柴芩承气汤对急性胰腺炎大鼠胰腺腺泡细胞的保护作用及其机制

Protective effects of Chaiqin Chengqi Decoction on isolated pancreatic acinar cells in acute pancreatitis rats and the mechanisms

目的:探讨柴芩承气汤(Chaiqing Chengqi Decoction,CQCQD)对急性胰腺炎(acute pancreatitis,AP)大鼠胰腺腺泡细胞的保护作用及其机制。方法:SD大鼠以CQCQD灌喂,制备柴芩承气汤含药血清(CQCQserum,CQCQS);SD大鼠分为AP组和假手术组,分离胰腺腺泡细胞并与CQCQS共同孵育,采用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法检测不同浓度CQCQS对腺泡细胞存活力的影响,并用钙荧光指示剂Fluo-3-AM加载腺泡细胞,激光共聚焦显微镜直接观测腺泡细胞内钙荧光强度(fluorescent intensity,FI),定量分析FI并以此表示[Ca2+]i。结果:5%和10%CQCQS均可提高AP大鼠胰腺腺泡细胞存活力(P<0.05),且10%CQCQS提高细胞存活力的作用比5%CQCQS更为显著(P<0.05);[Ca2+]i随AP病程延长而升高(P<0.05),CQCQS可抑制AP大鼠胰腺腺泡细胞内[Ca2+]i升高(P<0.05)。结论:CQCQD对AP时的胰腺细胞有保护作用,其机制可能与抑制胰腺腺泡细胞内钙超载有关。

Objective： To investigate the protective effects of Chaiqin Chengqi Decoction （CQCQD）, a compound traditional Chinese herbal medicine, on acinar cells and the changes of intracellular calcium concentration （[Ca^2＋]i） in acute pancreatitis （AP） rats. Methods： CQCQ serum （CQCQS） was prepared by intragastric administration of CQCQD in SD rats, and AP was induced in SD rats by biliary-pancreatic duct ligation method. Pancreatic acinar cells were isolated by collagenase digestion and co-incubated with the drug serum containing CQCQD in vitro, then cell viability was determined by methyl thiazolyl tetrazolium （MTT） method and intracellular fluorescent intensity （FI） was observed and analyzed to investigate the changes of intracellular calcium concentration （[Ca^2＋]i）. Results： Both 5% and 10% CQCQS could improve the viability of pancreatic acinar cells in AP rats （P〈0.05）, and 10% CQCQS was more effective than 5 % CQCQS （P〈0.05）. [Ca^2＋]i was elevated inAP rats （ P〈0.05）, while the level of [Ca^2＋]i elevation was reduced after CQCQS treatment （P〈0. 05）. Conclusion： CQCQD has a protective effect on pancreatic acinar cells of AP rats, which may be associated with its inhibition effect on intracellular calcium overload.