西安交通大学学报（医学版）
西安交通大學學報（醫學版）
서안교통대학학보（의학판）
JOURNAL OF XI'AN JIAOTONG UNIVERSITY(MEDICAL SCIENCES)
2015年
2期
263-270
,共8页
电针%溃疡性结肠炎%toll 样受体 4(TLR4)NF-κB 信号通路%PI3K/AKT/NF-κB 信号通路%活化核转录因子-κB%肿瘤坏死因子-αmRNA%IL-1βmRNA
電針%潰瘍性結腸炎%toll 樣受體 4(TLR4)NF-κB 信號通路%PI3K/AKT/NF-κB 信號通路%活化覈轉錄因子-κB%腫瘤壞死因子-αmRNA%IL-1βmRNA
전침%궤양성결장염%toll 양수체 4(TLR4)NF-κB 신호통로%PI3K/AKT/NF-κB 신호통로%활화핵전록인자-κB%종류배사인자-αmRNA%IL-1βmRNA
electroacupuncture%ulcerative colitis%toll-like receptor 4 (TLR4 )/NF-ΚB signaling pathway%PI3K/AKT/NF-κB signaling pathway%active NF-κB%TNF-αmRNA%IL-1βmRNA
目的：探讨三硝基苯磺酸(TNBS)/乙醇灌肠液诱导大鼠溃疡性结肠炎 TLR4/NF-κB 和 PI3K/AKT /NF-κB 信号通路的表达及电针的干预作用。方法雄性 Wistar 大鼠240只,随机分为正常对照组、模型组、电针组、TLR4单克隆抗体(TLR4mAb)组、LY294002组和 TLR4mAb-LY294002联合治疗(T&L)组。以 TNBS/乙醇溶液灌肠诱导大鼠溃疡性结肠炎模型,电针组造模同时给予电针足三里穴治疗,TLR4mAb 组给予 TLR4mAb 腹腔注射,LY294002组则予 LY294002注射液腹腔注射,T&L 组则同时给与上述 TLR4mAb 和 LY294002腹腔注射,共4周,每日行疾病活动指数(DAI)评分。造模4周后处死大鼠取结肠组织标本,评价结肠黏膜损伤指数(CMDI)和组织学损伤指数(TDI)；免疫印迹法测定结肠黏膜磷酸化 AKT(P-AKT)和活化 NF-κB 含量；RT-PCR 法检测结肠组织中 TLR4 mR-NA、PI3K mRNA、AKT mRNA、NF-κB mRNA、TNF-αmRNA 及 IL-1βmRNA 表达。结果与正常对照组比较,模型组结肠组织 TLR4 mRNA、PI3K mRNA、P-AKT、活化 NF-κB 、TNF-αmRNA、IL-1βmRNA 表达和 DAI、CMDI 及TDI 均显著升高(P <0.01)；与模型组比较,TLR4mAb 组 TLR4 mRNA 表达显著下降(P <0.01),LY294002组 PI3K mRNA、P-AKT 表达显著下降(P <0.01),而在电针组和 T&L 组 TLR4 mRNA、PI3K mRNA 及 P-AKT 表达均明显降低(P <0.01)；与上述变化相对应,与模型组相比,各治疗组活化 NF-κB、TNF-αmRNA、IL-1βmRNA 表达和 DAI、CMDI 及 TDI 均明显降低(P <0.05或 P <0.01),电针组和 T&L 组上述6指标优于 TLR4mAb 组和 LY294002组,活化 NF-κB 与 TNF-αmRNA 及 IL-1βmRNA 表达呈正相关(r 1=0.579,P <0.05；r 2=0.561,P <0.05)。结论电针足三里穴能显著降低实验性溃疡性结肠炎大鼠的 NF-κB 活性及 TNF-αmRNA、IL-1βmRNA 表达,从而减轻结肠组织损害程度,这与其阻断 TLR4/NF-κB 和 PI3K/AKT/NF-κB 信号通路有密切关系。
目的：探討三硝基苯磺痠(TNBS)/乙醇灌腸液誘導大鼠潰瘍性結腸炎 TLR4/NF-κB 和 PI3K/AKT /NF-κB 信號通路的錶達及電針的榦預作用。方法雄性 Wistar 大鼠240隻,隨機分為正常對照組、模型組、電針組、TLR4單剋隆抗體(TLR4mAb)組、LY294002組和 TLR4mAb-LY294002聯閤治療(T&L)組。以 TNBS/乙醇溶液灌腸誘導大鼠潰瘍性結腸炎模型,電針組造模同時給予電針足三裏穴治療,TLR4mAb 組給予 TLR4mAb 腹腔註射,LY294002組則予 LY294002註射液腹腔註射,T&L 組則同時給與上述 TLR4mAb 和 LY294002腹腔註射,共4週,每日行疾病活動指數(DAI)評分。造模4週後處死大鼠取結腸組織標本,評價結腸黏膜損傷指數(CMDI)和組織學損傷指數(TDI)；免疫印跡法測定結腸黏膜燐痠化 AKT(P-AKT)和活化 NF-κB 含量；RT-PCR 法檢測結腸組織中 TLR4 mR-NA、PI3K mRNA、AKT mRNA、NF-κB mRNA、TNF-αmRNA 及 IL-1βmRNA 錶達。結果與正常對照組比較,模型組結腸組織 TLR4 mRNA、PI3K mRNA、P-AKT、活化 NF-κB 、TNF-αmRNA、IL-1βmRNA 錶達和 DAI、CMDI 及TDI 均顯著升高(P <0.01)；與模型組比較,TLR4mAb 組 TLR4 mRNA 錶達顯著下降(P <0.01),LY294002組 PI3K mRNA、P-AKT 錶達顯著下降(P <0.01),而在電針組和 T&L 組 TLR4 mRNA、PI3K mRNA 及 P-AKT 錶達均明顯降低(P <0.01)；與上述變化相對應,與模型組相比,各治療組活化 NF-κB、TNF-αmRNA、IL-1βmRNA 錶達和 DAI、CMDI 及 TDI 均明顯降低(P <0.05或 P <0.01),電針組和 T&L 組上述6指標優于 TLR4mAb 組和 LY294002組,活化 NF-κB 與 TNF-αmRNA 及 IL-1βmRNA 錶達呈正相關(r 1=0.579,P <0.05；r 2=0.561,P <0.05)。結論電針足三裏穴能顯著降低實驗性潰瘍性結腸炎大鼠的 NF-κB 活性及 TNF-αmRNA、IL-1βmRNA 錶達,從而減輕結腸組織損害程度,這與其阻斷 TLR4/NF-κB 和 PI3K/AKT/NF-κB 信號通路有密切關繫。
목적：탐토삼초기분광산(TNBS)/을순관장액유도대서궤양성결장염 TLR4/NF-κB 화 PI3K/AKT /NF-κB 신호통로적표체급전침적간예작용。방법웅성 Wistar 대서240지,수궤분위정상대조조、모형조、전침조、TLR4단극륭항체(TLR4mAb)조、LY294002조화 TLR4mAb-LY294002연합치료(T&L)조。이 TNBS/을순용액관장유도대서궤양성결장염모형,전침조조모동시급여전침족삼리혈치료,TLR4mAb 조급여 TLR4mAb 복강주사,LY294002조칙여 LY294002주사액복강주사,T&L 조칙동시급여상술 TLR4mAb 화 LY294002복강주사,공4주,매일행질병활동지수(DAI)평분。조모4주후처사대서취결장조직표본,평개결장점막손상지수(CMDI)화조직학손상지수(TDI)；면역인적법측정결장점막린산화 AKT(P-AKT)화활화 NF-κB 함량；RT-PCR 법검측결장조직중 TLR4 mR-NA、PI3K mRNA、AKT mRNA、NF-κB mRNA、TNF-αmRNA 급 IL-1βmRNA 표체。결과여정상대조조비교,모형조결장조직 TLR4 mRNA、PI3K mRNA、P-AKT、활화 NF-κB 、TNF-αmRNA、IL-1βmRNA 표체화 DAI、CMDI 급TDI 균현저승고(P <0.01)；여모형조비교,TLR4mAb 조 TLR4 mRNA 표체현저하강(P <0.01),LY294002조 PI3K mRNA、P-AKT 표체현저하강(P <0.01),이재전침조화 T&L 조 TLR4 mRNA、PI3K mRNA 급 P-AKT 표체균명현강저(P <0.01)；여상술변화상대응,여모형조상비,각치료조활화 NF-κB、TNF-αmRNA、IL-1βmRNA 표체화 DAI、CMDI 급 TDI 균명현강저(P <0.05혹 P <0.01),전침조화 T&L 조상술6지표우우 TLR4mAb 조화 LY294002조,활화 NF-κB 여 TNF-αmRNA 급 IL-1βmRNA 표체정정상관(r 1=0.579,P <0.05；r 2=0.561,P <0.05)。결론전침족삼리혈능현저강저실험성궤양성결장염대서적 NF-κB 활성급 TNF-αmRNA、IL-1βmRNA 표체,종이감경결장조직손해정도,저여기조단 TLR4/NF-κB 화 PI3K/AKT/NF-κB 신호통로유밀절관계。
ABSTRACT:Objective To explore the expressions of Toll-like receptor4/NF-κB and PI3K/AKT/NF-κB signa-ling pathways in rat ulcerative colitis (UC)induced by the combined enema of trinitrobenzene sulphonic acid and ethano and the interventional effect of electroacupuncture on them.Methods Totally 240 male Wistar rats were randomly divided into 6 groups:normal control group,model control group,electroacupuncture group,TLR4mAb group,LY294002 group,and TLR4mAb combined with LY294002 (T&L)group.The combined enema of trinitro-benzene sulphonic acid (TNB)and ethanol was intrarectally administered for 4 weeks to induce UC.At the same time of modeling ,Zusanli point was electro-acupunctured in electroacupuncture group while intraperitoneal injec-tion of TLR4mAb and LY294002 was given respectively to the corresponding group.Each rat was treated with the above-mentioned TLR4mAb injection and LY294002 injection in T&L group for 4 weeks.The disease activity index (DAI)of all the rats was evaluated daily.The rats were killed after 4 weeks.The colonic mucosa damage index (CMDI)and tissue damage index (TDI)were evaluated by a pathologic grading system.The expressions of P-Akt and active NF-κB protein in the colon mucosa were determined by Western blotting.TLR4 mRNA,PI3K mRNA, AKT mRNA,NF-κB mRNA,TNF-αmRNA and IL-1βmRNA expressions were measured with RT-PCR.Results Compared with those in normal control group,TLR4 mRNA,PI3K mRNA,P-AKT,active NF-κB,TNF-αmRNA and IL-1βmRNA expressions as well as DAI,CMDI and TDI were all increased obviously in model control group (P <0.01).Compared with those in model control group,TLR4mRNA expression was decreased obviously in TLR4mRNA group (P <0.01),the expressions of PI3KmRNA and P-AKT were decreased obviously in LY294002 group (P <0.01 ).Not only TLR4mRNA expression but also PI3KmRNA and P-AKT expressions were decreased significantly in electroacupuncture group and T&L group (P <0.01 ).Corresponding to the above-mentioned chan-ges,active NF-κB,TNF-αmRNA and IL-1βmRNA expressions as well as DAI,CMDI and TDI were decreased obvi-ously in all the treated groups compared with those in model control group (P <0.05 or P <0.01),but the six inde-xes were better in electroacupuncture group and T&L group than in TLR4mAb group and LY294002 group (P <0.05).There were obvious positive correlations of active NF-κB with TNF-αmRNA and IL-1β mRNA expressions (r 1 =0.579,P <0.05;r 2 =0.561,P <0.05).Conclusion Electroacupuncture can significantly decrease NF-κB activity and TNF-αmRNA and IL-1β mRNA expressions in UC rats,thus alleviating the severity of UC,which is closely correlated to its blocking both TLR4/NF-κB and PI3K/AKT/NF-κB signaling pathways.
