CAJ | 학술논문

目的观察双孔钾通道KCNK16在大鼠胰岛中的表达情况,并探讨促进β细胞胰岛素分泌的刺激因素对其表达的调节.方法采用实时定量PCR和组织化学染色技术观察KCNK16在大鼠胰岛及其他组织中的表达.分别用葡萄糖、催乳素、软脂酸以及曲古菌素A(TrichostatinA,TSA)处理大鼠胰岛24h,实时定量PCR和Western印迹法检测KCNK16的表达水平.以ELISA检测TSA处理大鼠胰岛24 h后葡萄糖和KC1刺激的胰岛素分泌水平.不同浓度和时间的TSA处理INS1-E细胞,实时定量PCR检测KCNK16的表达水平.结果 KCNK16在大鼠胰岛中高表达.葡萄糖、催乳素和脂肪酸并不影响大鼠胰岛KCNK16的表达.TSA显著增强大鼠胰岛葡萄糖和KC1刺激的胰岛素分泌(P＜0.05),同时显著下调KCNK16的mRNA和蛋白表达(P＜0.01).在INS1-E细胞,TSA抑制KCNK16的表达作用呈浓度和时间依赖性(P＜0.05).结论 KCNK16在大鼠胰岛中特异性高表达,且其表达水平能被TSA显著下调,KCNK16表达的下调有可能介导了TSA增强的胰岛素分泌.
목적 관찰쌍공갑통도KCNK16재대서이도중적표체정황,병탐토촉진β세포이도소분비적자격인소대기표체적조절.방법 채용실시정량PCR화조직화학염색기술관찰KCNK16재대서이도급기타조직중적표체.분별용포도당、최유소、연지산이급곡고균소A(TrichostatinA,TSA)처리대서이도24h,실시정량PCR화Western인적법검측KCNK16적표체수평.이ELISA검측TSA처리대서이도24 h후포도당화KC1자격적이도소분비수평.불동농도화시간적TSA처리INS1-E세포,실시정량PCR검측KCNK16적표체수평.결과 KCNK16재대서이도중고표체.포도당、최유소화지방산병불영향대서이도KCNK16적표체.TSA현저증강대서이도포도당화KC1자격적이도소분비(P＜0.05),동시현저하조KCNK16적mRNA화단백표체(P＜0.01).재INS1-E세포,TSA억제KCNK16적표체작용정농도화시간의뢰성(P＜0.05).결론 KCNK16재대서이도중특이성고표체,차기표체수평능피TSA현저하조,KCNK16표체적하조유가능개도료TSA증강적이도소분비.
Objective To observe the expression of a two-pore domain K+ channel KCNK16 in rat islets,and to explore its regulation by insulin secretagogues.Methods Real time-PCR and immunohistochemistry were employed to observe the expression of KCNK16 in rat islets and other tissues.Rat islets were exposed to glucose,prolactin,palmitate,and trichostatin A (TSA) for 24 h and KCNK16 expression were determined by realtime-PCR and Western blot.Glucose-and KCl-stimulated insulin secretion were assayed by ELISA after islets were exposed to TSA for 24 h.The dose-and time-dependent effects of TSA on KCNK16 expression in INS1-E cells were determined by RTPCR.Results KCNK16 was highly expressed in rat islets.Glucose,prolactin,and palmitate did not affect the expression of KCNK16 in rat islets.TSA treatment significantly potentiated glucose-and KCl-stimulated insulin secretion in isolated rat islets (P ＜ 0.05),along with decreased KCNK16 mRNA and protein expressions (P ＜ 0.01).TSA inhibited KCNK16 expression in a dose-and time-dependent manner in INS1-E cells (P＜0.05).Conclusion KCNK16 is exclusively and highly expressed in rat islets and its expression can be down-regulated by TSA treatment.It is possible that TSA enhances insulin secretion via decreasing KCNK16 expression.