不同大麻素配体调节小鼠胰腺腺泡细胞中乙酰胆碱诱导的Ca^2＋信号的研究  

Effects of cannabinoid receptor ligands on acetylcholine - induced calcium oscillations in mouse pancreatic acinar cells

作　　者：夏坤锟[1,2] 郭文治 袁维堂 吴杰[2] 张水军 Xia Kunkun;Guo Wenzhi;Yuan Weitang;Wu Jie;Zhang Shuljun(Department of Colon and Rectal Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Chin;Henan Key Laboratory of Digestive Organ Transplantation, Open and Key Laboratory of Hepatobiliary ＆ Pa~reatic Surgery and Digestive Organ Transplantation at Henan Univer- sities, Zhengzhou Key Laboratory of Hepatobiliary ＆ Pancreatic Diseases and Organ Transplantation, Zhengzhou 450052, China)

机构地区：[1]郑州大学第一附属医院结直肠外科,450052 [2]河南省消化器官移植重点实验室河南省高等学校肝胆胰外科与消化器官移植重点学科开放实验室郑州市肝胆胰疾病与器官移植医学重点实验室,郑州450052

出　　处：《中华实验外科杂志》2018年第6期1074-1076,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察各种CBR配体激动剂对急性分离的胰腺腺泡细胞中乙酰胆碱（ACh）诱导的Ca^2＋振荡的影响。方法使用膜片钳全细胞记录，检测大麻素受体1（CB1R）、大麻素受体2（CB2R）和非CBR激动剂对ACh诱发的Ca^2＋振荡的影响。利用野生型（WT）、CB1R敲除（KO）和CB2R-KO小鼠，分别对部分CB2R激动剂和非选择性CBR激动剂（WIN）对小鼠胰腺腺泡细胞Ca^2＋信号的影响进一步检测。结果3种CB2R激动剂均可抑制腺泡细胞内Ca^2＋信号，10 μmol/L的GW，GP1a和JWH133能分别将30 nmol/L ACh引起Ca^2＋振荡的净电流（标准化后）减少至（23.0±4.4）% （P＝0.000，n＝7），（11.0±4.6）% （P＝0.000，n＝6）和（26.0±4.2）% （P＝0.000，n＝9）。而在CB2R-KO小鼠，这三种CB2R激动剂的抑制作用被消除。在WT、CB1R-KO及CB2R-KO小鼠中，非选择性CBR激动剂WIN55，212-2（10 μmol/L）对ACh（30 nmol/L）引起的Ca^2＋振荡的标准化静电流分别减少到（7.0±3.5）% （P＝0.000，n＝6），（5.0±6.9）%（P＝0.000，n＝6）和（5.0±9.3）%（P＝0.000，n＝6）。而内源性大麻样物质2-AG则对Ca^2＋振荡没有影响。结论大麻素受体活性物质调节小鼠的胰腺腺泡细胞中的Ca^2＋信号呈多样性，包括CBR和非CBR，CB2R而不是CB1R在调节小鼠胰腺腺泡细胞中ACh诱导的Ca^2＋振荡起主导作用。Objective To investigate the effects of various CBR ligands on acetylcholine （Ach） -induced Ca^2＋ oscillations in acute isolated pancreatic acinar cells.Methods Patch-clamp whole-cell recordings were applied to measure ACh-induced intracellular Ca^2＋ oscillations in pancreatic acinar cells acutely dissociated from wild-type （WT）, cannabinoid type 1 receptor （CB1R） knockout （KO）, and cannabinoid type 2 receptor （CB2R） KO mice, and the pharmacological effects of various cannabinoid ligands on the Ca^2＋ oscillations were examined.Results Patch-clamp recordings demonstrated that all tested three CB2R agonists inhibited intracellular Ca^2＋ oscillations, 10 μmol/L of GW, GP1a, and JWH133 can reduced the 30 nmol/L ACh-induced Ca^2＋ oscillations current to （23.0±4.4）% （P=0.000, n=7）, （11.0±4.6）% （P=0.000, n=6） and （26.0±4.2）% （P=0.000, n=9） respectively. In CB2R KO mice, the inhibitory effects of these 3 CB2R agonists were abolished, suggesting that their effect is mediated through the CB2R. In WT, CB1R KO and CB2R KO mice, Non-selective CB receptor agonists WIN55, 212-2 also inhibited Ca^2＋ oscillations, 10 μmol/L of WIN55, 212-2 can reduced the 30 nmol/L ACh-induced Ca^2＋ oscillations current to （7.0±3.5）% （P=0.000, n=6）, （5.0±6.9）% （P=0.000, n=6） and （5.0±9.3）% （P=0.000, n=6） respectively. but endogenous Non-selective cannabinoid substance, 2-AG did not show the inhibitory effect.Conclusion Cannabinoid ligands modulate mouse pancreatic acinar cell Ca^2＋ oscillations in a complex manner, through CB receptors or non-CB receptors. CB2R rather than CB1R plays a critical role in the regulation of Ca^2＋ signals in mouse pancreatic acinar cells.

关 键 词：大麻素受体 钙信号 胰腺腺泡细胞 膜片钳 

分 类 号：R285.5[医药卫生—中药学]