR-H group could effectively improve the secretion of E2 and P， and the BAR-H group is higher than the BAR-M group（P<0.05）.The three groups could improve the HCG secretion effectively（P<0.05），while there were no significant difference among them（P>0.05）. The BAR-L group could promote the secretion of IL-4 with no statistically significant difference（P>0.05）.The BAR-M group and BAR-H group could effectively improve the IL-4 secretion， while there were no significant difference between them（P>0.05）. And the three groups could down-regulate the IFN-γ secretion effectively（P<0.05）. The BAR-M group and high dose group could improve the HLA-G expression effectively（P<0.05)， and the BAR-H group is higher than the BAR-M group（P<0.05）. CONCLUSION The BAR could elevate the secretion in human extravillous cytotrophoblasts and result in the bias response for Th2 type cytokine at the maternal-fetal interface， which might be partially achieved through up-regulating HLA-G expression.

     

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