海马胆碱能神经刺激肽通过活化3型毒蕈碱受体促进INS-1细胞胰岛素释放  

Insulinotropic action of hippocampal cholinergic neurostimulating peptide mediated by activated type 3 muscarinic receptor in INS-1 cells

作　　者：高飞[1] 陈宏[1] 张桦[1] 郭南京[1] 徐艳花[1] 蔡德鸿[1] 

机构地区：[1]南方医科大学珠江医院内分泌科,广东广州510282

出　　处：《南方医科大学学报》2012年第4期580-582,共3页Journal of Southern Medical University

基　　金：中华国际医学交流基金(2100325)

摘　　要：目的明确海马胆碱能神经刺激肽(HCNP)对INS-1细胞胰岛素分泌影响,并对其可能作用的毒蕈碱样胆碱能受体途径进行分析。方法 INS-1细胞随机分为3组:对照组、HCNP组、HCNP+佛波酯(PMA)组。对照组给予RPMI 1640液培养,HCNP组给予人工合成HCNP(50 pg/ml)与RPMI 1640液共培养,HCNP+PMA组予人工合成HCNP与RPMI 1640液共培养后,行胰岛素释放前18 h加入PMA(10 nmol/L);应用放射免疫方法检测各组不同浓度葡萄糖培养液中胰岛素含量。实时定量PCR检测对照组、HCNP组HCNP前体蛋白(HCNP-pp)、胆碱乙酰转移酶(choline acetyltransferase,ChAT)、3型毒蕈碱样受体(3-typemuscarinic receptor,M3R)基因水平表达情况。结果浓度为3.3 mmol/L葡萄糖,三组间胰岛素含量无明显差异。浓度为5.6mmol/L、16.7 mmol/L葡萄糖作用下,HCNP组胰岛素含量均高于对照、HCNP+PMA组,差异具有统计学意义。HCNP组INS-1细胞HCNP-pp、ChAT、M3R三者mRNA含量均高于对照组,差异具有统计学意义。结论 HCNP通过提高ChAT活性,推测影响乙酰胆碱合成,进一步活化M3R,促进胰岛素分泌,PMA可阻断此作用。Objective To characterize the insulinotropic action of hippocampal cholinergic neurostimulating peptide（HCNP） and analyze the role of type 3 muscarinic receptor（M3R） pathway in the action of HCNP.Methods INS-1 cells were incubated in routine RPMI 1640 medium（control group）,RPMI 1640 supplemented with 50 pg/ml synthetic HCNP（HCNP group）,or HCNP-containing medium with the addition of PMA 18 h prior to insulin release assay.The insulin levels in the medium was measured using radioimmunoassay following stimulation with different concentrations of glucose.Real-time quantitative PCR was used for detecting the gene expression of HCNP-pp,choline acetyltransferase（ChAT） and M3R in HCNP group and control group.Results After stimulation with different concentrations of glucose（5.6 and 16.7 mmol/L）,HCNP group showed significantly higher insulin levels than the control and HCNP＋ PMA groups.Compared with those in the control group,the mRNA levels of HCNP-pp,ChAT,and M3R were all lowered in HCNP group.Conclusions HCNP can promote insulin release in INS-1 cells by increasing ChAT activity and activating M3R,and this effect is inhibited by PMA.

关 键 词：海马胆碱能神经刺激肽 INS-1细胞 胰岛素 3型毒蕈碱受体 

分 类 号：R587.1[医药卫生—内分泌]