白三烯B4及瞬时电位香草酸亚型受体1变化在急性胰腺炎中的作用及大黄干预研究

Experiment on the changes of leukotriene B4 and TRPV1 in acute pancreatitis and the intervention of rhubarb

[目的]观察急性胰腺炎大鼠LTB4、TRPV1、CGRP等相关指标变化,探讨大黄防治急性胰腺炎的可能机理。[方法]清洁级SD大鼠80只,按体重随机分为4组,正常组、模型组、TRPV1拮抗剂组、大黄治疗组。采用胆胰管开口十二指肠结扎法制备重症胰腺炎模型,术后3h开始分组给药,术后12h处死全部大鼠,腹主动脉取血并收集胰腺组织备测。应用光镜检测胰腺组织超微结构变化,应用ELISA技术检测LTB4变化,应用Real-time PCR技术检测TRPV1、CGRP变化。[结果]正常组胰腺肉眼见胰腺呈浅粉红色;模型组大鼠肉眼见胰腺实质大片坏死、出血和脂肪变性,部分可见含血腹水,光镜下见胰腺腺体结构紊乱,小叶间隔明显增大,腺泡可见大片凝固性坏死并伴有出血,坏死灶及周围有大量炎细胞浸润;TRPV1拮抗剂组大鼠胰腺组织出血及水肿较模型组减轻,少量腹水,浑浊程度低,光镜下胰腺实质可见少许坏死、出血及少量炎性细胞浸润。大黄治疗组未见腹水,病理变化轻,皂化亦明显减轻,胰腺水肿、出血,坏死减轻,其组织病理学评分较模型组下降,有统计学意义(P<0.05)。模型组见血淀粉酶、LTB4表达明显上升,分别为5386.72±190.34、476.52±21.33;TRPV1拮抗剂组血淀粉酶、LTB4表达下降,分别为2315.67±135.42、263.68±15.69,较模型组有统计学意义。大黄治疗组血淀粉酶、LTB4表达明显下降,分别为2136.82±126.72、237.45±12.56,较模型组有统计学意义(P<0.05)。模型组胰腺组织TRPV1、CGRP mRNA表达上升,分别为3.59±1.23、3.08±1.53;TRPV1拮抗剂组TRPV1、CGRP mRNA表达下降,分别为1.82±0.34、1.57±0.26;与模型组比较有统计学意义(P<0.05)。大黄治疗组TRPV1、CGRP mRNA表达明显下降,分别为1.51±0.23、1.28±0.15,较模型组有统计学意义(P<0.05)。[结论]大黄可能通过调节LTB4、TRPV1等炎症因子的表达能发挥防治急性胰腺炎的效用。

[Objective]To investigate the mechanism of acute pancreatitis by observing the changes of indexes related to LTB4-TRPV1 signaling pathway based on the treatment of rhubarb.[Methods]80clean SD rats were randomly divided into normal group,blank group,TRPV1 antagonist group,rhubarb treatment group depending on body mass.Except normal group,other rats were administrated with different treatments at the time of 3hafter modeling surgery,and then blood from abdominal aorta was taken and pancreas tissue was collected at the time of 12 h.The changes of pancreatic tissue histopathology were observed with light microscope,the changes of LTB4 were assayed by ELISA,and the expression of TRPV1,CGRP was detected by Real time PCR.[Results]The pancreas was light pink in normal group by naked eyes,while in the blank group it appeared necrosis,hemorrhage,and steatosis.Moreover,part of model rats were found ascites containing with blood.There were pancreatic gland disorders,increased interlobular septa,gland parenchyma necrosis and hemorrhage,inflammatory cell infiltrating in pancreatic tissue under light microscope.In the TRPV1 antagonist group,the pancreatic tissue hyperemia and edema was relieved compared with the blank group,and the ascites was also less and cleaner.When observed under light microscope the pancreatic parenchyma appeared little necrosis,hemorrhage and inflammatory cell infiltrating.The Rhubarb treatment group rats appeared no ascites,little saponification and slight edema,hemorrhage and necrosis in pancreatic tissue under light microscope.The histopathology score was decreased than those in the blank group（P〈0.05）.The expression of blood amylase,LTB4,and TRPV1 and CGRP mRNA in the blank group was increased,which obviously decreased in TRPV1 antagonist group and rhubarb treatment group（P〈0.05）.[Conclusion]Rhubarb can regulate the expression of LTB4 and TRPV1to treat the acute pancreatitis.