维拉帕米减轻脂多糖导致的大鼠胰腺腺泡细胞损伤机制的研究

目的：研究维拉帕米（Vera）对脂多糖（LPS）导致的大鼠胰腺腺泡细胞损伤的拮抗作用及其机制。 方法：胶原酶法分离大鼠胰腺腺泡细胞，预先经Vera (50 mg/L、100 mg/L) 处理15min后, 再经LPS (10 mg/L)或正常培养液处理，在不同的时点（30 min、1 h、4 h、10 h）采集上清液,检测其中丙二醛含量、超氧化物歧化酶、磷脂酶A2的活性；采用MTT 法检测胰腺腺泡细胞的活性；部分胰腺腺泡细胞经Fluo-3/AM负载后，于相应的时点采用灌流方式给予药物或刺激剂，激光共聚焦显微镜观察单个胰腺腺泡细胞［Ca2+］i的变化。 结果: Vera可减轻LPS所致的细胞损伤（P<0.05）；抑制LPS诱发的胰腺腺泡细胞 ［Ca2+］i 升高（P<0.05）；降低细胞培养上清液中丙二醛含量和磷脂酶A2的活性、增强超氧化物歧化酶的活性。 结论：Vera可能通过抑制钙超载、增强抗氧化能力以及减少胰酶活化的机制，减少LPS所致的胰腺腺泡细胞损伤，从而发挥对胰腺腺泡细胞的保护作用。

Mechanisms by which verapamil protects against lipopolysaccharide-induced pancreatic acinar cell damage in vitro

AIM: To investigate the protective effect of verapamil (Vera) on lipopolysaccharide (LPS)-induced pancreatic acini damage and its mechanism. METHODS: Rat pancreatic acinar cells were isolated by collagenase digestion and were pre-treated with Vera (50 mg/L, 100 mg/L), then exposed to LPS (10 mg/L) or normal culture media, respectively. At various time points (30 min, 1 h, 2 h, 4 h, 10 h) after treatment of the agents, cell viability was determined by MTT and supernatant of cell culture was collected to measurer the content of maloidialdehyde (MDA), the activity of superoxide dismutase (SOD) and phospholipase (PLA2). Some cells were loaded with Fluo-3/AM, and the dynamic change of Ca2 ]i in single pancreatic acinar cell was determined by laser scanning confocal microscopy. RESULTS: Vera attenuated LPS-induced cell damage (P<0.05) and inhibited the elevation of cytosolic free calcium in rat pancreatic acinar cells. In the supernatant, Vera decreased the content of MDA and the activity of PLA2 (P<0.05) and increased the activity of SOD (P<0.05). CONCLUSION: Vera attenuates LPS-induced cell damage by blocking calcium overload, inhibiting superoxidative response, decreasing activity of pancreatic enzyme, and hence plays a protective effect on pancreatic acinar cells.