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目的探讨加味芍药甘草汤及其含药血清治疗子宫腺肌病的深层作用机制。方法选取6例行全宫切除术的子宫腺肌病患者子宫病灶组织进行体外原代细胞培养,所培养病灶细胞采用免疫细胞化学法鉴定。将实验分为加味芍药甘草汤高浓度(20 g/L)及低浓度(10 g/L)组、加味芍药甘草汤含药血清组(简称含药血清组)、空白血清组、米非司酮组及空白对照组6组,各组药物分别干预病灶细胞24 h和48 h后采用酶联免疫吸附法(ELISA)检测各组细胞上清雌激素(E2)的表达,采用免疫细胞化学法检测各组细胞雌激素受体(ER)及芳香化酶P450(P450arom)的表达。结果经免疫细胞化学法鉴定:共收集及培养成功的6例细胞均为子宫腺肌病病灶细胞。与空白对照组及空白血清组相比,含药血清组可明显降低细胞E2水平(P<0.05);加味芍药甘草汤高低浓度组、含药血清组、米非司酮组24 h均可明显降低细胞ER水平(P<0.05),其中米非司酮组总体降ER效果劣于加味芍药甘草汤高低浓度组及含药血清组(P<0.05),加味芍药甘草汤高浓度组24 h降ER作用优于48 h(P<0.05)。与空白血清及空白对照组相比,加味芍药甘草汤高低浓度组、含药血清组、米非司酮组24 h细胞P450arom水平明显降低(P<0.05),且加味芍药甘草汤高浓度、低浓度24 h及含药血清24 h作用较米非司酮好(P<0.05),加味芍药甘草汤低浓度组24 h作用优于48 h(P<0.05)。总之,加味芍药甘草汤降ER及P450arom作用较米非司酮好,效果呈时效性,24 h优于48 h,无浓度依赖性。结论通过中药复方水提液及动物含药血清两种方式证实加味芍药甘草汤能降低子宫腺肌病病灶细胞ER、P450arom水平,阻断E2、P450arom正反馈环、弱化E2效应,可能是其治疗子宫腺肌病的机制之一。
Objective To investigate the underlying mechanism of Jiawei Shaoyao Gancao Decoction and its medicated serum in the treatment of adenomyosis. Methods Six cases of uterine lesions of patients with adenomyosis underwent hysterectomy were selected for primary culture in vitro. Immunocytochemistry was used to identify the cells in culture. The experiment was divided into Jiawei Shaoyao Gancao Decoction high concentration (20 g / L) and low concentration (10 g / L) group, Jiawei Shaoyao Gancao Decoction containing serum group (referred to as containing serum group), blank serum group, Ketone group and blank control group. Each group of drugs intervened the lesion cells for 24 h and 48 h, respectively. The expression of estrogen (E2) in the supernatant of each group was detected by enzyme linked immunosorbent assay (ELISA). Immunocytochemistry The estrogen receptor (ER) and aromatase P450 (P450arom) expression in each group were detected. The results identified by immunocytochemistry: a total of 6 cases were successfully collected and cultured cells of adenomyosis lesions. Compared with the blank control group and the blank serum group, the drug-containing serum group could obviously reduce the E2 level (P <0.05). The Jiawei Shaoyao Gancao Decoction high concentration group, the drug-containing serum group and the mifepristone group could all obviously (P <0.05). The overall effect of ER in mifepristone group was lower than that in Jiawei Shaoyao Gancao Decoction group and the drug-containing serum group (P <0.05), while Jiawei Shaoyao Gancao Decoction high concentration group ER was better than 48 h (P <0.05). Compared with the blank serum and the blank control group, the P450arom level in the Jiawei Shaoyao Gancao Decoction high and low concentration group, the drug-containing serum group and the mifepristone group was significantly decreased (P <0.05), and the Jiawei Shaoyao Gancao Decoction high concentration and low Concentration of 24 h and serum containing 24 h than mifepristone good (P <0.05), Jiawei Shaoyao Gancao Tang low concentration group 24 h better than 48 h (P <0.05). In conclusion, Jiawei Shaoyao Gancao Decoction decreased the effect of ER and P450arom compared with mifepristone, with a time-effect effect, which was superior to 48 h in 24 h, without concentration-dependent. CONCLUSION: Jiawei Shaoyao Gancao Decoction can reduce ER and P450arom levels in the adenomyosis cells and block the E2 and P450arom positive feedback loops and weaken the E2 effect, which may be due to the treatment of the traditional Chinese medicine compound aqueous extract and animal serum-containing serum One of the mechanisms of adenomyosis.