清下化瘀方改善重症急性胰腺炎大鼠肠黏膜屏障功能对p38MAPK/ATF2信号通路的影响

【作者】 孔婧； 王晓素； 周秉舵； 李英； 徐亭亭；

【机构】 上海中医药大学附属岳阳中西医结合医院； 上海中医药大学脾胃病研究所；

【摘要】 目的:通过动物实验探讨清下化瘀方通过改善肠屏障功能治疗急性胰腺炎的分子生物学机制,以阐明其在中西医结合疗法治疗本病中的作用基础。方法:采用逆胆胰管注射牛磺胆酸钠的方法建立重症急性胰腺炎大鼠模型,分别予清下化瘀方及善宁干预后取材,观察及检测大鼠肠黏膜屏障通透性改变(肠黏膜上皮损伤程度),并分析p38MAPK/ATF2表达的变化,明确清下化瘀方作用机理。结果:(1)造模及胰腺组织病理损伤;造模24h后,模型组大鼠胰腺病理总积分模型组明显高于空白组及假手术组(P<0.01),出现广泛凝固性坏死,间质水肿明显,大量单核和中性粒细胞浸润,提示造模成功。(2)肠上皮损伤指数比较:模型组与其余四组相比(P<0.01),提示模型组肠黏膜损伤明显增加,而中药组及善宁组对肠黏膜损伤有改善作用。中药组与善宁组两者疗效相当(P=>0.05)。(3)肠屏障功能变化:常规透射电镜下观察显示,空白组及假手术组组肠黏膜上皮细胞表面微绒毛排列整齐,实验各组回肠出现不同程度的线粒体肿胀,嵴不清,内质网肿胀,上皮细胞微绒毛脱落,上皮细胞连接增宽。(4)p38MAPK/ATF2表达变化:免疫组化测定各组大鼠胰腺p38MPK、ATF2表达方面,造模24h后大鼠胰腺组织出现p38MAPK阳性表达,模型组表达水平明显较高(P<0.01),同时善宁组阳性表达与空白组及假手术组比较有统计学意义(P<0.01),但中药及善宁组表达较模型组显著降低(P<0.01)。造模后ATF2表达也里上升趋势,模型组较其它组表达明显(P<0.01)。结论:实验表明清下化瘀方能够减轻重症急性胰腺炎大鼠的肠黏膜屏障的损伤,从而减少肠源性内毒素血症的产生,可通过调控p38MAPK/ATF2表达,减轻机体炎症反应,从而达到减少胰腺损伤,治疗SAP的目的。更多还原

【Abstract】 Objective:To explore the molecular biology mechanism of Qing-xia-hua-yu Decoction to treat acute pancreatitis through the improvement of intestinal barrier function by animal experiment,which act to clarify in the foundation about the treatment in traditional Chinese medicine and Western medicine combination therapy.Method:The SAP model of rat was induced by retrograde injection of sodium taurocholate into the biliopancreatic duct,which were treated with Qing-xia-hua-yu Decoction and Sandostatin intervention materials,observation and detection of rat intestinal mucosa barrier permeability change(intestinal mucosal injury degree),and analyze the changes p38 MAPK/ATF2 signaling pathway at the molecular level and tissue morphology angle,to identify Qing-xia-hua-yu Decoction mechanism.Results:(1) SAP model and pancreatic tissue pathological injury:24 h after modeling,total scores of pancreatic pathology in model group was significantly higher than that of control group and sham operation group(P<0.01),suggesting that the model was made successfully.(2) intestinal epithelial damage index:that the model group,compared to the rest of the four groups(P<0.01),the intestinal mucosal injury increased significantly,and traditional Chinese medicine group and SS group on intestinal mucosa injury improved.The effect of traditional Chinese medicine group and Sandostatin group are the same(P=>0.05),no statistically significant differences.(3)Changes of intestinal barrier function:routine transmission electron microscope observation showed that,blank group and sham operation group of intestinal mucosal epithelial cell surface microvilli arranged in neat rows,the ileum in three experimental groups appeared different degree of swelling of mitochondria and crest is not clear.(4) P38 MAPK/ATF2 changes:by immunohistochemical method for the determination of rats pancreatic p38 MAPK and Atf2 expression,positive expression of p38 MAPK in pancreatic tissue of rats in model group.The expression levels were significantly higher(P<0.01),also positive expression of Sandostatin group have statistical significance compared with control group and sham operation group(P<0.01),but the Chinese medicine group and Sandostatin group expression was much lower than that of model group decreased significantly(P<0.01).After modeling,the expression of ATF2 also showed an upward trend,the model group was higher than that of other groups(P<0.01).Conclusions:The animal experiment showed that Qing-xia-hua-yu Decoction can alleviate the damage of intestinal mucosa barrier in rats with severe acute pancreatitis,thus reduce the production of intestinal endotoxemia,can reduce the inflammatory reaction by regulating p38 MAPK/ATF2 signaling pathway,so as to decrease the damage to the pancreas,in the treatment of SAP.更多还原