摘要
目的 基于"肺肠同治"理论研究诃子对肠缺血再灌注损伤(ⅡRI)小鼠肠、肺损伤的治疗作用,并初步探讨诃子发挥此作用是否基于减轻ⅡRI小鼠全身性炎症反应.方法 选取30只8周龄健康雄性C57BL/6J小鼠随机分为5组:假手术(Sham)组,模型(ⅡRI)组,ⅡRI+诃子低、中、高剂量组,每组6只小鼠.采取夹闭肠系膜上动脉45 min、再灌注90 min的方法建立ⅡRI模型.诃子给药各组灌胃不同剂量诃子,1次/d,连续7 d;Sham组开腹不造模,仅在同时间点灌胃生理盐水;ⅡRI组造模后灌胃生理盐水.HE染色法观察小鼠肠、肺组织病理形态变化,TUNEL染色法检测肠道细胞的凋亡情况,ELISA试剂盒检测血清中的TNF-α、IL-1β和IL-6水平.结果 与Sham组相比,ⅡRI组小鼠肠道整体出现水肿、血瘀及黏连现象,肠横切面显示固有层结构破坏,肠黏膜绒毛顶端上皮下间隙明显增宽,绒毛顶端部分脱落,而经诃子给药治疗后整体肠道损伤程度得到显著改善;ⅡRI组小鼠Chiu's评分升高(P＜0.01),而诃子给药组Chiu's评分显著降低(P＜0.01);ⅡRI组小鼠肠道细胞凋亡数量明显增多,而诃子给药组小鼠肠道细胞凋亡数量减少;ⅡRI组小鼠肺组织切片HE结果出现明显的肺泡壁增厚和大量炎性细胞浸润肺泡间隙的情况,而诃子给药组的肺损伤情况得到明显改善.与Sham组相比,ⅡRI组小鼠肺损伤评分显著增高(P＜0.01),而诃子给药后肺损伤评分降低,差异具有统计学意义(P＜0.05,P＜0.01);ⅡRI组小鼠肺湿/干质量比比值增高(P＜0.05),而诃子给药组小鼠肺湿/干质量比比值降低(P＜0.05),肺水肿程度得到显著改善;ⅡRI组小鼠血清中促炎因子TNF-α、IL-1β和IL-6水平显著升高(P＜0.01),而诃子剂量依赖性地降低小鼠血清中TNF-α、IL-1β和IL-6水平,且差异具有统计学意义(P＜0.01).结论 诃子可有效减轻小鼠ⅡRI,抑制肠道细胞凋亡,同时能减轻小鼠肠缺血再灌注后引起的肺部损伤,发挥肺肠同治作用,该作用与诃子降低血清中TNF-α、IL-1β和IL-6水平,减轻小鼠体内炎症反应有关.
Abstract
Objective To study the therapeutic effect of T.chebula on intestinal ischemia-reperfusion injury(ⅡRI)in mice based on the theory of"simultaneous treatment of lung and intestine",and to preliminarily discuss whether the effect of T.chebule is based on its reduction of systemic inflammatory response in ⅡRI mice.Methods Thirty healthy male C57BL/6J mice aged 8 weeks were randomly divided into 5 groups:Sham group,model(ⅡRI)group,ⅡRI+T.chebula low,medium and high dose groups,with 6 mice in each group.The ⅡRI model was established by clamping the superior mesenteric artery for 45 min and reperfusion for 90 min.After establishing the ⅡRI model,each group was administrated with different doses of T.chebula once a day for 7 d.Sham group was given normal saline by gavage at the same time point after laparotomy without modeling.ⅡRI group was given normal saline by intragastric administration after modeling.The pathological and morphological changes of intestinal and lung tissues were observed by HE staining,the apoptosis of intestinal cells was detected by TUNEL staining,and the levels of TNF-α,IL-1β and IL-6 in serum were detected by ELISA kit.Results Compared with the Sham group,the intestinal tract of mice in the ⅡRI group showed edema,blood stasis and adhesion as a whole,the intestinal transverse section showed structural damage of lamina propria,the subepithelial space of the intestinal mucosal villi tip was significantly widened,and the villi tip was partially shed,while the overall intestinal injury degree was significantly improved after treatment with T.chebula.Compared with the Sham group,Chiu's score was increased in the ⅡRI group(P＜0.01),while Chiu's score was significantly decreased in T.chebula-treated group(P＜0.01).Compared with the Sham group,the number of intestinal cell apoptosis in the ⅡRI group was significantly increased,while the number of intestinal cell apoptosis in T.chebula-treated group was reduced.Compared with the Sham group,HE results of the ⅡRI group showed obvious thickening of alveolar wall and infiltration of a large number of inflammatory cells in the alveolar space,while the lung injury was significantly improved in T.chebula-treated group.Compared with the Sham group,the lung injury score of mice in the ⅡRI group was significantly increased(P＜0.01),while the lung injury score was decreased after administration of T.chebula,and the difference was statistically significant(P＜0.05,P＜0.01).Compared with the Sham group,lung wet/dry weight ratio in the ⅡRI group was increased(P＜0.05),while that in T.chebula-treated group was decreased,and the degree of pulmonary edema was significantly improved.Compared with the Sham group,the serum levels of pro-inflammatory factors TNF-α,IL-lβ and IL-6 in the ⅡRI group were significantly increased(P＜0.01),while the serum levels of TNF-α,IL-1β and IL-6 in T.chebula-treated group were decreased in a dose-dependent manner,and the difference was statistically significant(P＜0.01).Conclusion T.chebula can effectively alleviate ⅡRI in mice,inhibit intestinal cell apoptosis,and reduce lung injury caused by intestinal ischemia-reperfusion in mice.It plays the role of simultaneous treatment of lung and intestine,which is related to reducing the serum levels of TNF-α,IL-1β and IL-6 and alleviating inflammatory responses in mice.
基金项目
国家中医药管理局科技专项课题(GZY-KJS-2023-026)
陕西省中医药管理局"秦药"研发重点实验室项目(2021-QYPT-003)
陕西省中医药管理局中医药"双链融合"中青年创新团队建设项目(2022-SLRH-YQ-004)
陕西省科技厅现代新药中药研发项目(2022YWZX-PG-01)
空军军医大学伙伴实验室合作交流计划项目(2023HB018)
国家科技期刊平台
NSTL
万方数据