隐丹参酮对小鼠卵巢胰岛素抵抗调控的机制研究

Effects of cryptotanshinone on ovaries insulin resistance

目的 通过地塞米松体外诱导卵巢器宫胰岛素抵抗,探讨隐丹参酮对糖代谢、激素以及卵巢信号分子的影响.方法 40只18 g清洁级昆明白雌性小鼠进行卵巢器官体外培养,加入300 nmol/L地塞米松持续干预48 h,取2组加入隐丹参酮和曲格列酮干预48 h,检测卵巢应用培养液葡萄糖变化情况,利用RT-PCR技术检测AKT2、CSK3β、ERK、MCN2、CYP17、CYP19、AMH基因表达水平.采用t检验进行统计学分析.结果 地塞米松诱导48 h后培养液葡萄糖利用下降,睾酮[分别为(1.85±0.81)、(0.13±0.12)nmol/L,t=4.21,P=0.006]、雄烯二酮[分别为(6.44±0.69)、(0.24±0.15)μg/L,t=-5.11,P=0.002]升高,而孕酮[分别为(14.64±0.68)、(79.38±3.87)nmol/L,t=5.96,P=0.001]、17-羟孕酮[分别为(0.18±0.09)、(0.44±0.06)μg/L,t=3.408,P=0.014]下降.AKT2、GSK3β、ERK mRNA表达下调,MCM2、CYP17表达上调,AMH各组无表达.隐丹参酮干预后,处理组葡萄糖利用升高,睾酮[分别为(0.78±0.23)、(1.85±0.81)nmol/L,t=-2.547,P=0.044]、雄烯二酮[分别为(4.07±0.50)、(6.44±0.69)μg/L,t=4.019,P=0.009]、孕酮[分别为(2.97±1.02)、(14.64±0.68)nmol/L,t=5.959,P=0.001]明显下降,17-羟孕酮[分别为(0.40±0.08)、(0.18±0.09)μg/L,t=-3.113,P=0.036]升高,信号分子表达明显改善.曲格列酮干预结果与之相似,仅孕酮和17-羟孕酮水平变化相反.结论 地塞米松诱导卵巢造成胰岛素抵抗,卵巢增殖功能异常,雄激素合成亢进.隐丹参酮对糖代谢、细胞增殖、激素合成的信号分子具有调控作用.

Objective We hypothesize that cryptotanshinone may directly reduce both insulin resistance and AE of the direct contribution of insulin resistance on ovary induced by dexamethasone. Methods Ovaries from mature mice were isolated and cultured. Insulin resistance was induced in ovaries by dexamethasone (Dex) treatment for 48 h. Insulin-resistant ovaries were further intervened by cryptotanshinone or vericle DMSO in 48 h, and were analyzed with regards to medium glucose, testosterone, androstanedione, progesterone, 17-hydroxide progesterone and estradiol as well as the cellular expression of key signal molecules by RT-PCR. Results Ovaries induced by Dex had a significantly decreased level of glucose utilization, but elevated levels of testosterone and androstanedione as compared with untreated ovaries in medium, indicating establishment of insulin resistance and enhanced androgenic potentials were explored in the induced ovarian. Cryptotanshinone improved glucose utilization in medium, reduced testosterone and androstanedione as compared with vehicle treatment in induced ovaries, suggesting improve significantly insulin resistance and steroidogenesis in induced ovaries. The RT-PCR results showed that dexamethasone decrease the expressions of protein kinase B (AKT2), glycogen synthase kinase-3β (GSK3β) and extracellular regulated protein (ERK), but increased the expressions of cytochrome 17-hydroxylase and minichromosome maitenance protein 2 (MCM2). Cryptotanshinone could improve expression of these signaling molecular mRNA. However, estrogens and its enzymes were reported to be unchanged. Thus, such a differential impacts on theca and granulose by dex will potentiate androgen production in IR ovary. And troglitazone plays similar results as cryptotanshinone except androstanedione, progesterone, 17-hydroxide progesterone. Conclusions Dex could directly induce insulin resistance and androgenic excess within ovaries and crytotanshinon intervene showed beneficial effect. The beneficial effect of cryptotanshinone could ameliorate remarkably molecular regulating proteins of glucose metabolism, cell proliferation and hormone production. This may be a mechanism of Chinese medicine cryptotanshinone improving insulin resistance and androgen synthesis.