粉防己碱对脂多糖致大鼠胰腺腺泡细胞损伤的保护作用及其机制研究  

Study on protective effect of tetrandrine on lipopolysaccharide induced pancreatic acinar cell damage and its mechanism

作　　者：张红[1] 李永渝[2] 吴咸中[3] 

机构地区：[1]陕西中医学院,咸阳712046 [2]同济大学医学院,上海200092 [3]天津医科大学,300070

出　　处：《中国危重病急救医学》2006年第3期157-160,共4页Chinese Critical Care Medicine

基　　金：国家自然科学基金资助项目(30370643)

摘　　要：目的探讨粉防己碱(T et)对脂多糖(LPS)诱发大鼠胰腺腺泡细胞损伤的保护作用及其机制。方法胶原酶法分离雄性SD大鼠胰腺腺泡细胞,预先经T et(50μm o l/L、100μm o l/L)处理15 m in后,再经LPS(10 m g/L)或正常培养液处理,在0、1、4和10 h采集上清液,检测其中丙二醛(M DA)含量及超氧化物歧化酶(SOD)、磷脂酶A2(PLA2)活性;采用四甲基偶氮唑蓝比色法(M TT)检测胰腺腺泡细胞存活情况;部分胰腺腺泡细胞经F luo 3/AM荧光探针负载后,于相应时间点采用灌流方式给予T et或LPS,激光共聚焦显微镜观察单个胰腺腺泡细胞内钙离子浓度(〔C a2+〕i)。结果T et可减轻LPS所致的细胞损伤(P均<0.05),抑制LPS诱发的胰腺腺泡细胞〔C a2+〕i升高(P均<0.05),降低细胞培养上清液中M DA含量和PLA2活性、增加SOD活性。结论T et可能通过抑制钙超载、增强抗氧化能力以及减少胰酶活化,减少LPS所致胰腺腺泡细胞损伤,从而发挥对胰腺腺泡细胞的保护作用。Objective To evaluate the protective effect of tetrandrine （Tet） on lipopolysaccharide （LPS） -induced pancreatic acinar cell damage and to explore its mechanism. Methods SD male rat＇s pancreatic acinar cells were isolated by collagenase digestion and they were pre - treated with Tet （50μmol/L and 100μmol/L）, then exposed to LPS （10mg/L） or conventional culture medium respectively. At 0, 1, 4, 10 hours after treatment with the agents, cell viability was determined by methyl thiazolyl tetrazolium （MTT）, and the supernatant supernate of cells was collected for determination of the content of malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） and phospholipase A2 （PLA2）. Some cells were loaded with Fluo- 3/AM, and the dynamic change in intracellular Ca^2＋ （C[a^2＋3]i） in single pancreatic acinar cell was determined by laser scanning confocal microscopy. Results Tet attenuated LPS -induced cell damage （P〈0. 05 and P〈0. 01） and inhibited the elevation of cytosolic free calcium of rat pancreatic acinar cells. In the supernatant, Tet pretreatment decreased the content of MDA and the activity of PLA2 and increased the activity of SOD （all P〈0. 05）. Conclusion Tet attenuates LPS - induced cell damage by blocking [Ca^2＋]i overload, inhibiting superoxidative response,decreasing activity of pancreatic enzyme, thus it shows a protective effect on pancreatic acinar cells.

关 键 词：粉防己碱 脂多糖 胰腺 腺泡细胞 钙 

分 类 号：R285[医药卫生—中药学]