维拉帕米治疗高脂血症胰腺炎机制研究

Research on mechanism of verapamil in treating hyperlipidemic pancreatitis

目的探讨维拉帕米治疗高脂血症胰腺炎(HLP)的机制。方法将80只SD大鼠随机分为假手术组(SO组)、重症急性胰腺炎模型组(SAP组)、HLP模型组(HLP组)、维拉帕米治疗组(CCB组)四组,每组20只。24 h后统计大鼠死亡率,随即处死大鼠,取出胰腺组织和穿刺心脏收集血液,检测三酰甘油(TG)水平。采用ELISA法检测胰腺组织三磷酸肌醇(IP3)、血清血小板活化因子(PFA)和血栓素A2(TXA2)水平,流式细胞术测定胰腺细胞内钙离子水平,酸滴定法检测血清磷脂酶A2(PLA2),HE染色评估大鼠胰腺组织学损伤情况。结果 (1)24 h死亡率:SO组24 h死亡率低于其他三组,差异均有统计学意义(P<0.05);HLP组24 h死亡率高于SAP、CCB组,差异均有统计学意义(P<0.05)。(2)TG水平:HLP组、CCB组TG水平均较SO组及SAP组显著升高,差异均有统计学意义(P<0.05)。(3)胰腺组织IP3水平:SO组与SAP组、SAP组与HLP组、HLP组与CCB组胰腺组织内IP3水平比较,差异均有统计学意义(P<0.05)。(4)胰腺腺泡细胞钙离子水平:SAP组钙离子水平高于SO组,HLP组钙离子水平高于SAP组,而CCB组钙离子水平低于HLP组,差异均有统计学意义(P<0.05)。(5)血清PLA2水平:SAP组血清PLA2水平较SO组显著升高,而HLP组PLA2水平较SAP组、CCB组显著升高,差异均有统计学意义(P<0.05)。(6)血清PAF、TXA2水平:SO组与SAP组血清PAF、TXA2水平比较,差异均有统计学意义(P<0.05);HLP组血清PAF、TXA2水平较SAP组和CCB组明显升高,差异均有统计学意义(P<0.05)。(7)胰腺病理学评分:SAP组胰腺损伤评分较SO组显著升高,差异有统计学意义(P<0.05);HLP组胰腺损伤评分较SAP组和CCB组明显升高,差异均有统计学意义(P<0.05)。结论钙超载在大鼠HLP的发生、发展中具有重要作用。维拉帕米治疗大鼠HLP效果显著。

Objective To explore the mechanism of verapamil intreating hyperlipidemic pancreatitis（HLP）. Methods Eighty SD rats were randomly divided into the sham operation group（SO）,severe acute pancreatitis group（SAP）,HLP model group and verapamil treatment group（CCB）,20 cases in each group. The mortality rate was calculated after 24 h, then rats were killed for taking pancreatic tissue and collecting blood by puncture. The blood triglyceride（TG） level was detected. The levels of IP3 in pancreatic tissue,serum PFA and TXA2 were detected by ELISA. The flow cytometry was used to measure pancreatic intracellular calcium ion level. The acid titration method was used to measure serum PLA2. The pancreatic histological injury was evaluated by HE staining. Results（1）The 24 h mortality rate：the mortality rate in the SO group was lower than that in the other three groups with statistical difference（P〈0.05）. The 24 h mortality rate in the HLP group was higher than that in the SAP and CCB groups with statistical difference（P〈0.05）.（2） The TG level：the TG level in the HLP and CCB groups was significantly increased compared with the SAP and SO groups with statistical difference（P〈0.05）.（3）The IP3 level in pancreatic tissue：the IP3 level in pancreatic tissue had statistical differences between the SO and SAP groups,between the SAP and HLP groups and HLP and CCB groups（P〈0.05）.（4）The calcium ion level in pancreatic acinar cell：the calcium ion level in the SAP group was higher than that in the SO group,the calcium ion level in the HLP group was higher than that in the SAP group,while the calcium ion level in the CCB group was lower than that in the HLP group with statistical difference（P〈0.05）.（5）The serum PLA2 level： the serum PLA2 level in the SAP group was significantly higher than that in the SO group,whereas serum PLA2 level in the HLP group was significantly increased compared with the SAP and CCB groups with statistical difference（P〈0.05）.（6）The serum PAF and TXA2 levels： the serum PAF and TXA2 levels had statistical differences between the SO and SAP groups（P〈0.05）;the serum PAF and TXA2 levels in the HLP group was significantly increased compared with the SAP and CCB groups with statistical difference（P〈0.05）.（7）The pancreatic histological scores： the injury in the SAP group was significantly increased compared with the SO group with statistical difference（P〈0.05）;the pancreatic injuries score in the HLP group was significantly increased compared with the SAP and CCB groups with the statistical differences（P〈0.05）. Conclusion Calcium overloading plays an important role in the development of HLP rat models.Verapamil has remarkable effect in the treatment of HLP rat models.