南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2015年
5期
671-676
,共6页
易秋艳%刘艳清%张珍%刘春燕%卢斌%邵加庆
易鞦豔%劉豔清%張珍%劉春燕%盧斌%邵加慶
역추염%류염청%장진%류춘연%로빈%소가경
RNA干扰%肾素-血管紧张素系统%血管紧张素II 1型受体%第一相胰岛素分泌
RNA榦擾%腎素-血管緊張素繫統%血管緊張素II 1型受體%第一相胰島素分泌
RNA간우%신소-혈관긴장소계통%혈관긴장소II 1형수체%제일상이도소분비
RNA interference%rennin-angiotensin system%angiotensin II type 1 receptor%first-phase insulin secretion
目的:观察RNA干扰技术阻断胰岛局部血管紧张素II 1型受体(AT1R)表达后db/db小鼠胰岛第一相胰岛素分泌的变化并探讨其潜在机制。方法分离db/db和db/m小鼠的胰岛并检测AT1R mRNA和蛋白的表达。构建针对小鼠AT1R基因的RNA干扰重组腺病毒(Ad-siAT1R)及含对照序列的重组腺病毒(Ad-siControl)。将分离培养的db/db小鼠胰岛细胞分为三组:Ad-siAT1R感染组、Ad-siControl感染组、空白对照组。腺病毒感染后继续培养胰岛细胞72 h。检测各组AT1R、GLUT-2及葡萄糖激酶(GCK)的表达,并用胰岛灌流系统检测胰岛素动态分泌。结果 db/db小鼠胰岛中AT1R mRNA和蛋白表达水平比db/m小鼠胰岛高2倍左右(P<0.05)。腺病毒感染后,Ad-siAT1R组较Ad-siControl组胰岛AT1R mRNA表达水平下降75%,蛋白表达水平下降65%,而GLUT-2及GCK表达水平分别升高190%、121%(均P<0.05)。胰岛灌流显示:空白对照组和Ad-siControl组小鼠的胰岛素第一相分泌显著下降,仅为基础水平的1.8倍;而Ad-siAT1R组在高糖负荷后1~2 min即达到最高峰值140 mU/L,为基础水平的2.8倍,表明第一相胰岛素分泌明显改善。结论 RNA干扰特异性阻断胰岛局部AT1R表达可上调GLUT-2及GCK表达,恢复第一相胰岛素分泌,这可能是AT1R阻滞剂改善胰岛分泌功能的机制之一。
目的:觀察RNA榦擾技術阻斷胰島跼部血管緊張素II 1型受體(AT1R)錶達後db/db小鼠胰島第一相胰島素分泌的變化併探討其潛在機製。方法分離db/db和db/m小鼠的胰島併檢測AT1R mRNA和蛋白的錶達。構建針對小鼠AT1R基因的RNA榦擾重組腺病毒(Ad-siAT1R)及含對照序列的重組腺病毒(Ad-siControl)。將分離培養的db/db小鼠胰島細胞分為三組:Ad-siAT1R感染組、Ad-siControl感染組、空白對照組。腺病毒感染後繼續培養胰島細胞72 h。檢測各組AT1R、GLUT-2及葡萄糖激酶(GCK)的錶達,併用胰島灌流繫統檢測胰島素動態分泌。結果 db/db小鼠胰島中AT1R mRNA和蛋白錶達水平比db/m小鼠胰島高2倍左右(P<0.05)。腺病毒感染後,Ad-siAT1R組較Ad-siControl組胰島AT1R mRNA錶達水平下降75%,蛋白錶達水平下降65%,而GLUT-2及GCK錶達水平分彆升高190%、121%(均P<0.05)。胰島灌流顯示:空白對照組和Ad-siControl組小鼠的胰島素第一相分泌顯著下降,僅為基礎水平的1.8倍;而Ad-siAT1R組在高糖負荷後1~2 min即達到最高峰值140 mU/L,為基礎水平的2.8倍,錶明第一相胰島素分泌明顯改善。結論 RNA榦擾特異性阻斷胰島跼部AT1R錶達可上調GLUT-2及GCK錶達,恢複第一相胰島素分泌,這可能是AT1R阻滯劑改善胰島分泌功能的機製之一。
목적:관찰RNA간우기술조단이도국부혈관긴장소II 1형수체(AT1R)표체후db/db소서이도제일상이도소분비적변화병탐토기잠재궤제。방법분리db/db화db/m소서적이도병검측AT1R mRNA화단백적표체。구건침대소서AT1R기인적RNA간우중조선병독(Ad-siAT1R)급함대조서렬적중조선병독(Ad-siControl)。장분리배양적db/db소서이도세포분위삼조:Ad-siAT1R감염조、Ad-siControl감염조、공백대조조。선병독감염후계속배양이도세포72 h。검측각조AT1R、GLUT-2급포도당격매(GCK)적표체,병용이도관류계통검측이도소동태분비。결과 db/db소서이도중AT1R mRNA화단백표체수평비db/m소서이도고2배좌우(P<0.05)。선병독감염후,Ad-siAT1R조교Ad-siControl조이도AT1R mRNA표체수평하강75%,단백표체수평하강65%,이GLUT-2급GCK표체수평분별승고190%、121%(균P<0.05)。이도관류현시:공백대조조화Ad-siControl조소서적이도소제일상분비현저하강,부위기출수평적1.8배;이Ad-siAT1R조재고당부하후1~2 min즉체도최고봉치140 mU/L,위기출수평적2.8배,표명제일상이도소분비명현개선。결론 RNA간우특이성조단이도국부AT1R표체가상조GLUT-2급GCK표체,회복제일상이도소분비,저가능시AT1R조체제개선이도분비공능적궤제지일。
Objective To investigate the effects of angiotensin II type 1 receptor (AT1R) knockdown on the first-phase insulin secretion in isolated islets of db/db mice and explore the possible mechanisms. Methods Islets were isolated from db/db and db/m mice and the expression level of AT1R in the islets was assayed. A recombinant adenovirus containing siRNA targeting AT1R (Ad-siAT1R) and a recombinant adenovirus with nonspecific siRNA (Ad-siControl) were constructed to infect the isolated islets for 72 h. AT1R, GLUT-2, and GCK expressions in the islets were investigated and islet perifusion was performed to evaluate the kinetics of insulin release. Results The expression level of AT1R in the isolated islets from db/db mice was twice that of islets from db/m mice. The islets treated with Ad-siAT1R showed significantly decreased AT1R mRNA and protein levels and significantly increased expression of GLUT-2 (by 190%) and GCK (by 121%) compared to those treated with Ad-siControl (P<0.05). In response to stimulation with 16.7 mmol/L glucose, the first-phase insulin secretion was impaired in both Ad-siControl group and mock infected group with the peak insulin levels only 1.8 times of the basal level; the first-phase insulin secretion was markedly improved in islets treated with Ad-siAT1R, with a peak insulin level reaching 2.8 times of the basal level. Conclusions In isolated islets of db/db mice, selective AT1R inhibition can restore the first phase insulin secretion by up-regulating GLUT-2 and GCK, which may be one of the potential mechanisms by which AT1R blockers improve insulin secretion function.