大麻素2型受体激动剂JWH133调节小鼠胰腺腺泡细胞钙信号及治疗急性胰腺炎的机制  

Mechanisms of Cannabinoid 2 receptor agonist JWH133-mediated modulations in cell calcium signals of mouse pancreatic acinar cells and its application for treatment of acute pancreatitis

作　　者：夏坤锟[1,2,3] 郭文治 吴杰 张水军 王贵宪 袁维堂[1] Xia Kunkun;Guo Wenzhi;Wu Jie;Zhang Shuijun;Wang Guixian;Yuan Weitang(Department of Colon and Rectal Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Henan Key Laboratory of Digestive Organ Transplantation,Zhengzhou 450052,China;Open and Key Laboratory of Hepatobiliary & Pancreatic Surgery and Digestive Organ Transplantation at Henan Universities,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,郑州450052 [3]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450052

出　　处：《中华实验外科杂志》2019年第5期861-863,共3页Chinese Journal of Experimental Surgery

基　　金：河南省高等学校重点科研项目计划(19A320043);河南省重点研发与推广专项科技攻关项目(192102310411).

摘　　要：目的观察JWH133对乙酰胆碱(ACh)引起的Ca^2+振荡的影响,探讨其作用机制及靶点。方法酶解法分离小鼠胰腺腺泡细胞,利用膜片钳全细胞记录,通过细胞外给药方式,在分离的胰腺腺泡细胞上观察10μmol/L JWH133对10 nmol/L ACh引起Ca^2+振荡的影响,检测JWH133的药理学作用。记录10μmol/L JWH133对10 mmol/L左旋精氨酸(L-Arg)引起的Ca^2+振荡增强作用的影响。通过L-Arg诱发小鼠急性胰腺炎模型,观察并测量胰腺组织结构,细胞形态及出血坏死情况。结果JWH133作用的靶点为细胞表面的大麻素2型受体(CB2R)。10μmol/L的JWH133能将10 nmol/L ACh引起Ca^2+振荡的标准化净电流减少到(26.0±4.2)%(P<0.01,n=9)。在CB2R敲除(CB2R-KO)小鼠,与给药前比较,施加10μmol/L的JWH133后,ACh引起Ca^2+振荡的净电流(标准化后)为(97.0±6.7)%(P>0.05,n=9)。应用10μmol/L的JWH133则可将10 mmol/L L-Arg引起增强的Ca^2+振荡的标准化静电流减少至(106.0±16.7)%(P<0.01,n=10)。此外,病理切片显示JWH133预处理的胰腺组织在水肿、炎症和坏死水平均有所改善,病理学评分差异有统计学意义(P<0.05,n=5)。结论JWH133是通过特异性抑制细胞外的CB2R来抑制细胞内Ca2+信号。JWH133对L-Arg诱发的小鼠急性胰腺炎模型的病理有改善作用。Objective To evaluate the effect of JWH133 on acetylcholine(Ach)-induced Ca^2+oscillations,and to elucidate its mechanism and target.Methods Pancreatic acinar cells were isolated by enzymatic hydrolysis.Patch-clamp whole cell recording techniques were used to measure the effects of 10μmol/L JWH133 on 10 nmol/L ACh-induced intracellular Ca^2+oscillations in pancreatic acinar cells acutely dissociated from wild-type(WT),and Cannabinoid 2 receptor(CB2R)knockout(KO)mice,and the pharmacological effects of JWH133 on the Ca^2+oscillations.The effect of 10μmol/L JWH133 on the enhancement of Ca^2+oscillation induced by 10 mmol/L L-Arginine(L-Arg)was measured.The model of acute pancreatitis in mice was induced by L-Arg,and the histological structure,cell morphology and hemorrhagic necrosis were examined.Results The results showed that JWH133 could inhibit ACh-induced Ca^2+oscillations likely through CB2R.Patch-clamp recordings demonstrated that 10μmol/L of JWH133 could reduce the 10 nmol/L ACh-induced Ca2+oscillations current to(26.0±4.2)%(P<0.01,n=9).In CB2R KO mice,the inhibitory effects of JWH133 could reduce the 30 nmol/L ACh-induced Ca^2+oscillations current to(97.0±6.7)%(P>0.05,n=9).10μmol/L of JWH133 attenuated the enhancement of calcium oscillations current to(106.0±16.7)%(P<0.01,n=10)induced by 10 mmol/L L-Arg.In L-Arg acute pancreatitis model,JWH133 improved L-Arg-induced pancreatic pathological changes(P<0.05,n=5).Conclusion JWH133 inhibits intracellular calcium signals by selectively inhibiting extracellular CB2R.In L-Arg-induced acute mouse pancreatitis model,JWH133 significantly improves pathological changes.All these results suggest that JWH133 is a promising candidate for treatment of acute pancreatitis.

关 键 词：急性胰腺炎 大麻素2型受体激动剂 钙信号 膜片钳 

分 类 号：R576[医药卫生—消化系统]