SCFAs调节线粒体自噬改善高糖下胰岛β细胞损伤的研究  

Regulation of Mitophagy by SCFAs Improves the Injury of Isletβ-cell under High Glucose

作　　者：王思瑶[1] 陆春晖[1] 张洁[1] 靳瑾[1] WANG Siyao;LU Chunhui;ZHANG Jie(Department of Endocrinology,The Fifth Affiliated Hospital of Xinjiang Medical University,Xinjiang 830000,China)

机构地区：[1]新疆医科大学第五附属医院内分泌科,乌鲁木齐830000

出　　处：《医学研究杂志》2023年第9期85-91,共7页Journal of Medical Research

基　　金：新疆维吾尔自治区自然科学基金资助项目(2022D01C318)。

摘　　要：目的探究短链脂肪酸(short-chain fatty acids,SCFAs)通过调节线粒体自噬途径改善高糖环境下胰岛β细胞功能损伤的作用及机制。方法将大鼠胰岛素瘤INS-1细胞随机分为对照组、模型组、乙酸钠(sodium acetate,NaA)组、丙酸钠(sodium propionate,NaP)组、丁酸钠(sodium butyrate,NaB)组,除对照组外,其余组均采用33mmol/L葡萄糖培养诱导高糖损伤,NaA组、NaP组及NaB组预先分别以100μmol/L的NaA、NaP、NaB进行处理;各组INS-1细胞处理结束后,CCK-8法检测细胞增殖活性,流式细胞术检测细胞凋亡水平,Hoechst 33258染色观察细胞凋亡形态,放射免疫法测定细胞释放胰岛素的含量,DCFH-DA荧光探针测定细胞活性氧(reactive oxygen species,ROS)水平,透射电子显微镜观察线粒体超微结构及自噬情况,Western blot法检测微管相关蛋白Ⅰ轻链3(microtubule-associated proteinⅠlight 3,LC3-Ⅰ)、LC3-Ⅱ、p62、PINK1及Parkin的表达水平。结果与模型组比较,NaA组、NaP组及NaB组的细胞增殖率升高,而细胞凋亡率下降,未见明显蓝色凋亡体,胰岛素释放量升高,ROS水平减少,线粒体受损与自噬现象得到改善,细胞中LC3-Ⅱ/LC3-Ⅰ比值降低,PINK1与Parkin蛋白相对表达量下调,且p62蛋白相对表达量上调,差异均有统计学意义(P<0.05)。结论3种SCFAs(NaA、NaP、NaB)均能改善高糖环境下胰岛β细胞的功能损伤,其机制可能与抑制线粒体自噬相关。Objective To explore the effect and mechanism of short-chain fatty acids(SCFAs)in improving pancreaticβ-cell dysfunction under high glucose environment by regulating mitophagy pathway.Methods Rat insulinoma INS-1 cells were randomly divided into control group,model group,sodium acetate(NaA)group,sodium propionate(NaP)group,sodium butyrate(NaB)group,except the control group,the other groups were cultured with 33mmol/L glucose to induce high glucose injury,the NaA group,NaP group and NaB group were pretreated with 100μmol/L NaA,NaP and NaB,respectively;after treatment with INS-1 cells in each group,CCK-8 assay was used to detect the cell proliferation activity,flow cytometry was used to detect the level of apoptosis,Hoechst 33258staining was used to observe the morphology of apoptosis,the amount of insulin released by cells was measured by radioimmunoassay,DCFH-DA fluorescent probe was used to measure cellular reactive oxygen species(ROS)levels,transmission electron microscopy was used to observe mitochondrial ultrastructure and autophagy,Western blot was used to detect the expression levels of microtubule-associated proteinⅠlight chain 3(LC3-Ⅰ),LC3-Ⅱ,p62,PINK1 and Parkin.Results Compared with the model group,the proliferation rate of cells in the NaA group,NaP group and NaB group was increased,while the apoptosis rate of cells was decreased,no obvious blue apoptotic bodies was found,the release of insulin was increased,the level of ROS was decreased,mitochondrial damage and autophagy were improved,the ratio of LC3-Ⅱ/LC3-Ⅰwas decreased,the relative protein expression of PINK1 and Parkin were down-regulated,and the relative protein expression of p62 was up-regulated,the differences were statistically significant(P<0.05).Conclusion Three SCFAs(NaA,NaP,NaB)can improve the dysfunction of isletβ-cell under high glucose environment,and the mechanism may be related to the inhibition of mitophagy.

关 键 词：短链脂肪酸 胰岛Β细胞 高糖 线粒体自噬 

分 类 号：R587.2[医药卫生—内分泌]