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艾塞那肽通过上调PDX-1的表达抑制由高糖及低糖条件诱导的INS-1细胞凋亡的研究
Exenatide can inhibit the apoptosis in INS-1 cell induced by high and low glucose conditions by raising the expression of PDX–1
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DOI:
作者:
陈凌
LING Chen
作者单位:
宁夏医科大学总医院
General Hospital of Ningxia Medical University
关键词:
艾塞那肽对高糖及低糖条件下INS-1细胞PDX-1表达及细胞凋亡的影响
glucagon like peptide -1 analogue ,PDX-1,islet beta cell ,apoptosis
摘要:
【摘要】 目的:探讨GLP-1类似物艾塞那肽在高糖及低糖条件下对胰腺β细胞株INS-1细胞PDX-1表达及细胞凋亡的影响。方法:体外培养INS-1细胞。分为正常对照组NC、 高糖组HG、低糖组DG、高糖艾塞那肽干预组HE、低糖艾塞那肽干预组DE、二甲双胍对照组 HS。相差显微镜观察细胞生长状态;流式细胞仪双染法及原位TUNEL法检测INS-1细胞凋亡情况;分别用Realtime-PCR法、wester blot法及免疫组化法检测胰腺十二指肠同源盒(PDX-1)的表达情况。结果:高糖及低糖环境均可抑制INS-1细胞PDX-1的表达,同时与正常对照组相比高糖组及低糖组凋亡细胞明显增多,凋亡率比较P均<0.05;艾塞那肽干预能明显提高高糖及低糖环境下PDX-1的表达水平,同时细胞凋亡情况得到明显改善,艾塞那肽高糖组与高糖组比较、艾塞那肽低糖组与低糖组比较各个水平PDX-1表达均明显上调,细胞凋亡率明显下降P<0.05;与高糖组比较,二甲双胍高糖干预组细胞凋亡率及PDX-1表达水平无明显变化。结论:在高糖及低糖条件下艾塞那肽均能抑制INS-1细胞的凋亡,同时上调PDX-1的表达水平。
【abstract】 Objective To observe the effect of Exenatide on the expression of PDX-1 and resistance to apoptosis in INS-1 cell Cultured with high and low glucose. Methods The in vitro cultured INS-1 cells were divided into six groups: the control group(NC),high glucose group(HG),low glucose group(DG), high glucose + Exenatied group(HE),low glucose group+ Exenatied group (DG), high glucose + metformin group(HS) , and these cellswere cultured 48 hours respectively. Then we observed the morphological change of cells under the inverted microscope. The apoptosis rate of the INS-1 cells was tested by the flow cytometry and TdT-mediated dUTP nick end labeling. The method of Realtime - PCR, Mr Wester blot method and immunohistochemical method were used to detect the expression of PDX-1 in INS-1 cells. Results Under the high and low glucose the expression of PDX-1 in INS-1 cells were suppressed. In the HG and DG group, the apoptosis rates and apoptosis indexs of the INS-1 cells were higher than the control group, P <0.05 in the comparison of apoptosis rate. After the Exenatide intervention, the expression levels of PDX – 1 in HE and DG group were significantly improved ,apoptosis damage dropped significantly in HE and DE group P <0.05, and there were no significant difference between the HE and DE group about the expression of PDX-1. Compared with the HG group, there were no significant difference about the expression levels of PDX – 1 and the apoptosis rates in HS group. Conclusion Under the environment of High and low Glucose , Exenatide may show resistance to apoptosis of beta cell and up-regulated the expression of PDX-1.
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