白芍总苷对重症急性胰腺炎大鼠胰腺病理学改变及NF-κB通路的影响

Effect of total paeony glycoside on pathological changes of pancreas and NF-κB pathway in rats with severe acute pancreatitis

目的研究白芍总苷(TGP)对重症急性胰腺炎(SAP)大鼠胰腺病理学改变的影响及其作用机制。方法180只SD大鼠随机分为假手术组、SAP组、TGP(20、40、60 mg/kg)组和加贝酯(gabexate mesylate,GM)10 mg/kg组,每组30只;采用逆行胰胆管注射浓度为3.5%牛磺胆酸钠(1 mL/kg)的方法制备SAP大鼠模型,造模后立即尾静脉注射药物。24 h后,观察各组大鼠胰腺组织大体外观变化,计算胰腺湿/干比重,生化分析法检测血清淀粉酶(AMS)和脂肪酶(LPS)水平;通过HE染色法观察胰腺组织病理学改变并进行病变评分,TUENL法观察胰腺细胞凋亡状况并计算凋亡指数(apoptosis index,AI);ELISA法测定胰腺组织炎症细胞因子(TNF-α、IL-1β、IL-6)水平;Western blot法检测核因子-κB(NF-κB)、激活型半胱胺酸蛋白酶-3(cleaved caspase-3)、c-Jun氨基端激酶(p-JNK)、B淋巴细胞瘤-2(bcl-2)、bcl-2相关X蛋白(Bax)表达。结果与SAP组比较,经TGP 40、60 mg/kg或GM 10 mg/kg干预明显改善SAP大鼠胰腺水肿、充血、皂化斑等变化;降低胰腺湿/干比重(P<0.01);降低血清AMS、LPS水平(P<0.05);显著改善腺泡水肿破裂、炎性细胞浸润、细胞坏死等胰腺组织病理学改变,降低病变评分(P<0.01);显著改善胰腺细胞凋亡状况,降低AI(P<0.01);降低胰腺组织TNF-α、IL-1β、IL-6水平(P<0.05);显著下调NF-κB、bcl-2表达而上调Cleaved Caspase-3、p-JNK表达(P<0.05)。与GM 10 mg/kg组比较,TGP 60 mg/kg组对SAP大鼠胰腺大体外观、组织病理学改变的改善作用较优,胰腺湿/干比重、AMS和LPS含量、病变评分、凋亡指数、TNF-α和IL-1β水平降低(P<0.05),NF-κB表达下调而Cleaved Caspase-3、p-JNK表达上调(P<0.05)。结论TGP具有改善SAP大鼠胰腺病理学改变的作用,其机制可能与抑制NF-κB信号通路而抑制炎症反应、促进胰腺细胞凋亡有关。

Objective To investigate the effect of total glucosides of paeony(TGP)on pathological changes of pancreas in rats with severe acute pancreatitis(SAP)and its mechanism.Methods A total of 180 SD rats were randomly divided into sham operation group,SAP group,TGP(20,40,60 mg/kg)groups and gabexate mesilate(GM)10 mg/kg group,30 in each group.The SAP rat model was SAP rat model was established by retrograde pancreaticobiliary duct injection of 3.5%taurocholic acid(1ml/kg),and the drugs were given by tail vein injection immediately after the model was completed.After 24 hours,the changes of the appearance of pancreas tissue were observed,the wet/dry specific gravity of pancreas was calculated,the content of AMS and LPS in serum were detected by biochemical analysis.The pathological change of pancreas tissue was examined by HE staining and lesions was scored;the apoptosis was observed by TUNEL method and the apooptosis index(AI)was calculated.The contents of TNF-α,IL-1β,IL-6 were detected by ELISA.The expression of NF-κB,cleaved caspase-3,p-JNK,bcl-2,Bax were determined by Western blot.Results Compared with SAP group,TGP 40,60 mg/kg or GM 10 mg/kg the changes of pancreatic edema,hyperemia and saponification spots were significantly improved,and the wet/dry specific gravity of pancreas was decreased(P<0.01).The serum levels of AMS and LPS were decreased(P<0.05).The pathological changes such as acinar edema,rupture,inflammatory cell infiltration and cell necrosis were significantly improved,and the lesion score was decreased(P<0.01).The apoptosis of pancreatic cells was significantly improved and AI was decreased(P<0.01).The levels of TNF-α,IL-1βand IL-6 in pancreatic tissue were significantly decreased(P<0.05).The expression of NF-κB and Bcl-2 was significantly down-regulated,while the expression of cleaved caspase-3 and p-JNK was up-regulated(P<0.05).Compared with GM 10 mg/kg group,TGP 60 mg/kg group had better effect on improving the appearance and histopathology of pancreas in SAP rats.The wet/dry ratio,AMS and LPS content,lesion score,AI,TNF-αlevel and IL-1βlevel were decreased(P<0.05).The expression of NF-κB was down-regulated and the expression of cleaved caspase-3,p-JNK were up-regulated(P<0.05).Conclusion TGP can improve the pathological changes of the pancreas in SAP rats,and its mechanism may be related to inhibiting NF-κB signaling pathway,inhibiting inflammatory reaction and promoting apoptosis of pancreatic cells.