中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2012年
2期
118-123
,共6页
欧洋肖%陆允敏%秦环龙%陈红旗%朱梅影%陈维雄
歐洋肖%陸允敏%秦環龍%陳紅旂%硃梅影%陳維雄
구양초%륙윤민%진배룡%진홍기%주매영%진유웅
白细胞介素-10%纤维化%结肠%生物转化%上皮细胞%内质网
白細胞介素-10%纖維化%結腸%生物轉化%上皮細胞%內質網
백세포개소-10%섬유화%결장%생물전화%상피세포%내질망
Interleukin-10%Fibrosis%Colon%Biotransformation%Epithelial cells%Endoplasmic
目的 探讨白细胞介素(IL)-10对小鼠肠道纤维化及上皮-间质转化(EMT)的作用,以及该作用与内质网应激(ERS)的关系.方法 42只IL-10-/-小鼠分为纤维化模型组(n=18)、IL-10治疗组(n=12)和溶剂对照组(n=12),另取18只野生型小鼠作为阴性对照组.IL-10治疗组和溶剂对照组分别于第12周开始腹腔内注射IL-10和0.9％NaCl溶液,纤维化模型组和阴性对照组不予处理.采用HE染色及Masson胶原三色染色观察小鼠结肠组织损伤和纤维化变化,采用荧光定量PCR法检测小鼠结肠组织中胶原Ⅰ、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、平滑肌肌动蛋白(α-SMA)、E-细胞钙黏蛋白(E-cad)mRNA的表达.采用定量免疫组织化学染色检测小鼠结肠组织中α-SMA和E-cad的表达.结果 16周纤维化模型组、溶剂对照组结肠黏膜组织损伤评分(7.00±0.90和7.17±1.17)及胶原面积比[(17.78±4.15)％和(18.56±3.81)％]较阴性对照组[1.50±1.38和(9.11±2.99)％]和IL-10治疗组[4.33±0.82和(12.56±1.39)％]均明显升高(F=36.150,F=11.280；P=0.000).12、14、16周纤维化模型组、溶剂对照组GRP78、α-SMA、胶原Ⅰ表达量较同期阴性对照组均明显升高(均P＜0.05),E-cad表达量较阴性对照组明显降低(P＜0.05).12周纤维化模型组CHOP mRNA表达量[(0.95±0.12)％]较阴性对照组[(0.21±0.12)％]明显升高(t=5.188,P=0.000),但在14、16周各组之间差异无统计学意义(P＞0.05).14、16周IL-10治疗组GRP78、α-SMA、胶原Ⅰ表达量[14周:(0.73±0.31)％,(1.18.±0.11)％,(1.10±0.49)％；16周:(0.57±0.16)％,(0.81±0.50)％,(0.76±0.25)％]较纤维化模型组均明显降低(P＜0.05),E-cad表达量[14周:(0.73±0.29)％；16周:(0.97±0.25)％]较纤维化模型组[14周:(0.37±0.17)％；16周:(0.35±0.20)％]明显升高(F=6.524,P=0.003；,F=17.493,P=0.000).16周IL-10治疗组α-SMA表达量较溶剂对照组[(1.82±0.22)％]明显降低(F=9.842,P=0.000),E-cad表达量较溶剂对照组[(0.47±0.25)％]明显升高(F=17.493,P=0.000).结论 IL-10具有抑制EMT,减轻小鼠肠道纤维化的作用,可能与IL-10对ERS的调控有关.
目的 探討白細胞介素(IL)-10對小鼠腸道纖維化及上皮-間質轉化(EMT)的作用,以及該作用與內質網應激(ERS)的關繫.方法 42隻IL-10-/-小鼠分為纖維化模型組(n=18)、IL-10治療組(n=12)和溶劑對照組(n=12),另取18隻野生型小鼠作為陰性對照組.IL-10治療組和溶劑對照組分彆于第12週開始腹腔內註射IL-10和0.9％NaCl溶液,纖維化模型組和陰性對照組不予處理.採用HE染色及Masson膠原三色染色觀察小鼠結腸組織損傷和纖維化變化,採用熒光定量PCR法檢測小鼠結腸組織中膠原Ⅰ、葡萄糖調節蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、平滑肌肌動蛋白(α-SMA)、E-細胞鈣黏蛋白(E-cad)mRNA的錶達.採用定量免疫組織化學染色檢測小鼠結腸組織中α-SMA和E-cad的錶達.結果 16週纖維化模型組、溶劑對照組結腸黏膜組織損傷評分(7.00±0.90和7.17±1.17)及膠原麵積比[(17.78±4.15)％和(18.56±3.81)％]較陰性對照組[1.50±1.38和(9.11±2.99)％]和IL-10治療組[4.33±0.82和(12.56±1.39)％]均明顯升高(F=36.150,F=11.280；P=0.000).12、14、16週纖維化模型組、溶劑對照組GRP78、α-SMA、膠原Ⅰ錶達量較同期陰性對照組均明顯升高(均P＜0.05),E-cad錶達量較陰性對照組明顯降低(P＜0.05).12週纖維化模型組CHOP mRNA錶達量[(0.95±0.12)％]較陰性對照組[(0.21±0.12)％]明顯升高(t=5.188,P=0.000),但在14、16週各組之間差異無統計學意義(P＞0.05).14、16週IL-10治療組GRP78、α-SMA、膠原Ⅰ錶達量[14週:(0.73±0.31)％,(1.18.±0.11)％,(1.10±0.49)％；16週:(0.57±0.16)％,(0.81±0.50)％,(0.76±0.25)％]較纖維化模型組均明顯降低(P＜0.05),E-cad錶達量[14週:(0.73±0.29)％；16週:(0.97±0.25)％]較纖維化模型組[14週:(0.37±0.17)％；16週:(0.35±0.20)％]明顯升高(F=6.524,P=0.003；,F=17.493,P=0.000).16週IL-10治療組α-SMA錶達量較溶劑對照組[(1.82±0.22)％]明顯降低(F=9.842,P=0.000),E-cad錶達量較溶劑對照組[(0.47±0.25)％]明顯升高(F=17.493,P=0.000).結論 IL-10具有抑製EMT,減輕小鼠腸道纖維化的作用,可能與IL-10對ERS的調控有關.
목적 탐토백세포개소(IL)-10대소서장도섬유화급상피-간질전화(EMT)적작용,이급해작용여내질망응격(ERS)적관계.방법 42지IL-10-/-소서분위섬유화모형조(n=18)、IL-10치료조(n=12)화용제대조조(n=12),령취18지야생형소서작위음성대조조.IL-10치료조화용제대조조분별우제12주개시복강내주사IL-10화0.9％NaCl용액,섬유화모형조화음성대조조불여처리.채용HE염색급Masson효원삼색염색관찰소서결장조직손상화섬유화변화,채용형광정량PCR법검측소서결장조직중효원Ⅰ、포도당조절단백78(GRP78)、C/EBP동원단백(CHOP)、평활기기동단백(α-SMA)、E-세포개점단백(E-cad)mRNA적표체.채용정량면역조직화학염색검측소서결장조직중α-SMA화E-cad적표체.결과 16주섬유화모형조、용제대조조결장점막조직손상평분(7.00±0.90화7.17±1.17)급효원면적비[(17.78±4.15)％화(18.56±3.81)％]교음성대조조[1.50±1.38화(9.11±2.99)％]화IL-10치료조[4.33±0.82화(12.56±1.39)％]균명현승고(F=36.150,F=11.280；P=0.000).12、14、16주섬유화모형조、용제대조조GRP78、α-SMA、효원Ⅰ표체량교동기음성대조조균명현승고(균P＜0.05),E-cad표체량교음성대조조명현강저(P＜0.05).12주섬유화모형조CHOP mRNA표체량[(0.95±0.12)％]교음성대조조[(0.21±0.12)％]명현승고(t=5.188,P=0.000),단재14、16주각조지간차이무통계학의의(P＞0.05).14、16주IL-10치료조GRP78、α-SMA、효원Ⅰ표체량[14주:(0.73±0.31)％,(1.18.±0.11)％,(1.10±0.49)％；16주:(0.57±0.16)％,(0.81±0.50)％,(0.76±0.25)％]교섬유화모형조균명현강저(P＜0.05),E-cad표체량[14주:(0.73±0.29)％；16주:(0.97±0.25)％]교섬유화모형조[14주:(0.37±0.17)％；16주:(0.35±0.20)％]명현승고(F=6.524,P=0.003；,F=17.493,P=0.000).16주IL-10치료조α-SMA표체량교용제대조조[(1.82±0.22)％]명현강저(F=9.842,P=0.000),E-cad표체량교용제대조조[(0.47±0.25)％]명현승고(F=17.493,P=0.000).결론 IL-10구유억제EMT,감경소서장도섬유화적작용,가능여IL-10대ERS적조공유관.
Objective To investigate the effects of interleukin-10 on mice intestinal fibrosis and epithelial-mesenchymal transition(EMT),and the relation between these effects and endoplasmic reticulum stress(ERS).Methods Forty-two IL-10 knockout(IL-10-/-)mice were divided into fibrosis model group(n=18),IL-10 treatment group(n=12)and solvent control group(n =12),another 18 wild-type mice were taken as negative control group.IL-10 and 0.9％ NaCl were intraperitonealy injected in IL-10 treatment group and solvent control group respectively since 12th week,and mice were sacrificed at week 14th and 16th,and no treatment to fibrosis model group and negative control group.The injury and fibrosis in mice colon tissue were detected with HE staining and Masson collagen staining.The expressions of collagen Ⅰ,glucose-regulated protein78(GRP78),C/EBP homologous protein(CHOP)and a-smooth muscle actin(α-SMA)and E-cadherin(E-cad)at mRNA level were determined by realtime PCR.The expression of α-SMA and E-cad in mice colon tissue was examined by immunohistochemical staining.Results At 16th week,the colonic tissue injury scores(7.00±0.90,7.17±1.17)and collagen area ratio(17.78％±4.15％,18.56％±3.81％)of fibrosis model group and solvent control group significantly increased compared with negative control group(1.50±1.38 and 9.11％±2.99％)and IL-10 treatment qroup(4.33±0.82 and 12.56％±1.39％)(F=36.150,F=11.280； P=0.000).At week 12th,14th and 16th,the expressions of GRP78,α-SMA,collagen Ⅰ in fibrosis model group and solvent control group significantly increased compared with negative control group(all P＜0.05),however the expression of E-cad significantly decreased(P＜0.05).The expression of CHOP mRNA in fibrosis model group(0.95％ ±0.12％)significantly increased compared with negative control group(0.21％ ± 0.12％)at week 12th(t=5.188,P=0.000),however there was no statistical significant difference in groups at week 14th and 16th(P＞0.05).At week 14th and 16th,the expressions of GRP78,α-SMA and collagen Ⅰ(at week 14th:0.73％±0.31％,1.18％±0.11％ and 1.10％±0.49％； at week 16th:0.57％±0.16％,0.81％ ±0.50 ％ and 0.76 ％ ± 0.25 ％)in IL-10 treatment group were significantly lower than that of fibrosis model group(P＜0.05).The expression of E-cad(at week 14th:0.73％ ±0.29％ ； at week 16th:0.97％ ±0.25％)significantly increased compared with fibrosis model group(at week 14th:0.37％±0.17％； at week 16th:0.35％±0.20％)(F=6.524,P=0.003； F=17.493,P=0.000).However at week 16th,the expression of α-SMA in IL-10 treatment group was lower than that of solvent control group(1.82±0.22)％(F=9.842,P=0.000),and the expression of E-cad significantly increased than in solvent control group(0.47 ± 0.25)％(F=17.493,P =0.000).Conclusion IL-10 may have a role in inhibiting EMT and reducing intestinal fibrosis in mice,which may be related to the regulation of ERS by IL-10.
