资冲颗粒含药血清对ανβ3、HLA-G、HOXA10、Fas/FasL、IDO在人早孕绒毛膜组织中表达的影响

Effects of rat medicated serum containing Zichong Granule on expressions of ανβ3, HLA-G, HOXA10, Fas/FasL and IDO in the human first trimester chorionic tissue culture in vitro

目的:探讨资冲颗粒对体外人早孕绒毛膜组织ανβ3、HLA-G、HOXA10、Fas/Fas L和IDO基因表达的影响。方法:对体外培养的人早孕绒毛膜组织加入2%、5%、10%的鼠资冲颗粒含药血清培养24-72h。同时,在组织培养中加入孕酮作为阳性对照组而仅有基础培养基为空白组。采用Q-PCR技术检测ανβ3、HLA-G、HOXA10、Fas/Fas L和IDO m RNA的表达。采用特异ELISA试剂盒定量检测ανβ3和HLA-G的蛋白表达。结果:10%含药血清处理的绒毛膜组织的ανβ3、HLA-G、HOXA10、Fas/Fas L、IDO基因表达均显著高于阳性对照组及空白组(P<0.05)。10%含药血清处理的绒毛膜组织的ανβ3和HLA-G蛋白表达显著高于阳性对照组及空白组(P<0.05)。结论:资冲颗粒具有增加ανβ3、HLA-G、HOXA10、Fas/Fas L和IDO基因表达的作用,可能是该中药复方治疗不孕症分子生物学机制之一。

Objective： To explore the effects of rat medicated serum containing Zichong Granule（ZCG） on the expressions of ανβ3, HLA-G, HOXA10, Fas/Fas L and IDO in the first trimester chrionic tissue culture in vitro. Methods： The first trimester chorionic tissues were cultured and treated with different concentrations of rat medicated serums containing ZCG in varying couture time（2%, 5% or 10%; 24-72h）. At the same time, cultures treated with progesterone were used as positive control while cultures without any treatment were used as blank control. Transcriptions of HLA-G, HOXA10, Fas/Fas L and IDO were determined by using quantitative real-time RT-PCR（q RT-PCR） while translations of HLA-G and ανβ3 were detected by using specific HLA-G or ανβ3 enzyme-linked immunosorbent assays（ELISA）. Results： Compared with the positive and the blank groups, m RNA levels of ανβ3, HLA-G, HOXA10, Fas/Fasl and IDO in 10% rat medicated serum were significantly increased（P〈0.05）. Further, ανβ3 and HLA-G protein levels in 10% medicated serum were also increased notably（P〈0.05）. Conclusion： ZCG had an up-regulated effect on gene expressions of ανβ3, HLA-G, HOXA10, Fas and IDO in first trimester chorionic tissue in vitro. It might be one of molecular mechanisms for the formula in treatment of infertility.