化瘀止痛方及其拆方对人子宫腺肌病细胞OTR mRNA、ERα mRNA的影响

Influence of Huayu Zhitong Prescription and Its Separate Components on OTR mRNA and ERα mRNA of Human Adenomyosis Cells

目的观察化瘀止痛方及其拆方对人子宫腺肌病细胞缩宫素受体(OTR)mRNA、雌激素受体α(ERα)mRNA表达的影响。方法将化瘀止痛方及其拆方分为化瘀止痛方高、低剂量组,缓急止痛方高、低剂量组,活血行气方高、低剂量组,另设空白对照组、基质组及米非司酮(RU486)组,每组5例,采用胶原酶消化法培养人子宫腺肌病细胞,采用荧光RT-PCR检测药物干预前后子宫腺肌病病灶细胞OTR mRNA、ERαmR-NA表达的变化。结果与空白对照组比较,各给药组子宫腺肌病细胞的OTR mRNA表达均有不同程度下降,化瘀止痛方高、低剂量组及活血行气高剂量组的OTR mRNA表达显著下降,差异均有统计学意义(P<0.05);与RU486组比较,化瘀止痛方高、低剂量组OTR mRNA表达均有不同程度下降,差异均有统计学意义(P<0.05);与缓急止痛低剂量组比较,化瘀止痛方高、低剂量组OTR mRNA表达显著下降,差异有统计学意义(P<0.05)。与空白对照组比较,各给药组细胞的ERαmRNA表达均有不同程度的下降,化瘀止痛方高、低剂量组及缓急止痛高、低剂量组的ERαmRNA表达显著下降,差异有统计学意义(P<0.05)。与RU486对照组比较,化瘀止痛方高、低剂量组、缓急止痛高剂量组子宫腺肌病细胞ERαmRNA表达显著下降,差异有统计学意义(P<0.05)。化瘀止痛方高剂量组与活血行气高剂量组比较,子宫腺肌病细胞ERαmRNA表达显著下降,差异有统计学意义(P<0.05)。结论化瘀止痛方可能通过降低子宫腺肌病病灶细胞OTR mRNA、ERαmRNA表达,减缓子宫平滑肌的收缩强度,从而缓解患者的痛经症状,在降低子宫腺肌病病灶细胞OTR mRNA表达方面活血行气药贡献值较大,在子宫腺肌病病灶细胞ERαmRNA表达变化中缓急止痛药的贡献值较大。

Objective To explore the influence of Huayu Zhitong Prescription （HZP） and its separate components on OTR mRNA and ERa mRNA of the human adenomyosis cells. Methods Adenomyosis cells were digested with collagenase in primary culture. Nine groups were set up for the experiment, and they were high- and low-dose HZP groups, high- and low-dose groups of spasm-relieving and pain-stopping herbs, high- and low-dose groups of blood-activating and Qi-promoting herbs, blank control group, basic medium group, and positive control group of mifepristone （RU486） . Before and after drug intervention, the expression levels of OTR mRNA and ERa mRNA in the human adenomyosis cells were detected by fluorescence RT-PCR. Results Compared with the blank control group, the expression of OTR mRNA was decreased in the medication groups to various degrees, and a significant decrease was shown in HZP groups and high-dose blood-activating and Qi-promoting group（P 〈 0.05）. There was significant difference between HZP groups and RU486 group in the expression of OTR mRNA（P 〈 0.05）. Compared with low-dose spasm-relieving and pain-stopping group, HZP groups had an obvious effect on decreasing OTR mRNA expression（P 〈 0.05）. As for the expression of EroL mRNA, HZP groups and high-dose spasm-relieving and pain-stopping group had lower level than the blank control group and RU486 group（P 〈 0.05）, high-dose HZP group had lower level than high-dose blood-activating and Qi-promoting group（P 〈 0.05）. Conclusion HZP down-regulates the expression of OTR mRNA and ER mRNA in adenomyosis cells, which reduces the intensity of uterine smooth muscle contraction, and effectively alleviate the symptoms of patients with dysmenorrhea. Blood-activating and Qi-promoting herbs of HZP may be the main effective components in decreasing OTR mRNA, and spasm-relieving and pain-stopping herbs bene- fit to the change of ERα mRNA of the adenomyosis cells.