基于IL-9/IL-9R途径研究针灸保护DSS诱导的UC大鼠肠黏膜屏障的作用机制  

Study of the mechanism of acupuncture and moxibustion in protecting the intestinal mucosal barrier in DSS-induced UC rats based on the IL-9/IL-9R pathway

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作  者:黄艳 孙可鑫 徐静 马喆 赵继梦 李灵杰 赵琛 杨玲 李峰 刘雅楠 张玥 吴璐一 黄儒德 HUANG Yan;SUN Kexin;XU Jing;MA Zhe;ZHAO Jimeng;LI Lingjie;ZHAO Chen;YANG Ling;LI Feng;LIU Yanan;ZHANG Yue;WU Luyi;HUANG Rude(Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China)

机构地区:[1]Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China [2]Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China

出  处:《Journal of Acupuncture and Tuina Science》2024年第2期91-103,共13页针灸推拿医学(英文版)

基  金:国家自然科学基金项目,No.82174501,No.82105012,No.82004475,No.81873372;上海市自然科学基金项目,No.21ZR1460200,No.22ZR1458400;上海市人才发展基金项目,No.2021058;上海中医药大学“杏林百人计划”项目,No.[2020]23。

摘  要:目的:观察针灸对溃疡性结肠炎(UC)大鼠结肠组织肠黏膜白细胞介素(IL)-9/IL-9受体(IL-9R)的调节作用,探讨针灸对UC大鼠肠黏膜屏障的保护机制。方法:将雄性Sprague-Dawley大鼠随机分为正常组和造模组。造模组先予以4%葡聚糖硫酸钠(DSS)水溶液连续自由饮用7 d。大鼠UC模型构建成功后,将造模组大鼠随机分为模型组、隔药灸组和电针组。隔药灸组和电针组分别接受隔药灸和电针双侧天枢穴干预,每日1次,连续干预7 d。采用苏木素-伊红(HE)染色法观察大鼠结肠组织病理学变化并评分;采用酶联免疫吸附测定法测定血清中IL-9、IL-6、IL-1β和血红蛋白H(HbH)的蛋白含量;采用免疫印迹法或免疫组化法检测大鼠结肠组织IL-9、IL-9R、claudin-2、紧密连接蛋白1(ZO-1)和occludin的蛋白表达;采用免疫荧光法检测PU.1、CD4与IL-9蛋白共表达。结果:与正常组比较,模型组大鼠结肠组织损伤严重,出现大面积的溃疡,伴有充血水肿,结肠组织病理学评分显著升高(P<0.01),血清HbH浓度降低(P<0.01),血清中IL-9、IL-6和IL-1β蛋白含量均升高(P<0.01),结肠组织中ZO-1和occludin蛋白表达降低(P<0.01),结肠组织中IL-9、IL-9R蛋白表达均增高(P<0.05),结肠组织中IL-9/PU.1和IL-9/CD4阳性共表达增加(P<0.05)。与模型组相比,隔药灸组和电针组大鼠结肠黏膜结构逐渐修复,可见愈合性溃疡,结肠组织病理学评分降低(P<0.05),血清HbH浓度增加(P<0.01),血清IL-9、IL-6和IL-1β蛋白含量均降低(P<0.05),结肠组织中ZO-1和occludin蛋白表达升高(P<0.05),结肠组织中IL-9、IL-9R蛋白表达降低(P<0.01),结肠组织中IL-9/PU.1和IL-9/CD4阳性共表达降低(P<0.05)。结论:隔药灸和电针均能通过调节转录因子PU.1抑制UC结肠组织中IL-9与IL-9R的蛋白表达,促进肠黏膜屏障修复,降低血清中促炎因子IL-9、IL-6和IL-1β的蛋白含量,这可能是针灸减轻UC结肠黏膜炎症,保护肠黏膜屏障的关键机制之Objective:To observe the effects of acupuncture and moxibustion on interleukin(IL)-9/IL-9 receptor(IL-9R)in the colon tissue of rats with ulcerative colitis(UC)and investigate the protective mechanism of acupuncture and moxibustion on the intestinal mucosal barrier in UC rats.Methods:Male Sprague-Dawley rats were randomly divided into a normal control(NC)group and a modeling group.UC models were prepared by giving 4%dextran sulfate sodium(DSS)water for 7 d.After the successful construction of the UC rat model,the modeling group was randomly divided into a UC group,a herb-insulated moxibustion(HM)group,and an electroacupuncture(EA)group.HM and EA interventions at bilateral Tianshu(ST25)were performed once a day for 7 d.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in the colon.The serum concentrations of IL-9,IL-6,IL-1β,and hemoglobin-H(HbH)were determined by enzyme-linked immunosorbent assay.The protein expression levels of IL-9,IL-9R,claudin-2,zonula occludens-1(ZO-1),and occludin in the colon tissue were measured by Western blotting or immunohistochemistry.Immunofluorescence was used to detect the co-expression of PU.1 and CD4 with the IL-9 protein.Results:Compared with the NC group,the colon tissue of UC rats was severely damaged and ulcerated with congestion and edema,and the colonic histopathological score increased significantly(P<0.01).The serum HbH concentration decreased significantly(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1βincreased(P<0.01).The protein expression of colonic ZO-1 and occludin decreased significantly(P<0.01),while the protein expression of colonic IL-9 and IL-9R increased(P<0.05).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 increased in the colon tissue(P<0.05).Compared with the UC group,the colonic mucosal structures were gradually repaired in both HM group and EA group,and healed ulcers could be observed,the colonic histopathological score decreased significantly(P<0.05).The serum concentration of HbH increased(P<0.

关 键 词:针刺疗法 电针 药饼灸疗法  天枢 结肠炎 溃疡性 肠黏膜屏障 白细胞介素9 大鼠 

分 类 号:R2-03[医药卫生—中医学]

 

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