| 引用本文:莫芳芳1，刘海霞1，华静2，赵丹丹1，高思华1.降糖消渴颗粒含药血清对INS-1细胞磷酸肌醇-3激酶/AKT/FOXO1通路的影响[J].世界中医药,2019,(02):. |
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| 降糖消渴颗粒含药血清对INS-1细胞磷酸肌醇-3激酶/AKT/FOXO1通路的影响 |
| Effects of Jiangtang Xiaoke Granules Containing Serum on Phosphoinositide-3 Kinase/AKT/FOXO1 Signaling Pathway in INS-1 Cells |
| 投稿时间：2018-03-09 |
| DOI：10.3969/j.issn.1673-7202.2019.02.020 |
| 中文关键词: 肝脾肾同调 降糖消渴颗粒 PI3K/AKT/FOXO1通路 FOXO1因子 胰腺 INS-1细胞 2型糖尿病 含药血清 |
| English Keywords:Common regulation of liver, spleen and kidney Jiangtang Xiaoke Granules, PI3K/AKT/FOXO1 signaling pathway FOXO1 pancreas INS-1 cells Type 2 diabetes mellitus Serum containing drug |
| 基金项目:国家自然科学基金青年科学基金项目（NSFC81703971）;校内课题资助项目（2016-ZXFZJJ-109） |
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| 中文摘要: |
| 目的：通过研究降糖消渴颗粒含药血清对INS-1细胞PI3K/AKT/FOXO1通路中相关蛋白与基因表达的影响，探讨降糖消渴颗粒含药血清保护胰岛B细胞的分子机制。方法：选取FOXO1高表达INS-1稳定细胞株，分别以1%、5%、10%、15%、20%不同浓度降糖消渴颗粒含药血清干预，用MTT法计算细胞成活率以观察药物毒性，分别检测各组细胞中总FOXO1蛋白水平以确定最佳药物浓度开展后续实验。细胞用或不用PI3K抑制剂LY294002干预后，以Western blotting检测细胞中总FOXO1、p-FOXO1、Akt、p-Akt和细胞核内外的FOXO1、p-FOXO1的蛋白表达，qRT-PCR检测FOXO1mRNA和AktmRNA含量。结果：不同浓度含药血清对细胞均无毒性，其中10%为最佳药物干预浓度。10%含药血清使磷酸化FOXO1和Akt的表达升高；在胞质和胞核中，10%含药血清使FOXO1的表达降低，磷酸化表达升高，抑制剂LY294002使FOXO1和p-FOXO1表达均降低。实时荧光定量PCR（qRT-PCR）结果显示，Akt mRNA表达升高，FOXO1mRNA表达降低，抑制剂使FOXO1mRNA表达降低，结果与Western blotting结果一致。结论：降糖消渴颗粒含药血清可通过PI3K/AKT/FOXO1通路促进FOXO1磷酸化出核以抑制FOXO核转录实现保护胰岛B细胞的作用。 |
| English Summary: |
| To study the effects of Jiangtang Xiaoke Granules containing serum on expressions of protein and genes of phosphoinositide-3 kinase (PI3K)/AKT/FOXO1 signaling pathway in INS-1 cells to explore the molecular mechanism of serum contained Jiangtang Xiaoke Granules on protecting β-cells. Methods:The FOXO1 high expression INS-1 stable cell line was selected and treated with 1%, 5%, 10%, 15% and 20% different concentrations of Jiangtang Xiaoke Granules containing serum. The cell survival rate was calculated by MTT method to observe the toxicity of the drug. The total FOXO1 protein level in each group was detected to determine the optimal drug concentration for subsequent experiments. The cells were transfected with or without the PI3K inhibitor LY294002. Western blotting was used to detect the expression of FOXO1, p-FOXO1, Akt, p-Akt and FOXO1 and p-FOXO1 in the nucleus. The expression of FOXO1 mRNA and Akt mRNA was detected by qRT-PCR. Results:Different concentrations of drug-containing serum were not toxic to cells, and 10% of them were the optimal drug intervention concentration. The expression of phosphorylated FOXO1 and Akt was increased with 10% drug-containing serum. In the cytoplasm and nucleus, the expression of FOXO1 was decreased and the expression of phosphorylation was increased with 10% drug-containing serum, and the expression of FOXO1 and p-FOXO1 were decreased by the inhibitor LY294002. The results of real-time quantitative PCR (qRT-PCR) showed that the expression of Akt mRNA was increased, the expression of FOXO1 mRNA was decreased, and the expression of FOXO1 mRNA was decreased by inhibitor. The results were consistent with the results of Western blotting. Conclusion:The drug-containing serum of Jiangtang Xiaoke Granules can promote the phosphorylation of FOXO1 through the PI3K/AKT/FOXO1 pathway to inhibit the transcription of FOXO nuclear to protect islet B cells. |
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