芸香苷静脉输注对大鼠急性胰腺炎腺泡细胞凋亡的影响及机制

目的探讨芸香苷(Rutoside,Ru)静脉输注给药对实验性急性胰腺炎(acute pancreatitis,AP)腺泡细胞凋亡的影响及作用机制。方法3%牛磺胆酸钠(sodium taurocholate,STC)逆行胆胰管注射诱导大鼠AP模型,取胰腺做病理学检查。应用末端脱氧核苷酸转移酶介导的末端标记(terminadeoxynucleotidyl transferase biotin-dUTP nick end labeling,TUNEL)方法检测胰腺组织中的腺泡细胞凋亡。用免疫组化法检测Fas、FasL蛋白的表达。结果Ru15、30、60mg·kg-1·h-13个静脉输注剂量组可改善胰腺的病理组织学变化,且凋亡指数(apoptosis index,AI)高于模型组〔6h Ru大、中、小剂量治疗组分别为(23·33%±3·49%)、(21·56%±3·81%)、(16·24%±2·83%),6h模型组(8·54%±1·28%),P<0·01〕;Fas蛋白阳性表达细胞的灰度值低于模型组〔6h Ru大、中剂量治疗组分别为(189·17±5·79)、(203·83±10·94),6h模型组(226·86±10·37),P<0·01〕;FasL在各组腺泡细胞中均有不同程度的表达,但各治疗组的阳性表达细胞的灰度值高于模型组〔6h Ru大、中、小剂量治疗组分别为(220·63±7·41)、(218·72±4·48)、(207·22±22·67),6h模型组(181·70±9·01),P<0·05,P<0·01〕,各组的AI在各时间点均与Fas阳性表达细胞的灰度值呈正相关,与FasL阳性表达细胞的灰度值和胰腺严重程度呈负相关(P<0·05,P<0·01)。结论Ru对实验性AP的保护作用可能与促腺泡细胞凋亡有关,Fas/FasL系统可能参与此过程而诱导细胞凋亡。

Effect of Rutoside on pancreatic acinar cell apoptosis in rats with acute pancreatitis and its mechanism

Aim To study the effect of Rutoside (Ru) on pancreatic acinar cell apoptosis in rats with acute pancreatitis(AP). Methods The AP model in rats was induced by retrograde injection of 3% sodium taurocholate into biliopancreatic duct. Ru (15, 30 and 60 mg·kg~ -1 ·h~ -1 )was administered by intravenous infusion for 6 hours immediately after the induction of AP.The histopathological changes of pancreas were observed under light microscope and electronic microscope.A terminal deoxynucleotidyl transferase-mediated dUTP-biotion nick end labeling(TUNEL) method was used to detect apotosis of pancreatic acinar cell and apoptosis immunohistochemical method. Results Ru (15, 30 and 60 mg·kg~ -1 ·h~ -1 ) improved the histopathological changes of pancreas significantly.The apotosis index of pancreatic acinar cells was significantly higher than that the AP model group. The gray value of Fas was lower than that of the AP model group. Immuno-staining revealed that moderate to strong FasL immunoreactivity was present in the cytoplasm of the acini of all groups.But in AP model group,the gray value of FasL was lower than that in Ru-treated groups.AI was positively correlated with the gray value of Fas,and was negatively correlated with the gray value of FasL and the severity of pancreas damages. Conclusion The protective effect of Ru on AP may be concerned with the induction of apoptosis in injured pancreatic acinar cells.And the Fas/FasL system may contribute to the process.