TCY-NH2对脑死亡大鼠胰腺炎性反应和细胞凋亡的影响

Effects of TCY-NH2 on pancreatic inflammatory response and apoptosis in brain dead rats

目的 TCY-NH2对胰腺炎症反应及细胞凋亡的作用尚不明确。文中旨在通过构建大鼠脑死亡模型,探讨TCY-NH2对脑死亡大鼠供胰的保护作用及相关机制。方法 将18只雄性SD大鼠随机分组3组:假手术组,除不加压诱导脑死亡,余操作同脑死亡组;脑死亡组,采用颅内缓慢间断加压诱导大鼠脑死亡,维持脑死亡状态6 h;TCY-NH2组,在加压诱因脑死亡时TCY-NH2(0.6 mg/kg, iv)处理大鼠,余操作同脑死亡组。在脑死亡6 h后留取胰腺标本。免疫组化检测MPO表达。荧光实时定量PCR检测IL-1β、TNF-α、VCAM-1、ICAM-1的mRNA表达。Western blot检测p-IκBα、IκBα、p-p65、Bax、Bcl2和cl-Caspase3蛋白表达。TUNEL法检测细胞凋亡。结果 与脑死亡组胰腺组织浸润的中性粒细胞(4.20±2.79)比较,TCY-NH2组、假手术组(0.70±1.00、0.40±0.66)明显减少(P<0.05);与脑死亡组比较,TCY-NH2组、假手术组的胰腺组织中IL-1β、TNF-α、VCAM-1、ICAM-1的mRNA表达明显降低(P<0.05)。与脑死亡组比较,TCY-NH2组、假手术组的胰腺组织中p-IκBα和p-p65的蛋白表达明显降低(P<0.05),IκBα蛋白表达明显升高(P<0.05)。与脑死亡组胰腺组织中细胞凋亡(2.50±1.96)比较,TCY-NH2组、假手术组(0.60±0.66、0.30±0.46)明显减少(P<0.05)。与脑死亡组比较,TCY-NH2组、假手术组的胰腺组织中Bax和cl-Caspase3的蛋白表达明显降低(P<0.05),Bcl2蛋白表达明显升高(P<0.05)。结论 TCY-NH2可以减轻脑死亡供体胰腺中炎性反应和细胞凋亡,为提高脑死亡供胰质量提供潜在的干预方法。

Objective The effect of TCY-NH2 on pancreatic inflammatory response and apoptosis is still unclear. The objective of this study was to investigate the protective effect of TCY-NH2 on pancreas of brain dead rats and its related mechanism.Methods Eighteen male SD rats were randomly divided into 3 groups: sham operation group, brain death group, and TCY-NH2 group. Rats in sham operation group were operated with the same operation except for brain death induced by no pressure. The brain death of rats in brain death group was induced by slow and intermittent intracranial pressure and maintained for 6 h. Rats in TCY-NH2 groupwere treated with TCY-NH2(0.6 mg/kg, iv) at pressurized induced brain death, and the other treatments were the same as the brain death group. Pancreatic specimens were collected6 h after brain deathrats. MPO expression was detected by immunohistochemistry. The mRNA expression of IL-1β,TNF-α,VCAM-1, and ICAM-1 was detected by real-time PCR. The protein expression of p-IκBα, IκBα, p-p65, Bax, Bcl2, and cl-Caspase3 was detected by the Western blot. Apoptosis was detected by the TUNEL assay. Results Compared with the brain death group(4.20±2.79), the amount of neutrophils infiltrated into pancreatic tissue was significantly reduced in the TCY-NH2 group and the sham operation group(0.70±1.00, 0.40±0.66)(P<0.05);Compared with the brain death group, the mRNA expression of IL-1β, TNF-α, VCAM-1, and ICAM-1 was significantly decreased in the TCY-NH2 group and the sham operation group(P<0.05);Compared with the brain death group, the protein expression of p-IκBα and p-p65 in the TCY-NH2 group was significantly decreased(P<0.05), and the protein expression of IκBα was significantly increased(P<0.05);Compared with the brain death group(2.50±1.96), cell apoptosis in the TCY-NH2 group and the sham operation group(0.60±0.66,0.30±0.46) was significantly reduced(P<0.05);Compared with the brain death group, the protein expression of Bax and cl-Caspase3 in the TCY-NH2 group and the sham operation group was significantly reduced(P<0.05), and the expression of Bcl2 protein was significantly increased(P<0.05). Conclusion TCY-NH2 can reduce the inflammatory response and apoptosis of pancreas in brain dead donors, providing a potential intervention method to improve the quality of brain-dead donor pancreas.