HtrA 1基因敲除对小鼠葡萄糖代谢及胰岛功能的影响  

Effect of HtrA1 gene knockout on glucose metabolism and pancreatic cell function in mice

作　　者：高新雨 倪嘉成 白宁宁[1] 张菁[1] 高琼媚 李华婷[1] 杨颖 方启晨[1] GAO Xinyu;NI Jiacheng;BAI Ningning;ZHANG Jing;GAO Qiongmei;LI Huating;YANG Ying;FANG Qichen(Shanghai Diabetes Institute,Shanghai Key Laboratory of Diabetes Mellitus,Shanghai Clinical Center for Diabetes,Shanghai Clinical Center for Metabolic Disease,Department of Endocrinology,Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 200233,China)

机构地区：[1]上海交通大学附属第六人民医院内分泌代谢科,上海市糖尿病研究所,上海市糖尿病重点实验室,上海市糖尿病临床医学中心,上海市代谢病临床医学中心,上海200233

出　　处：《上海医学》2022年第6期394-399,共6页Shanghai Medical Journal

基　　金：上海市自然科学基金(15ZR1431700)。

摘　　要：目的探讨HtrA 1基因敲除对小鼠糖代谢及胰岛功能的影响。方法选取6周龄雄性野生型C57BL/6J小鼠(WT小鼠)和HtrA 1基因敲除小鼠(KO小鼠)按照同窝随机对照原则分组,分别给予高脂饮食(脂肪占总热量的60%)或普脂饮食(脂肪占总热量的5%),记录喂养16周时两组WT小鼠、KO小鼠的体重及喂养期间摄食量。对18周龄(普脂或高脂喂养12周)小鼠行葡萄糖耐量试验(IPGTT),并测定空腹胰岛素水平,进一步对21周龄(高脂喂养15周)小鼠行葡萄糖刺激胰岛素分泌(GSIS)试验,观察糖代谢及胰岛素分泌情况。分离小鼠胰岛在细胞水平行GSIS试验,测定胰岛素水平。取22周龄(高脂喂养16周)小鼠胰腺组织行组织切片,进行胰岛素(绿色)和胰高血糖素(红色)免疫荧光双染,以及胰岛素(绿色)和CD31(红色)免疫荧光双染。应用ZEN 2.0软件对胰岛中CD31水平进行定量,比较WT小鼠与KO小鼠胰岛中CD31的平均荧光强度。结果高脂饮食组或普脂饮食组的KO与WT小鼠的体重和摄食量的差异均无统计学意义(P值均>0.05)。普脂饮食组KO小鼠糖负荷10、20、120 min时的血糖水平均显著高于同组WT小鼠(P值均<0.05),空腹血糖及糖负荷后30、45、60、90 min时的血糖水平与同组WT小鼠的差异均无统计学意义(P值均>0.05)。高脂饮食组KO小鼠糖负荷10、20、30、45 min时血糖水平均显著高于同组WT小鼠(P值均<0.05),空腹血糖及糖负荷后60、90、120 min时血糖水平与同组WT小鼠的差异均无统计学意义(P值均>0.05)。18周龄普脂饮食组WT小鼠与KO小鼠空腹胰岛素水平的差异无统计学意义(P>0.05),高脂饮食组KO小鼠空腹胰岛素水平显著低于WT小鼠(P<0.05)。21周龄高脂饮食组空腹及注射葡萄糖后15 min时KO小鼠的胰岛素水平均显著低于WT小鼠(P值均<0.05),KO小鼠与WT小鼠注射葡萄糖后30 min时胰岛素水平的差异无统计学意义(P>0.05)。经高糖刺激(20 mmol/L葡萄糖处理)的KO小鼠胰Objective To explore the effect of HtrA1 knockout on glucose metabolism and pancreatic cell function in mice.Methods Six-week-old male wild-type C57BL/6J mice and HtrA1 knockout mice were randomly divided into high-fat diet group(60%kcal from fat)and normal-chow diet group(5%kcal from fat).The body weight and food intake of wild-type and gene knockout mice in both groups were recorded at 16 weeks of feeding.Intrapertoneal glucose tolerance testing(IPGTT)and fasting insulin level were assessed on 18-week-old mice(12 weeks of mormal-chow or high-fat feeding).The plasma insulin secretion was assessed by glucose-stimulated insulin secretion(GSIS)assay in 21-week-old mice(15 weeks of high-fat feeding).The level of plasma insulin was determined in isolated islets by a GSIS assay.Immunofluorescence staining for insulin(green),glucagon(red)and CD31(red)was performed on pancreatic tissue from 22-week-old mice(16 weeks of high-fat feeding).The expression of CD31 in islets was quantified by ZEN 2.0 software,and the mean fluorescence intensity of CD31 in islets was compared between wild-type and gene knockout mice.Results There was no significant difference in body weight or food intake between wild-type and gene knockout mice with high-fat diet or normal-chow diet(both P>0.05).In the normal-chow diet group,the blood glucose level in gene knockout mice was significantly higher than that in wild-type mice at 10,20 and 120 min after glucose injection(all P<0.05),but there was no significant difference in the blood glucose level at 0,30,45,60 and 90 min after glucose injection between wild-type and gene knockout mice(all P>0.05).In the high-fat diet group,the blood glucose level in gene knockout mice was significantly higher than that in wild-type mice at 10,20,30 and 45 min after glucose injection(all P<0.05),but there was no significant difference in the blood glucose level at 0,60,90 and 120 min after glucose injection between wild-type and gene knockout mice(all P>0.05).In the normal-chow diet group,there was no significant

关 键 词：HtrA 1基因 葡萄糖代谢 胰岛细胞 

分 类 号：R587.1[医药卫生—内分泌]