中国中西医结合杂志
中國中西醫結閤雜誌
중국중서의결합잡지
CHINESE JOURNAL OF INTEGRATED TRADITIONAL AND WESTERN MEDICINE
2010年
2期
161-166
,共6页
王秀秀%李威%刘逸超%侯丽辉%吴效科
王秀秀%李威%劉逸超%侯麗輝%吳效科
왕수수%리위%류일초%후려휘%오효과
胰岛素抵抗%多囊卵巢综合征%卵巢%小檗碱
胰島素牴抗%多囊卵巢綜閤徵%卵巢%小檗堿
이도소저항%다낭란소종합정%란소%소벽감
insulin resistance%polycystic ovarian syndrome%ovary%berberine
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300 nmol/L Dex处理48 h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48 h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72 h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(A_2)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A_2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.
目的 研究體外培養的胰島素牴抗卵巢模型的功能和代謝變化,觀察具有胰島素增敏作用中藥單體小檗堿對胰島素牴抗的改善情況.方法 體外培養小鼠卵巢,地塞米鬆(dexamethasone,Dex)誘導卵巢胰島素牴抗模型,通過檢測培養液葡萄糖含量、激素水平及逆轉錄-聚閤酶鏈反應(RT-PCR)評估胰島素牴抗模型卵巢功能的變化情況,檢測胰島素通路及激素閤成通路關鍵分子的錶達情況以及小檗堿對胰島素牴抗的改善情況.結果 (1) Dex誘導卵巢胰島素牴抗的效果存在時間劑量依賴性,經濃度為300 nmol/L Dex處理48 h後,卵巢對葡萄糖的攝取量[(2.48±0.29)mg/g]較未用Dex培養卵巢48 h組[(9.05±0.75)mg/g]下降(P<0.05);在胰島素刺激下,Dex處理的卵巢對葡萄糖的攝取量[(1.94±0.19)mg/g]下降,與未用Dex處理72 h組[(9.59±1.74)mg/g]比較,差異有統計學意義(P<0.01),卵巢胰島素牴抗模型理想.應用中藥小檗堿後,卵巢葡萄糖攝取量[(13.95±3.30)mg/g]較模型組[(1.89±0.33)mg/g]明顯升高(P<0.05).(2)模型組培養液中睪酮(T)和雄烯二酮(A_2)水平升高,孕酮(P)和17-羥孕酮(17-OHP)水平明顯下降,小檗堿能顯著進一步降低T和A_2濃度,併使得培養液17-OHP水平升高,但對P水平影響不大.(3)RT-PCR結果顯示:Dex誘導後,卵巢17-羥化酶(CYP17)、極微染色體維持蛋白(MCM-2)的mRNA錶達水平升高,細胞外信號調節激酶-1(ERK-1)、蛋白激酶B(AKT-2)、糖原閤酶激酶-3(GSK-3β)的mRNA錶達水平下降,應用小檗堿後,上述分子指標明顯逆轉.結論 (1)Dex能夠誘導卵巢器官胰島素牴抗,卵巢產生胰島素牴抗對雄激素閤成可能存在促進作用.(2)中藥小檗堿能降低卵巢的胰島素牴抗程度和雄激素閤成,提示對胰島素有增敏作用的中藥,對多囊卵巢綜閤徵有良好的治療作用.
목적 연구체외배양적이도소저항란소모형적공능화대사변화,관찰구유이도소증민작용중약단체소벽감대이도소저항적개선정황.방법 체외배양소서란소,지새미송(dexamethasone,Dex)유도란소이도소저항모형,통과검측배양액포도당함량、격소수평급역전록-취합매련반응(RT-PCR)평고이도소저항모형란소공능적변화정황,검측이도소통로급격소합성통로관건분자적표체정황이급소벽감대이도소저항적개선정황.결과 (1) Dex유도란소이도소저항적효과존재시간제량의뢰성,경농도위300 nmol/L Dex처리48 h후,란소대포도당적섭취량[(2.48±0.29)mg/g]교미용Dex배양란소48 h조[(9.05±0.75)mg/g]하강(P<0.05);재이도소자격하,Dex처리적란소대포도당적섭취량[(1.94±0.19)mg/g]하강,여미용Dex처리72 h조[(9.59±1.74)mg/g]비교,차이유통계학의의(P<0.01),란소이도소저항모형이상.응용중약소벽감후,란소포도당섭취량[(13.95±3.30)mg/g]교모형조[(1.89±0.33)mg/g]명현승고(P<0.05).(2)모형조배양액중고동(T)화웅희이동(A_2)수평승고,잉동(P)화17-간잉동(17-OHP)수평명현하강,소벽감능현저진일보강저T화A_2농도,병사득배양액17-OHP수평승고,단대P수평영향불대.(3)RT-PCR결과현시:Dex유도후,란소17-간화매(CYP17)、겁미염색체유지단백(MCM-2)적mRNA표체수평승고,세포외신호조절격매-1(ERK-1)、단백격매B(AKT-2)、당원합매격매-3(GSK-3β)적mRNA표체수평하강,응용소벽감후,상술분자지표명현역전.결론 (1)Dex능구유도란소기관이도소저항,란소산생이도소저항대웅격소합성가능존재촉진작용.(2)중약소벽감능강저란소적이도소저항정도화웅격소합성,제시대이도소유증민작용적중약,대다낭란소종합정유량호적치료작용.
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 (A_2) were higher,and levels of progesterone (P) and 17-hydroxyprogesterone (17-OHP) were lower significantly in the model. Berberine reversed the alternations of T,A_2 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.
