小檗碱对胰岛素抵抗卵巢甾体激素体外合成和代谢的调控作用

作者	单位
王秀秀	黑龙江中医药大学附属第一医院妇产科
李威	黑龙江中医药大学附属第一医院妇产科
刘逸超	黑龙江中医药大学附属第一医院妇产科
侯丽辉	黑龙江中医药大学附属第一医院妇产科
吴效科	黑龙江中医药大学附属第一医院妇产科

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中文摘要:

目的研究体外培养的胰岛素抵抗卵巢模型的功能和代谢变化,观察具有胰岛素增敏作用中药单体小檗碱对胰岛素抵抗的改善情况。方法体外培养小鼠卵巢,地塞米松(dexamethasone,Dex)诱导卵巢胰岛素抵抗模型,通过检测培养液葡萄糖含量、激素水平及逆转录-聚合酶链反应(RT-PCR)评估胰岛素抵抗模型卵巢功能的变化情况,检测胰岛素通路及激素合成通路关键分子的表达情况以及小檗碱对胰岛素抵抗的改善情况。结果(1)Dex诱导卵巢胰岛素抵抗的效果存在时间剂量依赖性,经浓度为300nmol/LDex处理48h后,卵巢对葡萄糖的摄取量[(2.48±0.29)mg/g]较未用Dex培养卵巢48h组[(9.05±0.75)mg/g]下降(P<0.05);在胰岛素刺激下,Dex处理的卵巢对葡萄糖的摄取量[(1.94±0.19)mg/g]下降,与未用Dex处理72h组[(9.59±1.74)mg/g]比较,差异有统计学意义(P<0.01),卵巢胰岛素抵抗模型理想。应用中药小檗碱后,卵巢葡萄糖摄取量[(13.95±3.30)mg/g]较模型组[(1.89±0.33)mg/g]明显升高(P<0.05)。(2)模型组培养液中睾酮(T)和雄烯二酮(A2)水平升高,孕酮(P)和17-羟孕酮(17-OHP)水平明显下降,小檗碱能显著进一步降低T和A2浓度,并使得培养液17-OHP水平升高,但对P水平影响不大。(3)RT-PCR结果显示:Dex诱导后,卵巢17-羟化酶(CYP17)、极微染色体维持蛋白(MCM-2)的mRNA表达水平升高,细胞外信号调节激酶-1(ERK-1)、蛋白激酶B(AKT-2)、糖原合酶激酶-3(GSK-3β)的mRNA表达水平下降,应用小檗碱后,上述分子指标明显逆转。结论(1)Dex能够诱导卵巢器官胰岛素抵抗,卵巢产生胰岛素抵抗对雄激素合成可能存在促进作用。(2)中药小檗碱能降低卵巢的胰岛素抵抗程度和雄激素合成,提示对胰岛素有增敏作用的中药,对多囊卵巢综合征有良好的治疗作用。

英文摘要:

Objective To investigate the functional and metabolic alterations in cultured insulin resistant ovary model in vitro, and to observe the effect of berberine (Ber, a Chinese medical monomer) in improving insulin resistance (IR). Methods Ovary of mouse was cultured in vitro and treated by dexamethasone (Dex) to induce IR for establishing IR model ovary. The functional alteration in model ovary was assessed through detecting glucose and hormone levels in medium using RT-PCR, meanwhile, the expression of key molecules in insulin signal and steroid synthetic pathway were detected, and condition of IR improved by berberine was evaluated also. Results (1) The model ovary was made by Dex in dose- and acting time-dependent manner. After being treated by 300 nmol/L Dex for 48 h, the glucose uptake of ovary reduced from 9.05±0.75 mg/g to 2.48±0.29 mg/g (P<0.05); it further decreased (from 9.59±1.74 mg/g to 1.94±0.19 mg/g, P<0.01) under the stimulation of insulin, which proved that the IR model ovary was made successfully. Berberine significantly increased the glucose uptake of model ovaries (1.89±0.33 mg/g to 13.95±3.30 mg/g,P<0.05). (2) As compared with control group, levels of testosterone (T) and androstenedione (A2) were higher, and levels of progesterone (P) and 17-hydroxyprogesterone (17-OHP) were lower significantly in the model. Berberine reversed the alternations of T, A2 and 17-OHP levels, but did not influence the level of P. (3) RT-PCR showed that the mRNA expressions of cytochrome 17-hydroxylase (CYP17) and mini-chromosome maintenance protein-2 (MCM-2) elevated, but extracellular regulated protein-1 (ERK-1), protein kinase B (AKT-2) and glycogen synthase kinase-3 (GSK-3β) lowered in the medium after Dex inducing. Berberine treatment restored these molecular index obviously. Conclusions (1) Dex could induce IR in mouse ovary, which might enhance the androgenic synthesis. (2) Berberine could alleviate the degree of IR and the androgen synthesis, indicating that the Chinese sensitizing agents has favorable therapeutic effect for the treatment of polycystic ovaries.

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