胰泰复方对慢性胰腺炎胰腺纤维化大鼠腺泡细胞凋亡的内质网应激状态的影响

Effects of Yitai Compound(胰泰复方)on Endoplasmic Reticulum Stress in Rats with Chronic Pancreatitis and Pancreatic Fibrosis

目的观察胰泰复方对慢性胰腺炎(chronic pancreatitis,CP)胰腺纤维化大鼠腺泡细胞凋亡的内质网应激状态的影响。方法将60只SD(Sprague Dawley,SD)大鼠随机分为模型组,清胰利胆颗粒组,胰泰复方低、中、高剂量组与空白组。空白组大鼠腹腔注射生理盐水,其余各组腹腔注射13.3%L-精氨酸,造模后灌胃给药,各组连续干预30 d取血清及胰腺组织,并观察血清淀粉酶的变化,苏木精-伊红染色(hematoxylin-eosin staining)和Masson(马松三色)染色观察组织病理形态学改变,免疫组化法及Western blot法检测胰腺腺泡细胞中环磷酸腺苷反应元件结合转录因子同源蛋白(C/EBP homology protein,CHOP)及葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)含量。结果免疫组化检测结果表明,CHOP及GRP78表达空白组与模型组比较有统计学意义(P<0.05);胰中组、对照组与模型组比较差异有统计学意义(P<0.05)。各治疗组相较于模型组血清淀粉酶差异有统计学意义(P<0.05),Western Blot检测结果显示,模型组CHOP及GRP78蛋白表达显著上升,治疗后,胰高组、胰中组、对照组CHOP及GRP78蛋白表达显著减少,与模型组比较有统计学意义(P<0.05)。结论胰泰复方可以通过下调内质网应激状态,进而减少胰腺腺泡细胞的凋亡,从而干预CP胰腺纤维化的进展。

Objective To observe the effect of Yitai Compound(胰泰复方)on endoplasmic reticulum stress state of acinar cell apoptosis in chronic pancreatitis rats with pancreatic fibrosis.Methods Sixty Sprague Dawley rats were randomly divided into model group,Qingyi Lidan Granule(清胰利胆颗粒)group,Yitai Compound low,medium and high dose groups and blank group.Rats in the blank group were intraperitoneally injected with normal saline,and the other groups were intraperitoneally injected with 13.3%L-arginine.After modeling,the rats were given intraperitoneal administration.Serum and pancreatic tissue were collected in each group for 30 days of continuous intervention,and the changes of serum amylase were observed.Hematoxylin eosin staining and Masson staining were used to observe the histopathological changes.Immunohistochemical and Western blot methods were used to detect the contents of cyclic adenosine monophosphate reaction element binding transcription factor(CHOP)and glucose regulated protein 78(GRP78)in pancreatic acinar cells.Results The results of immunohistochemistry showed that the expressions of CHOP and GRP78 in the blank group and the model group were statistically significant(P<0.05).There were significant differences among the Yitai Compound middle dose group,the control group and the model group(P<0.05).Compared with the model group,each treatment group had a significant difference in serum amylase(P<0.05).The results of Western blot showed that the protein expressions of CHOP and GRP78 in the model group were significantly increased.The expres-sions of CHOP and GRP78 in the model group were increased significantly.After treatment,the expressions of CHOP and GRP78 in the Yitai Compound high and middle dose groups and control group were decreased significantly,which was statistically significant compared with those of the model group(P<0.05).Conclusion Yitai Compound can reduce the apoptosis of pancreatic acinar cells by down-regulating the stress state of endoplasmic reticulum,thereby intervening the progression of pancreatic fibrosis of chronic pancreatitis.