高糖高脂环境下生长激素释放肽对胰腺β细胞自噬的影响（英文）  

Effect of ghrelin on autophagy of pancreatic β cells under high glucose and high lipid environment

作　　者：袁雪 廖婷[1] 荣曦[1] 刘红[1] Xue Yuan;Ting Liao;Xi Rong;Hong Liu

机构地区：[1]广西医科大学第一附属医院老年内分泌代谢科,南宁530021

出　　处：《广西医科大学学报》2019年第4期495-500,共6页Journal of Guangxi Medical University

基　　金：supported by grants from the National Natural Science Foundation of China(No.81760154,No.31600677);Guangxi Natural Science Foundation ( No. 2017JJA10168);Guangxi University Student Innovation and Entrepreneurship Training Program (No.201710598006,No.20180598030)

摘　　要：目的:探讨高糖高脂(HGHL)环境下生长激素释放肽(ghrelin)对胰腺β细胞自噬的影响。方法:将胰岛NIT-1细胞分为对照组(葡萄糖5.6mmol/L)、ghrelin组(葡萄糖5.6mmol/L+100nmol/L ghrelin)、HGHL组(葡萄糖33.3mmol/L+0.5mol/L棕榈酸)、HGHL+ghrelin组(葡萄糖33.3mmol/L+0.5mol/L棕榈酸+100nmol/L ghrelin)。采用CCK-8法检测NIT-1细胞活性。采用单丹磺酰戊二胺(MDC)染色和微管相关蛋白1轻链3(LC3)B免疫荧光染色检测细胞自噬活性。结果:HGHL+ghrelin组细胞活力、Beclin 1蛋白水平、LC3Ⅱ/LC3Ⅰ的比值、Atg6和LC3B基因相对表达量均显著高于HGHL组(均P<0.05)。MDC染色和免疫荧光染色显示,HGHL+ghrelin组细胞内自噬泡数量和阳性LC3B斑点数目增加。结论:Ghrelin能保护胰岛β细胞免受HGHL损伤,可能与自噬能力的增强有关。Objective: To investigate the effect of ghrelin on the autophagy of islet β cells under high glucose and high lipid (HGHL) environment. Methods: Islet NIT-1 cells were cultured and divided into four groups: control group(glucose 5.6 mmol/L),ghrelin group(glucose 5.6 mmol/L+100 nmol/L ghrelin),HGHL group (glucose 33.3 mmol/L+0.5 mmol/L palmitic acid),HGHL+ghrelin group(glucose 33.3 mmol/L+0.5 mmol/L PA+100 nmol/L ghrelin). The cell counting kit (CCK-8) was used to assess the NIT-1 cells viability. Monodansylcadaverine(MDC)staining and microtubule-associated protein 1 light chain 3 (LC3) B immunofluorescence staining were employed to detect the autophagy. Western blotting was performed to determine the protein expression of Beclin 1 and the ratio of LC3Ⅱto LC3Ⅰ,and real-time fluorescence quantitative PCR (qPCR)was tomeasure the mRNA expression of Atg6 and LC3B. Results: The cell viability, the protein level of Beclin 1, the ratio of LC3Ⅱ/LC3Ⅰ, and the mRNA expression levels of Atg6 and LC3B in HGHL+ghrelin groupweresignificantly increased compared with HGHL group (P<0.05). MDC staining and immunofluorescence staining also showed increased deposition of autophagic vesicles and autophagic protein LC3B in HGHL+ghrelin group. Conclusion: Ghrelin could protect islet β cells from HGHL injury,which might be related to the enhancement of autophagy.

关 键 词：GHRELIN 胰岛β细胞 自噬 高脂高糖 

分 类 号：R392.114[医药卫生—免疫学]