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小檗碱和葛根素对猪卵巢膜细胞地塞米松诱导的胰岛素抵抗的作用

[Of berberine and puerarin on dexamethasone-induced insulin resistance in porcine ovarian thecal cells].

作者信息

Gao Lei, Li Wei, Kuang Hong-Ying

机构信息

Department of Obstetrics and Gynecology, First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2009 Jul;29(7):623-7.

Abstract

OBJECTIVE

To explore the direct contribution of dexamethasone (Dex) for insulin resistance inducing in thecal cells and effects of berberine (Ber) and puerarin (Pue).

METHODS

Ovarian thecal cells from porcine follicles were isolated and cultured in vitro. Insulin resistance of thecal cells was induced by Dex treatment for 48 h. Then the glucose utilization ratio of thecal cells was detected. Meanwhile, the effects of Ber and Pue on insulin signal transmission and steroid hormones synthesis were determined by RT-PCR and Western blotting.

RESULTS

(1) After being treated by Dex for 48 h, the [3-3H] -glucose uptake in cells was lowered by about half, and the glucose content in supernate increased for about 1/3. (2) The RT-PCR and Western blotting showed that levels of insulin receptor substrate-1 (IRS-1), mRNA and protein expression of glucose transporter 4 (GLUT4) lowered, peroxisome proliferator-activated receptor -gamma(PPARgamma) and cytochrome P450 17 hydroxylase (CYP17) mRNA or protein expression increased in the model cells. However, the changes of above insulin signal molecules and CYP17 expression were inversed significantly after treated with Ber and Pue for 48 h. (3) As compared with the control, in the model cells, levels of testosterone (T, microg/mL) was higher (0.82 +/- 0.20 vs 0.38 +/- 0.01, P < 0.05), while after Ber and Pue treatment it was 0.44 +/- 0.24 and 0.45 +/- 0.21 respectively, all lower than that in the model cells (P < 0.05). No significant change of serum progesterone was found in all groups (P > 0.05).

CONCLUSIONS

After insulin resistance has been induced, the androgen synthetic capacity of thecal cells enhanced significantly. Ber and Pue could lower the degree of insulin resistance and the androgen synthesis in the model cells, displaying the favorable prospect of the two insulin sensitizing agents for the treatment of polycystic syndrome.

摘要

目的

探讨地塞米松(Dex)对卵泡膜细胞胰岛素抵抗的直接作用以及黄连素(Ber)和葛根素(Pue)的影响。

方法

分离猪卵泡的卵巢卵泡膜细胞并进行体外培养。通过Dex处理48小时诱导卵泡膜细胞产生胰岛素抵抗。然后检测卵泡膜细胞的葡萄糖利用率。同时,采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测Ber和Pue对胰岛素信号转导及甾体激素合成的影响。

结果

(1)经Dex处理48小时后,细胞内[3-3H] -葡萄糖摄取量降低约一半,上清液中葡萄糖含量增加约1/3。(2)RT-PCR和蛋白质免疫印迹法显示,模型细胞中胰岛素受体底物-1(IRS-1)、葡萄糖转运蛋白4(GLUT4)的mRNA和蛋白表达水平降低,过氧化物酶体增殖物激活受体-γ(PPARγ)和细胞色素P450 17羟化酶(CYP17)的mRNA或蛋白表达增加。然而,经Ber和Pue处理48小时后,上述胰岛素信号分子及CYP17表达的变化得到显著逆转。(3)与对照组相比,模型细胞中睾酮(T,μg/mL)水平较高(0.82±0.20 vs 0.38±0.01,P<0.05),而经Ber和Pue处理后分别为0.44±0.24和0.45±0.21,均低于模型细胞(P<0.05)。所有组血清孕酮均无显著变化(P>0.05)。

结论

诱导胰岛素抵抗后,卵泡膜细胞的雄激素合成能力显著增强。Ber和Pue可降低模型细胞的胰岛素抵抗程度及雄激素合成,显示出这两种胰岛素增敏剂治疗多囊综合征的良好前景。

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