钙超载在大鼠血脂异常胰腺炎中的研究

Research on mechanism of calcium overload in hyperlipidemia pancreatitis model

[目的]探讨钙超载在血脂异常胰腺炎(HLP)发病机制中的作用,从而为维拉帕米治疗HLP提供理论依据。[方法]将80只SD大鼠随机分成4组,每组20只:即假手术(SO)组,重症急性胰腺炎(SAP)组,HLP组,维拉帕米(CCB)治疗组。24h后统计各组死亡率,随即处死大鼠,检测血三酰甘油(TG);采用ELISA检测胰腺组织三磷酸肌醇(IP3);采用流式细胞仪技术测定胰腺细胞内钙离子(Ca2+)水平;采用Western blot检测胰腺组织核转因子(NF)-κB p65蛋白表达;采用ELISA法检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6;苏木精-伊红染色对大鼠胰腺组织学损伤进行评估。[结果]1HLP组、CCB组TG水平均较SO组和SAP组显著升高,差异有统计学意义(P<0.05);2SO组死亡率明显低于其他各组(P<0.05),HLP组与SAP组、CCB组差异有统计学意义(P<0.05);3SO组与SAP组、SAP组与HLP组、HLP组与CCB组胰腺组织内IP3比较均差异有统计学意义(P<0.05);4 SAP组胰腺腺泡细胞Ca2+水平显著高于SO组(P<0.05),HLP组Ca2+高于SAP组、CCB组(P<0.05);5SAP组胰腺组织NF-κBp65表达较SO组显著升高(P<0.05),HLP组较SAP组、CCB组显著升高(P<0.05);6SO组血清TNF-α、IL-6浓度与SAP组相比差异有统计学意义(P<0.05),HLP组较SAP组、CCB组明显升高(P<0.05);7SAP组胰腺组织学评分较SO组,HLP组较SAP组、CCB组均明显升高(P<0.05)。[结论]钙超载在HLP的发生发展中起着关键性作用,维拉帕米治疗HLP效果显著。

[Objective]To explore the role of calcium overload in the pathogeny of hyperlipidemic pancreatitis and provide theoretical foundation for verapamil as a new drug for hyperlipidemic pancreatitis. [Methods]Eighty healthy SD male rats were randomly divided into four groups：Sham operation group（SO group）, Sever acute pancreatitis group （SAP）, Hyperlipidemic pancreatitis group （HLP） and Verapamil Treatment Group（CCB）. The mortality rate of rats in four groups were recorded 24 hours after the model were induced. Then the rats were all sacrificed all and the pancreas tissue and serum were collected. The serum triglyceride was measured. IP3, IL-6 and TNF-α contents in pancrease tissue were detected with ELISA. Intracellular Calcium level in pancrease tissue was measured by flow cytometry. NF-κB expression in pancrease tissue was measured by Western blot. The were detected with the ELISA. Pancreatitis grade was pathologically evaluated by HE stainning. [Results]①Mortality rate of among the groups： there was significant differences between SO group and the three groups（P〈0.05）. The similar difference was noted between HLP group and SAP/CCB group（P〈0.05）. ②Blood TG：TG in HLP and CCB group were signif- icantly increased than SAP and SO group（P〈0.05）.③IP3 expression of pancreatic tissue： HLP group was significantly increased than SAP group and CCB group（P〈0.05）. SO group was significantly lower than SAP group（P〈0.05）. ④ Pancreatic intracellular Calcium level： HLP group was significantly higher than SAP group and CCB group（P〈0.05）. SO group was significantly lower than SAP group（P〈0.05）. ⑤Pancreatic NF-κB content：NF-κB in SAP group was significantly higher than SO group（P〈0.05）. SAP group and CCB group was significantly lower than HLP group（P〈0.05）. ⑥The IL-6 and TNF-α contents：SAP group were significantly increased than SO group（P〈0.05）,SAP group and CCB group was significantly lower than HLP group（P〈0.05）. ⑦Pancreatic pathologic grade：SO group was significantly lower than SAP group（P〈0.05） ,both SAP and CCB group were significantly lower than HLP group（P〈 0.05）. [Conclusion]Calcium overload play a very important role in the development of HLP. Verapamil shows benefit effect in the treatment of HLP.