Objective To investigate the deep mechanism of Jiawei Shaoyao Gancao Tang (Modified Peony and Licorice Root Decoction) and its medicated serum in treatment of adenomyosis.Methods The cells in ovarian focus tissue were given in vitro primary culture in 6 patients with adenomyosis undergone hysterectomy,and the focus cells were identified by using immunoeytochemistic method.There were 6 groups in this study including high-dose Jiawei Shaoyao Gancao Tang group (high-dose JSGT group,20 mg/mL),low-dose Jiawei Shaoyao Gancao Tang group (low-dose JSGT group,10 mg/mL),Jiawei Shaoyao Gancao Tang medicated serum group (medicated serum group),blank serum group,mifepristone group and blank control group.The focus cells were intervened for 24 h and 48 h respectively in all groups,and then the expression of estrogen (E2) was detected by using enzyme-linked immunosorbent assay (ELISA).The expressions of estrogen receptor (ER) and aromatase P450 (P450arom) were detected by using immunocytochemistic method in all groups.Results The identification results of immunocytochemistic method showed that collected from 6 patients and successfully cultured cells were all focus cells of adenomyosis.The level of E2 decreased significantly in medicated serum group compared with blank control group and blank serum group (P ＜ 0.05).The level of ER decreased significantly in high-dose JSGT group,low-dose JSGT group,medicated serum group and mifepristone group after 24 h (P ＜ 0.05).The total efficacy of decreasing ER was inferior in mifepristone group to that in high-dose JSGT group,low-dose JSGT group and medicated serum group (P ＜ 0.05),and efficacy of decreasing ER was superior after 24 h to that after 48 h in high-dose JSGT group (P ＜ 0.05).The level of P450arom decreased significantly in high-dose JSGT group,low-dose JSGT group,medicated serum group and mifepristone group after 24 h compared with blank serum group and blank control group (P ＜ 0.05),and the efficacy decreasing P450arom was better in high-dose JSGT group,low-dose JSGT group and medicated serum group than that in mifepristone group after 24 h (P ＜ 0.05).The efficacy decreasing P450arom was superior after 24 h to that after 48 h in low-dose JSGT group (P ＜ 0.05).Jiawei Shaoyao Gancao Tang's efficacies of decreasing ER and P450arom were better than mifepristone and these efficacies showed timeliness:the efficacies were better after 24 h than that after 48 h.There was no dose dependence.Conclusion Jiawei Shaoyao Gancao Tang can reduce the levels of ER and P450arom,cut off the positive feedback loop of E2 and P450arom and weaken E2 effect,which may be its mechanism in treatment of adenomyosis.%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.
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