中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2012年
6期
357-360
,共4页
柳彬彬%宋璐璐%萧建中%李世蕊%杨文英
柳彬彬%宋璐璐%蕭建中%李世蕊%楊文英
류빈빈%송로로%소건중%리세예%양문영
利拉鲁肽%胰岛β细胞%微RNAs
利拉魯肽%胰島β細胞%微RNAs
리랍로태%이도β세포%미RNAs
Liratglutide%Islet beta-cells%MicroRNAs
目的 探讨利拉鲁肽对db/db小鼠胰腺及大鼠胰岛细胞瘤细胞系INS-I中microRNA表达的影响.方法 将20只4周龄雄性db/db小鼠按随机数字表法分为对照组和利拉鲁肽组(每组10只).分别给予0.1ml生理盐水或300 ng/g利拉鲁肽皮下注射,每日2次.8周后测定糖化血红蛋白,行腹腔注射葡萄糖耐量试验(TPGTT).8周末处死小鼠,免疫组化法分析小鼠胰腺β细胞增殖水平；实时荧光定量聚合酶链反应( RT-PCR)测定小鼠胰腺microRNA(miR-375和miR-34a)的表达.INS-1细胞分别给予0.5 mmol/L棕榈酸培养0、48、72 h或100 nmol/L利拉鲁肽预处理12h后,给予0.5 mmol/L棕榈酸培养72 h,RT-PCR测定miR-375和miR-34a表达水平.采用t检验及方差分析进行组间数据比较分析.结果 与对照组相比,利拉鲁肽组小鼠糖化血红蛋白水平降低(分别为7.3％±0.3％和4.7％±0.6％,t=16.47,P＜0.01)；IPGTT血糖曲线下面积降低[分别为( 4568±197)和(1927±127) mmol·L-1·min-1,t =26.53,P＜0.05]；胰岛素曲线下面积增加[分别为(1080±247)和(2818±378)μg·L-1·min-1,t=7.73,P＜0.05].免疫组化法显示,与对照组相比,利拉鲁肽组小鼠胰岛素阳性面积增加(分别为1.40±0.30和0.37±0.09,t=19.14,P＜0.01),Brdu染色阳性细胞比例增加(分别为2.40％±0.22％和0.73％±0.10％,t=4.97,P＜0.01).利拉鲁肽组小鼠胰腺miR-375与miR-34a表达较对照组分别降低50％(分别为1.1±0.3和2.2±0.5,t=3.08,P＜0.05)和71％(分别为1.1±0.3和3.8±1.2,t=2.80,P＜0.05).棕榈酸培养使INS-1细胞miR-375表达呈剂量、时间依赖性增加,利拉鲁肽可抑制棕榈酸诱导的miR-375表达(F=7.20,P＜0.01).结论 microRNA可能是利拉鲁肽调节β细胞增殖的靶点之一.
目的 探討利拉魯肽對db/db小鼠胰腺及大鼠胰島細胞瘤細胞繫INS-I中microRNA錶達的影響.方法 將20隻4週齡雄性db/db小鼠按隨機數字錶法分為對照組和利拉魯肽組(每組10隻).分彆給予0.1ml生理鹽水或300 ng/g利拉魯肽皮下註射,每日2次.8週後測定糖化血紅蛋白,行腹腔註射葡萄糖耐量試驗(TPGTT).8週末處死小鼠,免疫組化法分析小鼠胰腺β細胞增殖水平；實時熒光定量聚閤酶鏈反應( RT-PCR)測定小鼠胰腺microRNA(miR-375和miR-34a)的錶達.INS-1細胞分彆給予0.5 mmol/L棕櫚痠培養0、48、72 h或100 nmol/L利拉魯肽預處理12h後,給予0.5 mmol/L棕櫚痠培養72 h,RT-PCR測定miR-375和miR-34a錶達水平.採用t檢驗及方差分析進行組間數據比較分析.結果 與對照組相比,利拉魯肽組小鼠糖化血紅蛋白水平降低(分彆為7.3％±0.3％和4.7％±0.6％,t=16.47,P＜0.01)；IPGTT血糖麯線下麵積降低[分彆為( 4568±197)和(1927±127) mmol·L-1·min-1,t =26.53,P＜0.05]；胰島素麯線下麵積增加[分彆為(1080±247)和(2818±378)μg·L-1·min-1,t=7.73,P＜0.05].免疫組化法顯示,與對照組相比,利拉魯肽組小鼠胰島素暘性麵積增加(分彆為1.40±0.30和0.37±0.09,t=19.14,P＜0.01),Brdu染色暘性細胞比例增加(分彆為2.40％±0.22％和0.73％±0.10％,t=4.97,P＜0.01).利拉魯肽組小鼠胰腺miR-375與miR-34a錶達較對照組分彆降低50％(分彆為1.1±0.3和2.2±0.5,t=3.08,P＜0.05)和71％(分彆為1.1±0.3和3.8±1.2,t=2.80,P＜0.05).棕櫚痠培養使INS-1細胞miR-375錶達呈劑量、時間依賴性增加,利拉魯肽可抑製棕櫚痠誘導的miR-375錶達(F=7.20,P＜0.01).結論 microRNA可能是利拉魯肽調節β細胞增殖的靶點之一.
목적 탐토리랍로태대db/db소서이선급대서이도세포류세포계INS-I중microRNA표체적영향.방법 장20지4주령웅성db/db소서안수궤수자표법분위대조조화리랍로태조(매조10지).분별급여0.1ml생리염수혹300 ng/g리랍로태피하주사,매일2차.8주후측정당화혈홍단백,행복강주사포도당내량시험(TPGTT).8주말처사소서,면역조화법분석소서이선β세포증식수평；실시형광정량취합매련반응( RT-PCR)측정소서이선microRNA(miR-375화miR-34a)적표체.INS-1세포분별급여0.5 mmol/L종려산배양0、48、72 h혹100 nmol/L리랍로태예처리12h후,급여0.5 mmol/L종려산배양72 h,RT-PCR측정miR-375화miR-34a표체수평.채용t검험급방차분석진행조간수거비교분석.결과 여대조조상비,리랍로태조소서당화혈홍단백수평강저(분별위7.3％±0.3％화4.7％±0.6％,t=16.47,P＜0.01)；IPGTT혈당곡선하면적강저[분별위( 4568±197)화(1927±127) mmol·L-1·min-1,t =26.53,P＜0.05]；이도소곡선하면적증가[분별위(1080±247)화(2818±378)μg·L-1·min-1,t=7.73,P＜0.05].면역조화법현시,여대조조상비,리랍로태조소서이도소양성면적증가(분별위1.40±0.30화0.37±0.09,t=19.14,P＜0.01),Brdu염색양성세포비례증가(분별위2.40％±0.22％화0.73％±0.10％,t=4.97,P＜0.01).리랍로태조소서이선miR-375여miR-34a표체교대조조분별강저50％(분별위1.1±0.3화2.2±0.5,t=3.08,P＜0.05)화71％(분별위1.1±0.3화3.8±1.2,t=2.80,P＜0.05).종려산배양사INS-1세포miR-375표체정제량、시간의뢰성증가,리랍로태가억제종려산유도적miR-375표체(F=7.20,P＜0.01).결론 microRNA가능시리랍로태조절β세포증식적파점지일.
Objective To investigate the effect of glucagon-like peptide 1 analogue liraglutide on microRNA expression of db/db mice and rat insulinoma cell line( INS-l).Methods Twenty 4-week old male db/db mice were randomly assigned to receive a subcutaneous injection of liraglutide(300 ng/g bid) or normal saline (0.1 ml bid) ( control group) according to random numbers table.Glycated hemoglobin (HbAlc) was determined before and after 8 weeks of intervention.At the end of the intervention,intraperitoneal glucose tolerance test ( IPGTT) was performed Beta cells proliferation rate was determined by using immunohistochemistry methods.INS-1 cells were cultured with 0.5 mmol/L palmitic acid for 0.48,72 h or treated by 100 nmol/L liraglutide for 12 h before 72 h of palmitic acid ( 0.5 mmol/L) culture.microRNA miR-375 and miR-34a expression were examined by real-time polymerase chain reaction ( RTPCR).The t test or ANOVA analysis was used for data analysis.Results Compared with the control group,HbAle in the liraglutide group was significantly lower ( 7.3％ ± 0.3％ vs 4.7％ ± 0.6％,t =16.47,P ＜ 0.01); the glucose aera under curve in the liraglutide group was decreased significantly (( 4568 ± 197) vs ( 1927 ± 127) mmol · L-1 · min -1,t =26.53,P ＜ 0.05 ),while insulin aera under curve was increased significantly ( ( 1080 ± 247 ) vs ( 2818 4± 378 ) μg· L- 1.min - 1,t =7.73,P ＜ 0.05 ).Immunohistochemistry showed more insulin staining in single pancreas of db/db mice after liraglutide treatment than in the control group ( 1.40 ±0.30 vs 0.37 ±0.09,t =19.14,P <0.01 ).More Brdu-positive cells were observed in islets of liraglutide-treated mice than in control mice after intervention ( 2.40％ ±0.22％ vs 0.73％ ±0.10％,t =4.97,P ＜0.01 ).Compared with control mice,the miR-375 expression of pancreas in liraglutide-treated db/db mice was reduced by 50％ ( 1.1 ± 0.3 vs 2.2 ± 0.5,t =3.08,P ＜0.05 ) and miR-34a was reduced by 71％ ( 1.1 ± 0.3 vs 3.8 ± 1.2,t =2.80,P ＜ 0.05 ).In addition,the miR-375 expression of INS-1 cells was up-regulated dose- and time-dependently in response to palmitic acid;and palmitic acid-induced miR-375 expression was suppressed by liraglutide in vitro( F =7.20,P ＜ 0.01 ).Conclusion Liraglutide may preserve beta-cell proliferation in response to lipotoxicity through a regulation of miRNA.
