【摘要】 目的:观察不同浓度的安胎协定方含药血清对离体人早孕绒毛滋养细胞HTR-8细胞增殖活力及ERK1/2信号通路的影响,探究其维持妊娠机制。方法:雌性SD大鼠灌胃制备空白血清和各组含药血清。将HTR-8细胞分为5组:空白对照组,地屈孕酮组,中药高、中、低剂量组(5%、10%、20%含药血清)。MTT法检测24、48、72h的吸光度,表示细胞增殖活力,免疫组化法检测48h细胞EGF、EGRR及ERK1/2表达含量。结果:与空白对照组比较,除中药低剂量组外,其余各组3个时间段的吸光度及48h细胞EGF、EGFR、ERK1/2含量均显著升高(P<0.05);与地屈孕酮组比较,中药低剂量组3个时间段的吸光度及48h细胞EGF、EGFR、ERK1/2含量显著降低(P<0.05);与中药低剂量组比较,中药中、高剂量组3个时间段的吸光度及48h细胞EGF、EGFR、ERK1/2含量显著升高(P<0.05)。结论:安胎协定方可改善绒毛滋养细胞增殖活力,避免增殖过度以维持妊娠,可能是通过适度上调细胞内EGF及EGFR的含量表达,促进ERK1/2磷酸化而实现的。更多还原

【Abstract】 Objective: To investigate the effect of serum containing different concentrations of Antai Xieding Formula on the proliferation of the human trophoblast cell HTR-8 in vitro, and ERK1/2 signaling pathway, and then to explore the mechanism of pregnancy maintenance. Methods: Female SD rats were perfused to prepare blank serum and drug containing serum each group. HTR-8 cells were divided into 5 groups: blank control group, the dydrogesterone group, high dose, middle and low dose group(5%, 10%, 20% drug containing serum). The absorbance of 24, 48 and 72 h was detected by MTT, and the proliferation activity of cells was measured. The expression level of EGF, EGRR and ERK1/2 in 48 h cells was detected by immunohistochemistry. HTR-8 cells can be divided into 5 groups: the blank control group, the dydrogesterone control group, Antaixieding formula high, medium and low dose group; low dose group(5% medicated serum), middle dose group(10% medicated serum), high dose group(20% medicated serum). MTT method was used to detect the absorbance of 24 h, 48 h and 72 h in three time periods, which indicated the cell proliferation activity. Immunohistochemistry method was used to detect the expression of EGF, EGRR and ERK1/2. Results: Compared with the blank control group, except the low dose group, others are statistically significant among the three periods(P<0.05); In the expression of EGF, EGFR and ERK1/2, the medium and high dose group are significantly increased(P<0.05); Compared with the dydrogesterone group, the low dose group has statistically significant among the three periods, the expression of EGF, EGFR and ERK1/2 are significantly decreased(P<0.05). Compared with low dose group, the absorbance in three periods and 48 h EGF, EGFR, ERK1/2 expression in high and middle dose group have significantly increased(P<0.05). Conclusion: Antaixieding formula can effectively maintain pregnancy by improving the cell proliferation activity. The mechanism may be achieved by appropriately upregulating the expression of EGF and EGFR in the cell and promoting the phosphorylation of ERK1/2.更多还原