高胰岛素对近端肾小管上皮细胞利用β-羟丁酸供能的影响  

Effects of high insulin level on the utilization ofβ-hydroxybutyric acid by proximal renal tubular epithelial cells for adenosine triphosphate production

作　　者：解金兰 常宝成[1] 郭振红 王靖宇[1] 高忠爱 钟菲菲 杨菊红[1] Xie Jinlan;Chang Baocheng;Guo Zhenhong;Wang Jingyu;Gao Zhongai;Zhong Feifei;Yang Juhong(National Health Commission Key Laboratory of Hormones and Development Tianjin Key Laboratory of Metabolic Diseases,Endocrinology Institute of Chu Hsien-I Memorial Hospital and Tianjin Institute of Endocrinology,Tianjin Medical University,Tianjin 300134,China)

机构地区：[1]天津医科大学朱宪彝纪念医院内分泌研究所,天津市内分泌研究所,国家卫健委激素与发育重点实验室,天津市代谢性疾病重点实验室,天津300134

出　　处：《中华糖尿病杂志》2022年第2期179-185,共7页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家重点研发计划(2018YFC1314000);天津市自然科学基金重点项目(17JCZDJC34700)。

摘　　要：目的研究高胰岛素对低糖环境下近端肾小管上皮(HK2)细胞利用β-羟丁酸(BHB)供能的影响及其机制。方法在无糖及低糖环境下,用不同浓度BHB(0、0.5、1.0、2.0、4.0、8.0 mmol/L)干预HK2细胞24 h,检测BHB对HK2细胞增殖能力的影响。在不同时间(0、6、12、24、48 h)下,检测2.0 mmol/L BHB对HK2细胞三磷酸腺苷(ATP)产生的影响。将HK2细胞分为正常对照(NC)组、低糖(LG)组、BHB干预(LG+BHB)组和高胰岛素(LG+BHB+HI)组,检测HK2细胞促凋亡基因Bax mRNA表达水平,抗凋亡基因Bcl2 mRNA、细胞凋亡数量、回吸收白蛋白、ATP产生、线粒体数量以及过氧化物酶体增殖物激活受体共激活因子-1α(PGC1α)、线粒体融合蛋白1(Mfn1)、线粒体动力相关蛋白1(DRP1)蛋白及mRNA表达水平变化。采用不同浓度的胰岛素(5、50 ng/ml)干预HK2细胞,检测Na+偶联单羧酸转运蛋白1(SMCT1)蛋白及mRNA表达水平。两组间比较采用t检验,多组之间比较采用单因素方差分析。结果在无糖环境下,与0 mmol/L BHB组相比,2.0 mmol/L BHB组HK2细胞增殖能力增加(P<0.05);在低糖环境下,与0 mmol/L BHB组相比,2.0 mmol/L BHB组细胞增殖能力无明显变化(P>0.05)。与2.0 mmol/L BHB干预0 h组相比,2.0 mmol/L BHB干预24 h组HK2细胞ATP产生明显增加(P<0.05)。与NC组相比,LG组HK2细胞Bax mRNA表达、细胞凋亡数量、DRP1 mRNA表达均增加(P<0.05),Bcl2 mRNA表达水平、细胞回吸收白蛋白、ATP产生、PGC1α、Mfn1蛋白表达均降低(P<0.05);与LG组相比,LG+BHB组HK2细胞Bax mRNA、细胞凋亡数量、DRP1 mRNA表达均降低(P<0.05),Bcl2 mRNA表达、细胞回吸收白蛋白、ATP产生、PGC1α、Mfn1 mRNA及蛋白表达均增加(P<0.05);与LG+BHB组相比,LG+BHB+HI组HK2细胞Bax mRNA、细胞凋亡数量和DRP1 mRNA表达均增加(P<0.05),Bcl2 mRNA、细胞回吸收白蛋白、ATP产生、PGC1α、Mfn1 mRNA及蛋白表达均降低(P<0.05)。与5 ng/ml胰岛素干预组相比,50 ng/ml胰岛素组细胞SMCT1蛋白及mRNA表Objective To study the effect of high insulin on the energy utilization ofβ-hydroxybutyric acid(BHB)in proximal renal tubular epithelial cells(HK2)under low glucose condition and its related mechanism.Methods HK2 cells were treated with different BHB concentrations(0,0.5,1.0,2.0,4.0,8.0 mmol/L)for 24 h in glucose-free and low glucose environments,to study the effect of BHB on the proliferation of HK2 cells.The effect of 2.0 mmol/L BHB on ATP production of HK2 cells was studied at different time points(0,6,12,24,48 h).HK2 cells were divided into normal control(NC)group,low glucose(LG)group,BHB intervention(LG+BHB)group and high insulin(LG+BHB+HI)group.These various groups were used to study the expression of pro-apoptotic gene Bax mRNA in HK2 cells,anti-apoptotic gene Bcl2 mRNA,apoptosis number,albumin absorption,ATP production,mitochondrial number,peroxisome proliferator activated receptor co-activator-1α(PGC1α),mitochondrial mitofusin 1(Mfn1),mitochondrial dynamin related protein 1(DRP1)protein and mRNA levels.HK2 cells were treated with different insulin(5 ng/ml,50 ng/ml)to study the effects of high insulin on the expression of sodium-coupled monocarboxylate transporter 1(SMCT1)protein and mRNA.The t test was used for comparison between two groups,one-way analysis of variance(ANOVA)was used for comparison between multiple groups.Results In glucose-free environment,compared with 0 mmol/L BHB group,the proliferation of HK2 cells in 2.0 mmol/L BHB group increased(P<0.05).Compared with 0 mmol/L BHB group,there was no significant difference in cell proliferation in 2.0 mmol/L BHB group under low glucose environment(P>0.05).Compared with 2.0 mmol/L BHB intervention group for 0 h,ATP production of HK2 cells in 2.0 mmol/L BHB intervention group for 24 h was significantly increased(P<0.05).Compared with NC group,Bax mRNA,apoptosis number and DRP1 mRNA of HK2 cells in LG group were increased(all P<0.05),Bcl2 mRNA expression,albumin absorption,ATP production,PGC1αand Mfn1 protein expression were decreased(all P<0.05).

关 键 词：高胰岛素血症 Β-羟丁酸 线粒体 近端肾小管上皮细胞 

分 类 号：R692[医药卫生—泌尿科学]