艾灸对克罗恩病大鼠结肠肿瘤坏死因子-α含量及艾灸鼠结肠上清液对培养的结肠上皮细胞内紧密连接蛋白及其基因表达的影响

目的:通过观察温和灸、隔药灸对肿瘤坏死因子-α(TNF-α)体外诱导大鼠结肠上皮细胞紧密连接--咬合蛋白(occludin)、闭合蛋白1(claudin-1)和闭锁蛋白1(zonula occludens-1,ZO-1)及其基因表达的影响,探讨艾灸对肠上皮屏障损伤的修复/保护机制.方法:将SD大鼠随机分成正常组、模型组、温和灸组、隔药灸组和西药组,各12只.采用三硝基苯磺酸制备克罗恩病(Crohn's Disease,CD)大鼠模型.温和灸组与隔药灸组选择"天枢""气海"穴分别予以温和灸和隔药灸,每日1次,共治疗14次;西药组以柳氮磺胺吡嘧溶液灌胃,每日2次,共14次.治疗结束后剖取各CD组大鼠结肠,制备结肠黏膜上清液;正常组纯化并培养结肠上皮细胞建立肠上皮屏障体外模型.将各CD组结肠黏膜上清液分别与正常大鼠结肠上皮细胞混合培养1周,采用ELISA法检测各组结肠黏膜上清液TNF-α含量,Western blot和荧光定量PCR法观察其对CD大鼠体外结肠上皮紧密连接occludin、claudin-1和ZO-1及其mRNA表达的影响.结果:CD模型组较正常组大鼠结肠黏膜上清液中TNF-α含量有显著增高(P<0.01),温和灸、隔药灸和柳氮磺胺吡嘧治疗后TNF-α含量显著降低(均P<0.01),其中温和灸、隔药灸组优于西药组(均P<0.05);与此同时,温和灸、隔药灸和西药组结肠黏膜TNF-α上清液体外诱导大鼠结肠上皮紧密连接occludin、claudin-1和ZO-1及其mRNA表达较模型组有显著增高(P<0.01,P<0.05),且隔药灸、温和灸两组也优于西药组(均P<0.05).结论:艾灸(隔药灸、温和灸)可能是通过降低CD大鼠结肠黏膜异常增高的TNF-α,进而促进或调节结肠上皮紧密连接.ccludin,claudin 1和ZO-1及其mRNA表达,达到修复/保护肠上皮屏障损伤的目的.     

隔药灸对肿瘤坏死因子-α介导克罗恩病肠上皮细胞凋亡途径的影响

目的:观察隔药灸对肿瘤坏死因子-α(TNF-α)-肿瘤坏死因子受体相关死亡结构域(TRADD)-Fas相关的死亡结构域(FADD)途径介导克罗恩病(Crohn's disease,CD)肠上皮细胞凋亡的影响,探讨其作用机制。方法:将48只SD雄性大鼠随机分成正常组、模型组、隔药灸组和西药组,每组12只。采用2,4,6-三硝基苯磺酸(TNBS)制备CD模型,造模成功后隔药灸组采用隔药饼灸"天枢""气海"穴,每日1次,每次每穴隔药灸2壮,共灸10d。西药组采用美沙拉嗪灌胃,每日2次,每次2mL,共灌胃10d。治疗结束后取各组大鼠结肠上皮组织,分离、纯化和培养结肠上皮细胞建立体外肠上皮细胞屏障模型,将100ng/mL TNF-α分别与各组肠上皮细胞孵育培养24h,应用荧光黄透过率检测各组肠上皮细胞屏障通透性,Western blot法检测各组肠上皮细胞凋亡途径相关蛋白肿瘤坏死因子受体1(TNFR1)、TRADD、受体作用蛋白1(RIP1)、FADD、锌指蛋白A20(A20)表达以及流式细胞术观察各组肠上皮细胞凋亡情况。结果:与正常组比较,模型组的荧光黄透过率显著升高(P0.001),结肠上皮细胞TNFR1、TRADD、RIP1表达量明显升高(P0.01),A20表达量明显降低(P0.01),细胞凋亡率显著增高(P0.001)。与模型组比较,隔药灸组和西药组的荧光黄透过率显著降低(P0.001),结肠上皮细胞TRADD、RIP1、FADD表达量明显降低(P0.01),A20表达量明显升高(P0.01),结肠上皮细胞凋亡率显著降低(P0.001)。结论:隔药灸可能是通过调控TNF-α介导CD肠上皮细胞凋亡途径的异常,达到保护或修复CD肠上皮屏障损伤之效应。     

隔药灸对克罗恩病大鼠结肠黏膜巨噬细胞M1/M2型极化的影响

目的:观察隔药灸对克罗恩病(CD)大鼠结肠黏膜巨噬细胞M1/M2型极化及炎性因子表达的影响，探讨其作用机制。方法:将40只雄性SD大鼠随机分为正常组、模型组、西药组、隔药灸组，每组10只。采用2,4,6-三硝基苯磺酸灌肠法制备CD模型。造模成功后隔药灸组大鼠给予隔药饼灸“天枢”“气海”,每次每穴灸2壮，每日1次，共10 d。西药组大鼠给予美沙拉嗪灌胃，每日2次，共10 d。治疗结束后取各组大鼠结肠上皮组织，分离、纯化和培养结肠黏膜固有层巨噬细胞。HE染色法观察结肠组织病理变化；透射电镜观察结肠组织超微结构；Western blot法检测结肠黏膜巨噬细胞α7烟碱型乙酰胆碱受体(α7nAChR)、核因子κB(NF-κB) p65、肿瘤坏死因子(TNF)-α的表达水平；流式细胞术检测结肠黏膜组织M1、M2型巨噬细胞的数量；免疫荧光法检测结肠黏膜组织M1、M2型巨噬细胞的表达情况。结果:与正常组比较，模型组大鼠结肠组织呈现巨大溃疡及炎性表现，结肠上皮细胞连接疏松，细胞器结构破坏；结肠黏膜巨噬细胞α7nAChR表达水平降低(P<0.01),NF-κB p65和TNF-α表达水平升高(P&l...     

艾灸对克罗恩病大鼠结肠黏膜MCP-1和IL-8蛋白表达的影响

目的通过观察艾灸对克罗恩病（CD）大鼠结肠黏膜MCP—1和IL-8蛋白表达的影响，探讨艾灸对CD大鼠结肠黏膜免疫功能的作用及可能机制。方法采用国际公认的MorriS方法制备CD大鼠模型。将动物随机分为正常对照组、模型组、隔药灸组、温和灸组、烟条灸组和热水灸组，均选取天枢（双侧）治疗。采用免疫组化方法，观察大鼠结肠黏膜组织IL-8、MCP-1蛋白的表达。结果模型组大鼠结肠黏膜组织MCP-1和IL-8蛋白表达阳性目标总面积和积分光密度均明显高于正常对照组（P〈0．01），经隔药灸和温和灸治疗后，CD大鼠结肠黏膜组织MCP-1和IL-8蛋白表达阳性目标总面积和积分光密度均显著降低（P〈0．01）。结论隔药灸和温和灸能够下调CD大鼠结肠黏膜MCP-1和IL-8蛋白的表达。     

艾灸预处理“天枢”穴对溃疡性结肠炎大鼠肠黏膜屏障保护的机制研究

目的：观察艾灸预处理“天枢”穴对UC大鼠的抗炎效应,探讨艾灸预处理“天枢”穴对UC大鼠肠黏膜屏障的保护机制。方法：SD雄性大鼠28只,随机分为正常组、模型组、隔药灸预处理组和温和灸预处理组,每组7只。造模前先予隔药灸、温和灸预处理干预7天;预处理后予模型组、隔药灸预处理组、温和灸预处理组4%葡聚糖硫酸钠水溶液连续饮用7天。采用HE染色法观察大鼠结肠组织病理学变化;免疫组化法和western blot法检测大鼠结肠组织分子的蛋白表达。结果：与正常组比较,模型组结肠组织病理学评分显著升高,Occludin、JAM1、MUC2、ZO-1蛋白表达显著降低,NF-κBp65、IL-1β蛋白表达显著升高（P ＜ 0.05）;与模型组比较,隔药灸预处理组结肠组织病理学评分显著降低,Occludin、MUC2、JAM1、ZO-1蛋白表达显著升高,NF-κBp65、IL-1β蛋白表达显著降低（P ＜ 0.05）,温和灸预处理组结肠组织病理学评分显著降低,Occludin、MUC2、ZO-1蛋白表达显著升高,IL-1β蛋白表达显著降低（P ＜ 0.05）。结论：隔药灸预处理、温和灸预处理“天枢”穴均能升高UC大鼠肠黏膜屏障相关蛋白的表达,可能是艾灸有效预防减轻UC的机制之一。     

艾灸对克罗恩病大鼠结肠NF-κB p65及TNF-α、IL-1β调节作用的研究

目的：通过观察隔药灸对克罗恩病大鼠结肠NF-κB p65及炎症因子TNF-α、IL-1β表达的影响,探讨艾灸治疗克罗恩病的效应机制。方法：清洁级雄性SD大鼠随机分为4组,分别为正常组、模型组、隔药灸组和西药组,采用三硝基苯磺酸（TN-BS）合乙醇溶液灌肠制备克罗恩病大鼠模型,模型制备成功后,隔药灸组取天枢穴（双）、气海穴进行隔药灸治疗,西药组采用美沙拉秦灌胃治疗。治疗结束后,采用HE染色,光镜下观察各组大鼠结肠组织病理学变化,应用免疫组织化学技术观察各组大鼠结肠NF-κB p65与TNF-α、IL-1β的表达,并进行图像分析。结果：与正常组比较,模型组大鼠结肠组织呈现裂隙状溃疡及炎症表现,部分大鼠结肠在黏膜下层可见肉芽肿形成,结肠NF-κB p65、TNF-α、IL-1β的表达均显著增高（均P〈0.01）;与模型组比较,隔药灸组、西药组大鼠结肠形态结构改善,肠道炎症减轻,结肠NF-κB p65与TNF-α、IL-1β的表达均显著降低（均P〈0.01）;相关性分析结果显示,TNF-α、IL-1β的表达与NF-κB p65的表达均呈正相关性。结论：隔药灸能下调克罗恩病大鼠结肠NF-κB p65、TNF-α、IL-1β的表达;艾灸（隔药灸）可能通过抑制克罗恩病大鼠结肠NF-κB p65的表达,进而减少其下游炎症因子TNF-α、IL-1β的表达,从而减轻肠道炎症,改善结肠组织形态结构,发挥治疗作用。     

艾灸预处理天枢穴对溃疡性结肠炎大鼠结肠TLR4/TRIF信号通路调节作用的研究

目的观察艾灸预处理天枢穴对溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠TLR4/TRIF信号通路的影响,探讨艾灸预处理防治UC的作用机制.方法将28只SD雄性大鼠随机分为正常组、模型组、隔药灸预处理组和温和灸预处理组.采用4%葡聚糖硫酸钠水溶液连续饮用7 d制备UC模型.隔药灸预处理组、温和灸预处理组先干预7 d后,模型组、隔药灸预处理组、温和灸预处理组再制备UC模型.采用HE染色法,观察大鼠结肠组织病理学变化;应用免疫组化、免疫印记技术检测大鼠结肠TLR4、TRIF、TRAF6、IRF3、IKK的蛋白表达.结果模型组大鼠结肠黏膜结构损伤较正常组明显加重;隔药灸预处理组和温和灸预处理组,结肠黏膜结构损伤较模型组明显改善.与正常组比较,模型组结肠TLR4、TRIF、TRAF6、IRF3、IKK蛋白表达均显著升高(P<0.05);与模型组比较,隔药灸预处理组、温和灸预处理组结肠TLR4、TRIF、TRAF6、IRF3蛋白表达均显著降低(P<0.05),隔药灸预处理组结肠IKK蛋白表达显著降低(P<0.05).结论艾灸预处理天枢穴能下调UC大鼠结肠TLR4、TRIF、TRAF6、IRF3蛋白表达;下调TLR4/TRIF信号通路相关分子的蛋白表达可能是艾灸预处理防治UC的作用机制之一.     

艾灸对溃疡性结肠炎肠纤维化大鼠结肠成纤维细胞增殖影响的研究

目的 从对溃疡性结肠炎大鼠结肠成纤维细胞增殖影响的角度,探讨艾灸防治溃疡性结肠炎肠纤维化的作用机制.方法 采用免疫法加局部刺激制备溃疡性结肠炎肠纤维化大鼠模型,随机将大鼠分为正常组、模型组、隔药灸组、温和灸组和西药组.隔药灸组和温和灸组选取天枢、气海穴分别进行隔药灸和温和灸治疗,西药组柳氮磺胺吡啶溶液灌胃治疗,治疗结束后处死大鼠,剖取结肠组织,分离并培养结肠成纤维细胞,流式细胞术观察各组大鼠结肠成纤维细胞增殖情况.结果 模型大鼠结肠成纤维细胞增殖率增高(P<0.01),处于G0-G1期细胞减少(P<0.01),G2-M期和S期细胞增多(P<0.01),隔药灸、温和灸疗法能显著改善大鼠结肠成纤维细胞的增殖率(P<0.01),使处于G0-G1期细胞、G2-M期和S期细胞趋向正常.结论 UC大鼠CFB增殖加快.隔药灸和温和灸天枢、气海穴治疗能显著抑制大鼠CFB的过度增殖.     

艾灸对克罗恩病肠纤维化大鼠结肠黏膜TGF-β及Smad4表达的影响

目的:观察艾灸对克罗恩病(Crohn Disease,CD)肠纤维化大鼠结肠转化生长因子-β(Transforming Growth Factor-β,TGF-β)蛋白、Smad4蛋白及mRNA表达的影响,从TGF-β/Smads信号途径探讨针灸治疗CD肠纤维化的作用机制.方法:采用雄性清洁级SD大鼠建立CD肠纤维化模型,应用随机的方法,将大鼠分为正常组、模型组、温和灸组、电针组、隔药灸组.正常组和模型组不予任何治疗;温和灸组、电针组、隔药灸组均取天枢、气海穴,分别施以温和灸、电针、隔药灸治疗.治疗结束后,采用免疫组织化学法(Immunohistochemistry,IHC)检测各组大鼠结肠TGF-β与Smad4蛋白的表达;采用荧光定量聚合酶链反应法(Fluorescence Quantitative Polymerase Chain Reaction,FQ-PCR)检测结肠Smad4 mRNA的表达.结果:与正常组比较,模型组大鼠结肠TGF-β蛋白、Smad4蛋白及mRNA表达增加(P＜0.01).经温和灸、电针、隔药灸治疗后,与模型组比较,TGF-β蛋白、Smad4蛋白及mRNA表达均有不同程度的降低(P＜0.01或P＜0.05).结论:CD肠纤维化大鼠结肠TGF-β蛋白、Smad4蛋白及mRNA表达显著增多,温和灸、电针、隔药灸天枢和气海穴均可下调CD肠纤维化大鼠结肠TGF-β蛋白、Smad4蛋白及mRNA的异常表达.     

艾灸对克罗恩病肠纤维化大鼠结肠黏膜TGF-β及Smad4表达的影响

目的：观察艾灸对克罗恩病（Crohn Disease, CD）肠纤维化大鼠结肠转化生长因子-β （Transforming Growth Factor-β, TGF-β）蛋白、Smad4 蛋白及 mRNA 表达的影响,从 TGF-β/Smads 信号途径探讨针灸治疗CD肠纤维化的作用机制。方法：采用雄性清洁级 SD 大鼠建立 CD 肠纤维化模型, 应用随机的方法,将大鼠分为正常组、模型组、温和灸组、电针组、隔药灸组。正常组和模型组不予任何治疗;温和灸组、电针组、隔药灸组均取天枢、气海穴,分别施以温和灸、电针、隔药灸治疗。治疗结束后,采用免疫组织化学法（Immunohistochemistry, IHC）检测各组大鼠结肠 TGF-β与Smad4蛋白的表达;采用荧光定量聚合酶链反应法（Fluorescence Quantitative Polymerase Chain Reaction, FQ-PCR） 检测结肠 Smad4 mRNA 的表达。结果：与正常组比较,模型组大鼠结肠 TGF-β蛋白、Smad4 蛋白及 mRNA 表达增加（P〈0.01）。经温和灸、电针、隔药灸治疗后,与模型组比较,TGF-β蛋白、Smad4 蛋白及 mRNA 表达均有不同程度的降低（P〈0.01 或 P〈0.05）。结论：CD肠纤维化大鼠结肠TGF-β蛋白、 Smad4 蛋白及mRNA表达显著增多,温和灸、电针、隔药灸天枢和气海穴均可下调CD肠纤维化大鼠结肠TGF-β蛋白、Smad4 蛋白及 mRNA 的异常表达。     

隔药灸对TNF-αzjtnyx-e202101-NF-κB-MLCK介导克罗恩病肠上皮细胞紧密连接的影响

目的:探讨隔药灸对肿瘤坏死因子-α(TNF-α)-核因子κB(NF-κB)-肌球蛋白轻链激酶(MLCK)介导克罗恩病(CD)肠上皮细胞紧密连接的影响.方法:将48只雄性SD大鼠随机分成正常组(NC)、模型组(MC)、隔药灸组(HPM)和美沙拉嗪组(MESA),每组12只.采用2,4,6-三硝基苯磺酸(TNBS)制备CD...     

针灸对大鼠溃疡性结肠炎结肠上皮细胞凋亡影响的实验研究

目的:探讨针灸调节大鼠溃疡性结肠炎结肠上皮细胞凋亡的作用机制.方法:在建立大鼠溃疡性结肠炎模型的基础上,随机分为模型组、隔药灸组、电针组,并设立正常组,隔药灸组和电针组选取"气海"穴、"天枢"穴,分别进行隔药灸与电针治疗.疗程结束后,剖取动物结肠组织,应用电镜、流式细胞仪观察各组大鼠结肠组织结构的改变及上皮细胞凋亡的变化.结果:与正常组大鼠比较,溃疡性结肠炎模型大鼠在结肠组织病理学改变的同时上皮细胞凋亡大量增加,电针、隔药灸可使上皮细胞凋亡得到显著地抑制.结论:电针、隔药灸治疗溃疡性结肠炎的主要作用机制之一是调节溃疡性结肠炎结肠上皮细胞的凋亡异常.     

艾灸预处理对大鼠应激性胃黏膜损伤的保护作用

目的:探讨艾灸预处理对大鼠应激性胃黏膜损伤保护作用及抗炎症损伤机制。方法:将48只健康SD大鼠随机分为4组,即空白组、模型组、艾灸穴位组、艾灸非穴组。束缚冷应激法制作应激性胃黏膜损伤大鼠模型,按Guth法计算胃黏膜损伤指数(UI),光镜下观察大鼠胃黏膜组织学改变,放射免疫法测定血清IL-1β,TNF-α和IL-10的含量。结果:与艾灸非穴组比较,艾灸足三里、中脘等穴位可使应激性胃黏膜损伤大鼠UI明显下降(P0.01)、血清IL-1β含量降低(P0.05)、TNF-α含量降低(P0.01)、IL-10含量升高(P0.05)。结论:艾灸足三里、中脘等穴位预处理可促进束缚水浸应激所造成大鼠胃黏膜损伤的修复、减轻急性炎症反应,该保护作用可能是通过抑制细胞炎症反应的免疫促炎因子IL-1β、TNF-α和促进抗炎因子IL-10而达到其抗胃黏膜损伤作用。     

Effect of Moxibustion on the Synthesis and Secretion of Collagen by Colonic Fibroblasts in Ulcerative Colitis Fibrosis Rats

目的：从对溃疡性结肠炎大鼠结肠成纤维细胞（Colonic Fibroblast，CFB）合成分泌胶原的影响的角度，探讨艾灸防治溃疡性结肠炎肠纤维化的作用机制。方法：采用免疫法加局部刺激制备溃疡性结肠炎肠纤维化大鼠模型，随机将大鼠分为正常组、模型组、隔药灸组、温和灸组和西药组。隔药灸组、温和灸组选取天枢、气海穴分别进行隔药灸、温和灸治疗，西药组柳氮磺胺吡啶溶液灌胃治疗。分离并培养结肠成纤维细胞，ELISA法观察艾灸对溃疡性结肠炎肠纤维化大鼠结肠成纤维细胞合成分泌Ⅰ、Ⅲ、Ⅳ型胶原的影响。结果：UC纤维化大鼠CFB培养上清液能显著激发正常大鼠CFB大量分泌Ⅰ、Ⅲ、Ⅳ型胶原，隔药灸组、温和灸组上清液则显著抑制正常大鼠CFBⅠ、Ⅲ、Ⅳ型胶原的分泌，西药组上清液对Ⅰ、Ⅲ型胶原分泌有一定抑制作用。结论：艾灸能够调节溃疡性结肠炎肠纤维化大鼠结肠成纤维细胞合成分泌Ⅰ、Ⅲ、Ⅳ型胶原。     

麦粒灸对肺炎链球菌肺炎大鼠肺泡巨噬细胞的免疫调节作用

目的:观察麦粒灸对肺炎链球菌肺炎大鼠肺泡巨噬细胞(AM)的免疫调节作用。方法:将30只SD大鼠随机分为正常组、模型组、麦粒灸组,每组10只。除正常组外,其余组大鼠制备肺炎链球菌肺炎模型。造模成功后,麦粒灸组大鼠于大椎、关元及双侧足三里、肺俞穴行7 d麦粒灸治疗。观察各组大鼠一般情况,血常规检测外周血白细胞计数,ELISA检测血清肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)含量,HE染色观察肺组织病理学变化,中性红法检测AM吞噬功能,qRT-PCR及Western blot分别检测M1、M2型AM标记物CD86、CD206 mRNA及蛋白表达。结果:与正常组比较,模型组大鼠精神懈怠,皮毛晦暗,外周血白细胞计数和血清TNF-α、IL-10含量均升高,肺组织出现病理损伤,AM吞噬OD值降低,CD86 mRNA及蛋白水平升高,CD206 mRNA及蛋白水平降低(P<0.05)。与模型组比较,麦粒灸组大鼠精神及皮毛状态改善,外周血白细胞计数降低,血清TNF-α、IL-10含量降低,肺组织病理损伤改善,AM吞噬OD值升高,CD86 mRNA及蛋白水平降低,CD206 mRNA及...     

Regulatory effect of moxibustion for rats with ulcerative colitis on the macrophage functional phenotype protein of lung tissue

Objective

To observe the regulatory effect of moxibustion on the expression of CD86 and CD163, which are the important functional phenotypes of macrophage differentiation in lung tissue of ulcerative colitis (UC) rats. The mechanism of the regulatory effect of moxibustion for macrophage differentiation based on the key cytokine interferon-gamma (IFN-γ), tumor necrosis death factor-alpha (TNF-α), interleukin- 4 (IL-4) and interleukin-13 (IL-13) was also explored.

Methods

Forty SD rats were randomly divided into a normal group (NG), a model group (MG), a normal moxibustion group (NMG) and a smokeless moxibustion group (SMG). The model of UC was made by antigen immunization combined with enema with topical formalin. The rats in the normal moxibustion group accepted moxibustion at bilateral Tianshu (ST 25), while the rats in the smokeless moxibustion group with smokeless moxibustion at bilateral Tianshu (ST 25), each 10 min, once a day for 8 d. After treatments, the hematoxylin-eosin (HE) staining was used to observe the pathological changes of colonic tissue, the Western blotting (WB) was used to observe the expression of CD86 and CD163, two important functional phenotypes of macrophage differentiation in lung tissue of UC rats, while the enzyme-linked immunosorbent assay (ELISA) was used to assess the content of IFN-γ, TNF-α, IL-4 and IL-13, the key cytokines of macrophage differentiation in micro environment of the lung tissue.

Results

Compared with the NG, the colons of rats in MG were injured more seriously, and the scores of gross observation and histological examination were significantly higher (P<0.05). Compared with the MG, the pathological changes of the two groups of rats with moxibustion treatment were improved, which presented with ulcer repair, inflammation dissipation, and the general score and histological score of the two groups were decreased (P<0.05). Compared with the NG, the expression of CD86 in the lung tissue of rats in the MG was increased (P<0.05), and CD163 expression was decreased (P<0.05). Compared with the MG and the SMG, the expression of CD86 in the lung tissue of the rats in the NMG was significantly lower than those in the MG and SMG, and CD163 was higher (P<0.05), while the differences were not statistically significant between the MG and the SMG (P>0.05). Compared with the NG, the expression of key cytokines in lung tissue of MG was abnormal, the contents of IFN-γ and TNF-α increased (P<0.05), while the IL-4 and IL-13 decreased (P<0.05). The IL-4 and IL-13 were significantly increased in lung tissue of rats in the NMG (P<0.05), and the IFN-γ and TNF-α were reduced (P<0.05), compared with those in the MG and the SMG, while the differences were not statistically significant between the MG and the SMG (P>0.05).

Conclusion

Moxibustion can increase the expression of CD163, the important functional phenotype of macrophages in lung tissue of UC rats, and the differentiation critical cytokines IL-4 and IL-13. It can also reduce activated phenotype CD86 and its differentiation critical cytokines IFN-γ and TNF-α in the lung tissue of UC rats.

     

穴位埋线对实验性结肠炎大鼠肠黏膜上皮屏障的影响

目的:观察穴位埋线对"虚、瘀"状态下溃疡性结肠炎(UC)模型大鼠肠黏膜上皮屏障的影响,探讨其改善UC的作用机制。方法:雄性SD大鼠随机分为正常组12只与造模组48只。造模组采用腺嘌呤、番泻叶分阶段灌胃制造大鼠"虚、瘀"状态,接着用5%2,4,6-三硝基苯磺酸(TNBS,100 mg/kg)加0.25 mL 50%乙醇灌肠建立UC大鼠模型。将造模成功的大鼠随机分为模型组、柳氮磺吡啶组和穴位埋线组,每组10只。穴位埋线组予以双侧"天枢""足三里""膈俞""脾俞""肾俞""大肠俞"埋线治疗,14 d埋线1次,共3次。柳氮磺吡啶组予以柳氮磺吡啶灌胃,每日1次,共42 d。观察大鼠一般状态及体质量变化,采用肉眼及HE染色观察大鼠结肠组织外观及病理变化,ELISA法检测各组大鼠血清中D乳酸(D-LA)、二胺氧化酶(DAO)和蛋白激酶C(PKC)含量,荧光定量PCR法检测各组大鼠结肠组织中肠黏膜紧密连接蛋白occludin、 claudin8、cingulin、zonulin的mRNA水平,Western blot法检测各组大鼠结肠组织中occludin、claudin8、cingulin、 zonulin和PKC的蛋白表达水平。结果:与正常组比较,模型组大鼠体质量显著降低(P<0.01),结肠组织明显肿胀,肠壁充血,结肠黏膜大量炎性细胞浸润,结肠黏膜损伤指数评分显著升高(P<0.01),血清中D-LA、DAO和PKC含量显著升高(P<0.01),结肠组织内occludin、claudin8、cingulin mRNA和蛋白表达水平均显著降低(P<0.01),zonulin mRNA和蛋白表达水平显著升高(P<0.01),PKC蛋白表达水平显著升高(P<0.01)。与模型组比较,柳氮磺吡啶组和穴位埋线组大鼠体质量显著升高(P<0.01),结肠病变明显减轻,黏膜损伤指数评分显著降低(P<0.05),血清中D-LA、DAO和PKC含量显著降低(P<0.01),结肠组织内occludin、claudin8、cingulin mRNA和蛋白表达水平显著增高(P<0.05,P<0.01),zonulin mRNA和蛋白表达水平显著降低(P<0.01),PKC蛋白表达水平显著降低(P<0.01)。穴位埋线组和柳氮磺吡啶组各指标比较差异无统计学意义(P>0.05)。结论:穴位埋线可调节上皮紧密连接,修复肠黏膜上皮屏障,促进肠黏膜愈合,其作用机制可能与抑制PKC的激活有关。     

Experimental study on the effect of different moxibustion durations on rats with rheumatoid arthritis

Objective

To observe the effect of different moxibustion durations on rats with rheumatoid arthritis (RA) and to evaluate the relationship between moxibustion amount and moxibustion efficacy.

Methods

Eight rats were randomly selected as a normal group from the 40 male Sprague-Dawley (SD) rats, and the other 32 rats were used to establish type II collagen-induced RA models. After successful modeling, the 32 rats were randomly divided into a model group, a moxibustion for 20 min group, a moxibustion for 40 min group and a moxibustion for 60 min group, with 8 rats in each group. Rats in the normal group did not receive modeling and moxibustion intervention; rats in the model group did not receive moxibustion after modeling; rats in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group were treated with moxibustion at Shenshu (BL 23) and Zusanli (ST 36) for 20 min, 40 min and 60 min, respectively. Six days were a course of treatment, with a total of 3-course treatments and a 1-day rest between the courses of treatment. After treatment, the serum levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α, arthritis index (AI) scores, toe volumes and pathological score of synovitis were evaluated in the rats.

Results

Compared with the normal group, the serum IL-1β and TNF-α levels, and the toe volumes in the model group were increased, and the differences were statistically significant (P<0.01), before the treatment. Compared with the model group, the serum IL-1β and TNF-α levels, toe volumes and arthritis index (AI) scores were significantly decreased in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group (P<0.05 or P<0.01 ). Compared with the moxibustion for 20 min group and the moxibustion for 60 min group, serum IL-1β and TNF-α levels, toe volumes and AI scores were decreased more significantly in moxibustion for 40 min group, and the differences were statistically significant (P<0.05 or P<0.01). There were no significant differences in serum IL-1β and TNF-α levels, AI scores and toe volumes between the moxibustion for 20 min group and the moxibustion for 60 min group (all P>0.05). The synovial histopathological improvement was the most obvious in the moxibustion for 40 min group, when the synovial histopathological changes were compared among the moxibustion for 20 min group, moxibustion for 40 min group and moxibustion for 60 min group.

Conclusion

The therapeutic efficacy of moxibustion for 40 min in RA rats was more significant than that of moxibustion for 20 min and moxibustion for 60 min, indicating that the duration of moxibustion is the main factor affecting its therapeutic efficacy.