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
6期
758-764,781
,共8页
邵珲%王进海%张杰%杨龙宝%谢丹红
邵琿%王進海%張傑%楊龍寶%謝丹紅
소혼%왕진해%장걸%양룡보%사단홍
肠道菌群失调%内脏高敏感%肠易激综合征%低度炎症%紧密连接蛋白(ZO-1)%Toll 样 4 受体(TLR4)%益生菌
腸道菌群失調%內髒高敏感%腸易激綜閤徵%低度炎癥%緊密連接蛋白(ZO-1)%Toll 樣 4 受體(TLR4)%益生菌
장도균군실조%내장고민감%장역격종합정%저도염증%긴밀련접단백(ZO-1)%Toll 양 4 수체(TLR4)%익생균
intestinal dysbacteriosis%visceral hypersensitivity%irritable bowel syndrome%low-grade inflamma-tion%tight junction protein (ZO-1)%Toll-like receptor 4 (TLR4)%probiotics
目的：检测肠道菌群失调大鼠的内脏敏感性变化及紧密连接蛋白(ZO-1)、Toll 样4受体(TLR4)的表达,探讨肠道菌群失调对内脏敏感性的影响及其机制。方法SPF 级 SD 大鼠30只,随机分为正常对照组(12只)、菌群失调组(18只)。菌群失调组采用盐酸林可霉素(300 mg/mL)灌胃,1 mL/(次·只),1次/d,连续给药7 d；正常对照组采用等量生理盐水灌胃。第8天随机抽取菌群失调组和正常对照组大鼠各6只,检测模型是否成功。模型建立成功后,将其余12只菌群失调组大鼠随机分组：阴性对照组、益生菌干预组,每组6只。益生菌干预组给予双歧杆菌三联活菌胶囊(培菲康)灌胃,1粒/3 mL 生理盐水,1 mL/(次·只),1次/d,连续给药7 d；阴性对照组给予同等量生理盐水灌胃。第8天进行粪便菌群培养、检测内脏敏感性、检测结肠组织中 ZO-1及 TLR4的 mRNA 和蛋白表达、检测血清中炎性因子 IL-10、TNFα的表达。结果与正常对照组比较,菌群失调组大鼠结肠 ZO-1的 mRNA 含量及蛋白表达显著降低,而结肠 TLR4的含量及蛋白表达显著升高,且血清中促炎因子 TNFα表达显著升高,而抑炎因子 IL-10的表达显著降低(P <0.05)。与菌群失调组相比,益生菌干预组结肠 ZO-1的 mRNA 含量及蛋白表达显著升高,而结肠 TLR4的 mRNA 含量及蛋白表达显著降低,且血清中促炎因子 TNFα表达显著降低而抑炎因子 IL-10的表达显著升高(P <0.05)。结论通过抑制结肠中 ZO-1的表达及升高 TLR4的表达,进而导致慢性低度炎症的发生可能是肠道菌群失调导致内脏高敏感的作用机制之一,同时益生菌制剂可能通过恢复肠道菌群变化进而改善内脏高敏感。
目的：檢測腸道菌群失調大鼠的內髒敏感性變化及緊密連接蛋白(ZO-1)、Toll 樣4受體(TLR4)的錶達,探討腸道菌群失調對內髒敏感性的影響及其機製。方法SPF 級 SD 大鼠30隻,隨機分為正常對照組(12隻)、菌群失調組(18隻)。菌群失調組採用鹽痠林可黴素(300 mg/mL)灌胃,1 mL/(次·隻),1次/d,連續給藥7 d；正常對照組採用等量生理鹽水灌胃。第8天隨機抽取菌群失調組和正常對照組大鼠各6隻,檢測模型是否成功。模型建立成功後,將其餘12隻菌群失調組大鼠隨機分組：陰性對照組、益生菌榦預組,每組6隻。益生菌榦預組給予雙歧桿菌三聯活菌膠囊(培菲康)灌胃,1粒/3 mL 生理鹽水,1 mL/(次·隻),1次/d,連續給藥7 d；陰性對照組給予同等量生理鹽水灌胃。第8天進行糞便菌群培養、檢測內髒敏感性、檢測結腸組織中 ZO-1及 TLR4的 mRNA 和蛋白錶達、檢測血清中炎性因子 IL-10、TNFα的錶達。結果與正常對照組比較,菌群失調組大鼠結腸 ZO-1的 mRNA 含量及蛋白錶達顯著降低,而結腸 TLR4的含量及蛋白錶達顯著升高,且血清中促炎因子 TNFα錶達顯著升高,而抑炎因子 IL-10的錶達顯著降低(P <0.05)。與菌群失調組相比,益生菌榦預組結腸 ZO-1的 mRNA 含量及蛋白錶達顯著升高,而結腸 TLR4的 mRNA 含量及蛋白錶達顯著降低,且血清中促炎因子 TNFα錶達顯著降低而抑炎因子 IL-10的錶達顯著升高(P <0.05)。結論通過抑製結腸中 ZO-1的錶達及升高 TLR4的錶達,進而導緻慢性低度炎癥的髮生可能是腸道菌群失調導緻內髒高敏感的作用機製之一,同時益生菌製劑可能通過恢複腸道菌群變化進而改善內髒高敏感。
목적：검측장도균군실조대서적내장민감성변화급긴밀련접단백(ZO-1)、Toll 양4수체(TLR4)적표체,탐토장도균군실조대내장민감성적영향급기궤제。방법SPF 급 SD 대서30지,수궤분위정상대조조(12지)、균군실조조(18지)。균군실조조채용염산림가매소(300 mg/mL)관위,1 mL/(차·지),1차/d,련속급약7 d；정상대조조채용등량생리염수관위。제8천수궤추취균군실조조화정상대조조대서각6지,검측모형시부성공。모형건립성공후,장기여12지균군실조조대서수궤분조：음성대조조、익생균간예조,매조6지。익생균간예조급여쌍기간균삼련활균효낭(배비강)관위,1립/3 mL 생리염수,1 mL/(차·지),1차/d,련속급약7 d；음성대조조급여동등량생리염수관위。제8천진행분편균군배양、검측내장민감성、검측결장조직중 ZO-1급 TLR4적 mRNA 화단백표체、검측혈청중염성인자 IL-10、TNFα적표체。결과여정상대조조비교,균군실조조대서결장 ZO-1적 mRNA 함량급단백표체현저강저,이결장 TLR4적함량급단백표체현저승고,차혈청중촉염인자 TNFα표체현저승고,이억염인자 IL-10적표체현저강저(P <0.05)。여균군실조조상비,익생균간예조결장 ZO-1적 mRNA 함량급단백표체현저승고,이결장 TLR4적 mRNA 함량급단백표체현저강저,차혈청중촉염인자 TNFα표체현저강저이억염인자 IL-10적표체현저승고(P <0.05)。결론통과억제결장중 ZO-1적표체급승고 TLR4적표체,진이도치만성저도염증적발생가능시장도균군실조도치내장고민감적작용궤제지일,동시익생균제제가능통과회복장도균군변화진이개선내장고민감。
Objective To detect the changes of visceral sensitivity in rats presenting intestinal dysbacteriosis and the expressions of tight junction protein (ZO-1)and Toll-like receptor 4 (TLR4)so as to explore the effect of intestinal dysbacteriosis on visceral sensitivity and the possible mechanisms.Methods We randomly divided 30 male SD rats of SPF grade into normal control group (n = 12 )and dysbacteriosis group (n = 18 ).Rats in dysbacteriosis group were administered with lincomycin hydrochloride (300 mg/mL),1 mL each time per rat once a day for 7 consecutive days;those in normal control group were fed with the same amount of saline.On the eighth day,six rats were randomly selected from normal control group and dysbacteriosis group respectively to detect whether the model was successful.After the model was successfully constructed,the remaining 12 dysbacteriosis rats were randomly divided into the negative control group and the probiotics intervention group with 6 in each.Rats in the intervention group were given probiotic bifidobacterium triple viable capsules (Bifico)orally,one capsule with 1/3 mL of saline,1 mL each time per rat once a day for 7 consecutive days;those in the negative control group received the same amount of saline.On the eighth day,fresh feces was cultured for flora to detect visceral sensitivity by abdominal withdrawal reflex (AWR),the mRNA and protein expressions of ZO-1 and TLR4 in the colon,and the expression of serum inflammatory cytokines IL-10 and TNFα.Results The expression of ZO-1 in the colon was significantly lower in the rats of dysbacteriosis group than those in the control group,and the expression of TLR4 was also significantly increased.Correspondingly,the expression of pro-inflammatory factor TNFα in the serum of the rats in dysbacteriosis group was significantly increased,while that of anti-inflammatory factor IL-10 was significantly lower than in the control group (P <0.05).Furthermore,compared with dysbacteriosis group,the expression of ZO-1 was increased significantly and TLR4 was decreased in probiotics group in varying degrees. Similarly,the expression of TNFα was obviously lower while that of IL-10 in the serum was higher (P < 0.05 ). Conclusion Inhibiting the expression of ZO-1 and increasing the expression of TLR4,thus leading to chronic low-grade inflammation, may be one mechanism of visceral hypersensitivity caused by intestinal dysbacteriosis. Probiotics may restore the dysbacteriosis and thus improve visceral hypersensitivity.
