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目的探讨橘荔散结丸各有效部位对子宫肌瘤细胞孕激素受体亚型(PR-A)、表皮生长因子(EGF)、表皮生长因子受体(EGFR)表达水平的影响。方法采用胶原酶消化法对人体的子宫肌瘤细胞进行原代培养,运用现代技术提取橘荔散结丸,将不同浓度的橘荔散结丸各有效部位作用于子宫肌瘤细胞24 h后,分别采用酶联免疫法、免疫细胞化学技术及免疫荧光技术检测子宫肌瘤细胞EGF分泌变化、及EGFR和PR-A表达的改变。结果与空白对照组相比:高浓度醇提物、水提醇沉液及挥发油组EGF分泌均显著下降,差异有统计学意义(P<0.05);橘荔散结丸醇提物、水提醇沉液、水提醇沉物及挥发油组EGFR阳性表达细胞的平均光密度(MOD)均显著降低,差异有统计学意义(P<0.05),挥发油组间成浓度依赖关系(P<0.05);各橘荔散结丸有效部位组PR-A阳性表达细胞的MOD呈显著降低,差异有统计学意义(P<0.05)。与阳性对照米非司酮(RU486)组相比:橘荔散结丸的醇提物、水提醇沉液、水提醇沉物及挥发油的高浓度组的EGFR表达显著降低,差异有统计学意义(P<0.05)。结论本研究表明橘荔散结丸各有效部位可降低子宫肌瘤细胞EGF的分泌或调控子宫肌瘤细胞PR-A、EGFR的表达,各有效部位协同作用可能是橘荔散结丸治疗子宫肌瘤的主要机理之一,并且橘荔散结丸各有效部位对EGFR的降调作用优于RU486,为药物筛选、精简处方奠定基础。
Objective To investigate the effect of various effective sites of the tangerine Lai Sanjie Pill on the expression of progesterone receptor subtypes (PR-A), epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in uterine fibroids. Methods Human uterine leiomyoma cells were primarily cultured by collagenase digestion method. Tumor samples of Tumor were extracted by using modern technology. Twenty-four hours after different concentrations of Triclosan, Enzyme-linked immunosorbent assay, immunocytochemistry and immunofluorescence were used to detect the change of EGF secretion and the expression of EGFR and PR-A in uterine fibroids. Results Compared with the blank control group, EGF secretion was significantly decreased in high alcohol extract, aqueous alcohol extract and volatile oil, the difference was statistically significant (P <0.05) The average optical density (MOD) of EGFR-positive cells in alcohol-precipitated liquid, alcohol-precipitated water extract and volatile oil group were significantly decreased (P <0.05) (P <0.05). The MOD of PR-A positive cells in the effective sites of each group were significantly decreased (P <0.05). Compared with the RU486 positive control group, EGFR expression was significantly decreased in the alcohol extract, water-extract and alcohol-sediment solution, water-extractable alcohol precipitates, and high-concentration group of volatile oil, with statistically significant differences Significance (P <0.05). Conclusions This study shows that the effective fractions of taijiujiejie pill can reduce the secretion of EGF in uterine leiomyoma cells or regulate the expression of PR-A and EGFR in uterine leiomyoma cells. The synergistic effect of each effective site may be that the treatment of uterine muscle Tumor one of the main mechanism, and the role of each part of the orange litchi Sanjie pill EGFR is better than RU486, for drug screening, to lay a foundation for streamlining prescription.