基于以方测证理论法探讨脾胃虚寒型十二指肠溃疡实验模型的构建及模型评价研究
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摘要
目的:基于以方测证理论构建可靠、稳定的十二指肠溃疡(DU)脾胃虚寒证候大鼠实验模型,并对模型进行综合评价。方法:将48只SD大鼠随机分为正常组、模型组、柴胡疏肝散6.56g/kg组(非方证对应组)和黄芪建中汤9.27g/kg组(方证对应组)。采用苦寒泻下加劳倦过度法构建脾胃虚寒证候,运用阿司匹林加无水乙醇建立DU模型。观测大鼠的整体状态、体质量、进食量和肛温变化,计算溃疡指数(UI),HE染色观察十二指肠黏膜病理改变,ELISA法检测血清PGE、NO、MDA含量,免疫组化法测定TLR-2、MyD88蛋白表达。结果:与正常组比较,模型组大鼠体质量、进食量、肛温均明显降低。给予黄芪建中汤治疗后与模型组比较,大鼠整体状态恢复正常,体质量增加、进食量增多、肛温升高。与正常组比较,模型大鼠UI明显增加,小肠绒毛受损严重;给予黄芪建中汤治疗后,大鼠UI较模型组显著降低,黏膜形态恢复正常。与正常组比较,模型大鼠血清NO、PGE含量显著降低,MDA含量明显增加,TLR-2及MyD88蛋白表达显著上调;给予黄芪建中汤治疗后与模型组比较,大鼠血清NO、PGE含量明显增加,MDA含量显著降低,TLR-2及MyD88蛋白表达明显下调。结论:本研究构建了一种稳定、可重现的脾胃虚寒型DU病证结合大鼠实验模型,该模型可为十二指肠溃疡疾病的实验研究提供新的载体。
Objective: To establish a reliable and stable experimental model of duodenal ulcer rat with Piweixuhan syndrome and evaluate the model. Methods: 48 SD rats were randomly divided into the normal group,the model group,6. 56 g/kg Chaihu Shugan San group,and 9. 27 g/kg Huangqi Jianzhong Decoction( HQJZ) group. Piweixuhan syndrome model was established by using methods of " bitter cold induced diarrhea" and " overstrain". Duodenal ulcer model was established by using absolute ethyl alcohol and aspirin. The general symptoms and signs of rats were observed and the changes of body weight,food intake and rectal temperature were monitored. The index of duodenal ulcer( UI) was evaluated. The pathological changes of duodenal mucosa were observed by HE staining. The levels of NO,PGE and MDA in serum were detected by ELISA,and the expressions of TLR-2 and MyD88 were detected by immunohistochemistry. Results: Compared with the normal group,the body weight,food intake,and rectal temperature in all other groups were decreased. After HQJZ treatment,the body weight,food intake and rectal temperature were increased. Compared with the normal group,the ulcer index( UI) in the model group significantly increased,and HE staining showed that the intestinal villi were seriously damaged. After the treatment with HQJZ,UI of rats was significantly decreased,and the morphology of duodenal mucosa recovered. Compared with the normal group,the contents of NO and PGE in serum of model group was significantly decreased,the content of MDA and the expressions of TLR-2 and MyD88 protein were significantly increased.After the treatment with HQJZ,the levels of NO and PGE in serum were significantly increased,the level of MDA was significantly decreased,and the expressions of TLR-2 and MyD88 protein were significantly decreased. Conclusion: The results show that the duodenal ulcer rat model with Piweixuhan syndrome was successfully established,which may provide a new method for the experimental study of duodenal ulcer disease.
Objective: To establish a reliable and stable experimental model of duodenal ulcer rat with Piweixuhan syndrome and evaluate the model. Methods: 48 SD rats were randomly divided into the normal group,the model group,6. 56 g/kg Chaihu Shugan San group,and 9. 27 g/kg Huangqi Jianzhong Decoction( HQJZ) group. Piweixuhan syndrome model was established by using methods of " bitter cold induced diarrhea" and " overstrain". Duodenal ulcer model was established by using absolute ethyl alcohol and aspirin. The general symptoms and signs of rats were observed and the changes of body weight,food intake and rectal temperature were monitored. The index of duodenal ulcer( UI) was evaluated. The pathological changes of duodenal mucosa were observed by HE staining. The levels of NO,PGE and MDA in serum were detected by ELISA,and the expressions of TLR-2 and MyD88 were detected by immunohistochemistry. Results: Compared with the normal group,the body weight,food intake,and rectal temperature in all other groups were decreased. After HQJZ treatment,the body weight,food intake and rectal temperature were increased. Compared with the normal group,the ulcer index( UI) in the model group significantly increased,and HE staining showed that the intestinal villi were seriously damaged. After the treatment with HQJZ,UI of rats was significantly decreased,and the morphology of duodenal mucosa recovered. Compared with the normal group,the contents of NO and PGE in serum of model group was significantly decreased,the content of MDA and the expressions of TLR-2 and MyD88 protein were significantly increased.After the treatment with HQJZ,the levels of NO and PGE in serum were significantly increased,the level of MDA was significantly decreased,and the expressions of TLR-2 and MyD88 protein were significantly decreased. Conclusion: The results show that the duodenal ulcer rat model with Piweixuhan syndrome was successfully established,which may provide a new method for the experimental study of duodenal ulcer disease.
引文
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[11]Afroz S, Yagi A, Fujikawa K, et al. Lysophosphatidic acid in medicinal herbs enhances prostaglandin E2 and protects against indomethacin-induced gastric cell damage in vivo and in vitro . Prostaglandins Other Lipid Mediat, 2018, 135(1)∶36～44.
[12]Gong J, Zhang Z, Zhang X, et al. Effects and possible mechanisms of Alpinia officinarum ethanol extract on indomethacin-induced gastric injury in rats . Pharm Biol, 2018, 56(1)∶294～301.
[13]Liu W, Shang P, Liu T, et al. Gastroprotective effects of chebulagic acid against ethanol-induced gastric injury in rats . Chem Biol Interact, 2017, 278(1)∶1～8.
[14]Hackam D J, Sodhi C P. Toll-Like Receptor-Mediated Intestinal Inflammatory Imbalance in the Pathogenesis of Necrotizing Enterocolitis . Cell Mol Gastroenterol Hepatol, 2018, 6(2)∶229～238.
[15]李芹, 张会永, 吴天石, 等. 脾阳虚证动物模型造模方法与模型评价的研究进展 . 世界科学技术-中医药现代化, 2015, 17(8)∶1721～1728.
[16]Saghaei F, Karimi I, Jouyban A, et al. Effects of captopril on the cysteamine-induced duodenal ulcer in the rat. Exp Toxicol Pathol, 2012, 64(4)∶373～377.
[2]Budimir I, Stojsavljevic S, Hrabar D, et al. Bleeding Peptic Ulcer - Tertiary Center Experience: Epidemiology, Treatment and Prognosis . Acta Clin Croat, 2017, 56(4)∶707～714.
[3]Roberts-Thomson IC. Rise and fall of peptic ulceration: A disease of civilization? . J Gastroenterol Hepatol, 2018, 33(7)∶1321～1326.
[4]吴刚, 郭胜红, 袁华兵, 等. 黄芪建中汤加味方联合西药治疗消化性溃疡脾胃虚寒证疗效观察 . 现代中西医结合杂志, 2018, 27(11)∶1203～1205.
[5]魏晏, 魏明. 黄芪建中汤联合督脉灸治疗消化性溃疡临床研究 . 中医学报, 2015, 30(205)∶874～877.
[6]林路平, 邝卫红. 许鑫梅教授治疗消化性溃疡复发经验 . 广州中医药大学学报, 2013, 30(1)∶105～111.
[7]孙静晶, 赵晓丹, 廉艳红, 等. "阳道实, 阴道虚"理论与消化性溃疡 . 环球中医药, 2015, 8(2)∶194～196.
[8]张声生, 王垂杰, 李玉锋, 等. 消化性溃疡中医诊疗专家共识意见(2017年) . 中华中医药杂志, 2017, 32(9)∶4089～4093.
[9]李军祥, 陈誩, 肖冰, 等. 消化性溃疡中西医结合诊疗共识意见(2017年) . 中国中西医结合消化杂志, 2018, 26(2)∶112～120.
[10]郑筱萸. 中药新药临床研究指导原则. 北京: 中国医药科技出版社, 2002∶151～155.
[11]Afroz S, Yagi A, Fujikawa K, et al. Lysophosphatidic acid in medicinal herbs enhances prostaglandin E2 and protects against indomethacin-induced gastric cell damage in vivo and in vitro . Prostaglandins Other Lipid Mediat, 2018, 135(1)∶36～44.
[12]Gong J, Zhang Z, Zhang X, et al. Effects and possible mechanisms of Alpinia officinarum ethanol extract on indomethacin-induced gastric injury in rats . Pharm Biol, 2018, 56(1)∶294～301.
[13]Liu W, Shang P, Liu T, et al. Gastroprotective effects of chebulagic acid against ethanol-induced gastric injury in rats . Chem Biol Interact, 2017, 278(1)∶1～8.
[14]Hackam D J, Sodhi C P. Toll-Like Receptor-Mediated Intestinal Inflammatory Imbalance in the Pathogenesis of Necrotizing Enterocolitis . Cell Mol Gastroenterol Hepatol, 2018, 6(2)∶229～238.
[15]李芹, 张会永, 吴天石, 等. 脾阳虚证动物模型造模方法与模型评价的研究进展 . 世界科学技术-中医药现代化, 2015, 17(8)∶1721～1728.
[16]Saghaei F, Karimi I, Jouyban A, et al. Effects of captopril on the cysteamine-induced duodenal ulcer in the rat. Exp Toxicol Pathol, 2012, 64(4)∶373～377.