CAJ | 학술논문

目的探讨姜黄素在三硝基苯磺酸(TNBS)诱导的大鼠结肠纤维化中的抗纤维化作用和机制.方法 SD大鼠40只随机分组,模型组(10只)、治疗组(10只)和对照组(10只)分别于第1、8、15、22和29天予TNBS 10 mg、15 mg、20 mg、25 mg和30 mg灌肠,另取10只大鼠给予50%乙醇灌肠,作为阴性对照(正常组).从实验周期第1天起,治疗组大鼠每日予姜黄素30 mg/kg腹腔注射,对照组每日予0.9%NaCl腹腔注射,模型组和正常组不予处理.采用HE染色及Masson胶原三色染色观察大鼠结肠组织损伤和纤维化变化,采用ELISA法检测结肠黏膜中Th1/Th2型细胞因子IL-2、TNF-α、IL-4、IL-17的含量,采用荧光定量PCR法检测结肠黏膜中肠纤维化相关细胞因子如转化生长因子(TGF)-β1、结缔组织生长因子(CTGF)、Smad3、胶原Ⅰ、ⅢmRNA的表达.结果模型组与对照组大鼠结肠组织大体损伤评分[(6.14±1.07)分,(6.17±1.47)分]及组织损伤评分[(8.42±1.40)分,(8.17±1.47)分]、胶原面积比(36.59%±4.07%,37.18%±4.05%)较正常组[分别为(2.13±0.64)分,(2.25±1.28)分和25.43%±5.39%]明显升高(均P＜0.05).模型组与对照组大鼠黏膜组织中IL-2[(378.25±29.90)ng/L,(410.06±64.74)ng/L]、TNF-α[(87.11±23.85)ng/L,(100.41±12.59)ng/L)]、IL-17[(47.80±5.62)ng/L,(41.45±2.12)ng/L]含量及TGF-β1(4.71%±2.71%,4.12%±3.01%)、CTGF(10.33%±6.99%,11.46%±4.72%)、Smad3(9.35%±7.32%,10.11%±3.80%)、胶原Ⅰ(1.52%±1.11%,1.57%±1.35%)、胶原Ⅲ(3.04%±1.33%,3.03%±3.53%)mRNA表达量较正常组[分别为(179.74±20.73)ng/L,(35.47±7.13)ng/L,(14.48±7.52)ng/L和0.90%±1.13%,0.53%±0.47%,0.62%±0.44%,0.16%±0.09%,0.18%±0.10%]均明显升高(均P＜0.05).而治疗组大鼠结肠组织大体损伤评分[(4.00±1.07)分]及组织损伤评分[(5.13±1.46)分]、胶原面积比(30.01%±7.56%),IL-2[(223.91±28.04)ng/L]、TNF-α[(44.19±4.77)ng/L]、IL-17[(14.89±4.31)ng/L]含量和TGF-β1(0.85%±0.76%)、CTGF(1.56%±1.13%)、Smad3(3.62%±3.03%)、胶原Ⅰ(0.40%±0.31%)、胶原Ⅲ(0.60%±1.02%)mRNA表达量较模型组及对照组明显降低(P＜0.05),与正常组无明显差异(P＞0.05).结论姜黄素可通过降低细胞因子的过度表达,减轻大鼠结肠炎症,从而抑制过度"损伤-修复"所致的组织纤维化. 目的探討薑黃素在三硝基苯磺痠(TNBS)誘導的大鼠結腸纖維化中的抗纖維化作用和機製.方法 SD大鼠40隻隨機分組,模型組(10隻)、治療組(10隻)和對照組(10隻)分彆于第1、8、15、22和29天予TNBS 10 mg、15 mg、20 mg、25 mg和30 mg灌腸,另取10隻大鼠給予50%乙醇灌腸,作為陰性對照(正常組).從實驗週期第1天起,治療組大鼠每日予薑黃素30 mg/kg腹腔註射,對照組每日予0.9%NaCl腹腔註射,模型組和正常組不予處理.採用HE染色及Masson膠原三色染色觀察大鼠結腸組織損傷和纖維化變化,採用ELISA法檢測結腸黏膜中Th1/Th2型細胞因子IL-2、TNF-α、IL-4、IL-17的含量,採用熒光定量PCR法檢測結腸黏膜中腸纖維化相關細胞因子如轉化生長因子(TGF)-β1、結締組織生長因子(CTGF)、Smad3、膠原Ⅰ、ⅢmRNA的錶達.結果模型組與對照組大鼠結腸組織大體損傷評分[(6.14±1.07)分,(6.17±1.47)分]及組織損傷評分[(8.42±1.40)分,(8.17±1.47)分]、膠原麵積比(36.59%±4.07%,37.18%±4.05%)較正常組[分彆為(2.13±0.64)分,(2.25±1.28)分和25.43%±5.39%]明顯升高(均P＜0.05).模型組與對照組大鼠黏膜組織中IL-2[(378.25±29.90)ng/L,(410.06±64.74)ng/L]、TNF-α[(87.11±23.85)ng/L,(100.41±12.59)ng/L)]、IL-17[(47.80±5.62)ng/L,(41.45±2.12)ng/L]含量及TGF-β1(4.71%±2.71%,4.12%±3.01%)、CTGF(10.33%±6.99%,11.46%±4.72%)、Smad3(9.35%±7.32%,10.11%±3.80%)、膠原Ⅰ(1.52%±1.11%,1.57%±1.35%)、膠原Ⅲ(3.04%±1.33%,3.03%±3.53%)mRNA錶達量較正常組[分彆為(179.74±20.73)ng/L,(35.47±7.13)ng/L,(14.48±7.52)ng/L和0.90%±1.13%,0.53%±0.47%,0.62%±0.44%,0.16%±0.09%,0.18%±0.10%]均明顯升高(均P＜0.05).而治療組大鼠結腸組織大體損傷評分[(4.00±1.07)分]及組織損傷評分[(5.13±1.46)分]、膠原麵積比(30.01%±7.56%),IL-2[(223.91±28.04)ng/L]、TNF-α[(44.19±4.77)ng/L]、IL-17[(14.89±4.31)ng/L]含量和TGF-β1(0.85%±0.76%)、CTGF(1.56%±1.13%)、Smad3(3.62%±3.03%)、膠原Ⅰ(0.40%±0.31%)、膠原Ⅲ(0.60%±1.02%)mRNA錶達量較模型組及對照組明顯降低(P＜0.05),與正常組無明顯差異(P＞0.05).結論薑黃素可通過降低細胞因子的過度錶達,減輕大鼠結腸炎癥,從而抑製過度"損傷-脩複"所緻的組織纖維化.
목적 탐토강황소재삼초기분광산(TNBS)유도적대서결장섬유화중적항섬유화작용화궤제.방법 SD대서40지수궤분조,모형조(10지)、치료조(10지)화대조조(10지)분별우제1、8、15、22화29천여TNBS 10 mg、15 mg、20 mg、25 mg화30 mg관장,령취10지대서급여50%을순관장,작위음성대조(정상조).종실험주기제1천기,치료조대서매일여강황소30 mg/kg복강주사,대조조매일여0.9%NaCl복강주사,모형조화정상조불여처리.채용HE염색급Masson효원삼색염색관찰대서결장조직손상화섬유화변화,채용ELISA법검측결장점막중Th1/Th2형세포인자IL-2、TNF-α、IL-4、IL-17적함량,채용형광정량PCR법검측결장점막중장섬유화상관세포인자여전화생장인자(TGF)-β1、결체조직생장인자(CTGF)、Smad3、효원Ⅰ、ⅢmRNA적표체.결과 모형조여대조조대서결장조직대체손상평분[(6.14±1.07)분,(6.17±1.47)분]급조직손상평분[(8.42±1.40)분,(8.17±1.47)분]、효원면적비(36.59%±4.07%,37.18%±4.05%)교정상조[분별위(2.13±0.64)분,(2.25±1.28)분화25.43%±5.39%]명현승고(균P＜0.05).모형조여대조조대서점막조직중IL-2[(378.25±29.90)ng/L,(410.06±64.74)ng/L]、TNF-α[(87.11±23.85)ng/L,(100.41±12.59)ng/L)]、IL-17[(47.80±5.62)ng/L,(41.45±2.12)ng/L]함량급TGF-β1(4.71%±2.71%,4.12%±3.01%)、CTGF(10.33%±6.99%,11.46%±4.72%)、Smad3(9.35%±7.32%,10.11%±3.80%)、효원Ⅰ(1.52%±1.11%,1.57%±1.35%)、효원Ⅲ(3.04%±1.33%,3.03%±3.53%)mRNA표체량교정상조[분별위(179.74±20.73)ng/L,(35.47±7.13)ng/L,(14.48±7.52)ng/L화0.90%±1.13%,0.53%±0.47%,0.62%±0.44%,0.16%±0.09%,0.18%±0.10%]균명현승고(균P＜0.05).이치료조대서결장조직대체손상평분[(4.00±1.07)분]급조직손상평분[(5.13±1.46)분]、효원면적비(30.01%±7.56%),IL-2[(223.91±28.04)ng/L]、TNF-α[(44.19±4.77)ng/L]、IL-17[(14.89±4.31)ng/L]함량화TGF-β1(0.85%±0.76%)、CTGF(1.56%±1.13%)、Smad3(3.62%±3.03%)、효원Ⅰ(0.40%±0.31%)、효원Ⅲ(0.60%±1.02%)mRNA표체량교모형조급대조조명현강저(P＜0.05),여정상조무명현차이(P＞0.05).결론 강황소가통과강저세포인자적과도표체,감경대서결장염증,종이억제과도"손상-수복"소치적조직섬유화.
Objective To investigate the anti-fibrotic effects of curcumin in trinitrobenzene sulphonic acid (TNBS) induced intestinal fibrosis in rats and its mechanism. Methods Forty SD rats were randomly divided into model group, treatment group, control group and normal group with 10each. Except the normal group, the other three groups were given 10, 15, 20, 25 and 30 mg of TNBS enema on the 1st, 8 th, 15th, 22nd and 29th days,respectively. The rats in treatment group were intraperitonealy injected with 30 mg/kg of curcumin daily. Control group was injected with 0. 9%NaCl solution and normal group received an equal volume of 50% ethanol enema without any treatment. The damage and fibrosis of colon were detected with HE staining and Masson collagen staining, respectively. The contents of interleukin (IL) -2, tumor necrosis factor (TNF) -α, IL-4 and IL-17 in colon were measured by enzyme-link immunosorbent analysis (ELISA). The expressions of intestinal fibrosis related cytokines such as transforming growth factor (TGF) -β1, connective tissue growth factor (CTGF), Smad3, collagen Ⅰ and collagen Ⅲ mRNA were determined by FQ-PCR.Results The macroscopic and micrpscopic colonic damage scores and collagen area were significantly higher in model group (6.14 ± 1.07, 8. 42 ± 1.40 and 36. 59% ± 4.07%, respectively) and control group (6.17 ± 1.47, 8. 17 ±1.47 and 37.18 %±4.05 %, respectively) than those in normal group (2.13±0.64, 2.25±1.28 and 25.43%±5.39% ,respectively)(P＜0.05). Contents of IL2, TNF-α, IL-17, as well as expressions of intestinal fibrosis related cytokines including TGF-β1, CTGF,Smad3, collagen Ⅰ and Ⅲ mRNA were also higher in model group [(378. 25±29. 90) ng/L,(87.11±23.85) ng/L, (47.80±5.62) ng/L, 4.71%±2.71%,10.33%±6.99%,9.35%±7.32%,1.52% ± 1.11% and 3.04% ±1.33%, respectively] and control group [(410. 06 ± 64.74) ng/L,(100.41±12.59) ng/L, (41.45±2. 12) ng/L, 4. 12%±3.01%,11.46%±4.72%,10. 11%±3.80%,1. 57% ± 1. 35% and 3. 03% ± 3. 53%, respectively] in comparision with normal group [(179.74±20. 73) ng/L, (35. 47±7. 13) ng/L, (14. 48±7. 52) ng/L and 0. 90%± 1. 13%,0.53%±0.47%, 0. 62%±0. 44%, 0. 16%±0. 09% and 0. 18%±0. 10%, respectively] (P＜0.05). While in treatment group, the macroscopic (4.00 ± 1.07 ) and micrpscopic (5. 13 ± 1.46)colonic damage scores, collagen area (30.01%±7.56%), contents of IL-2 [(223.91±28.04) ng/L],TNF-α [(44.19±4. 77) ng/L] and IL-17 [(14.89±4. 31) ng/L], expressions of TGF-β1 (0.85%±0.76%), CTGF (1.56%±1.13%), Smad3 (3.62%±3.03%), collagen Ⅰ (0.40%±0.31%) and Ⅲ (0.60 % ± 1.02 % ) mRNA were much lower than those in model group and control group (P＜0.05 ), but similar to those in normal group (P＞ 0.05 ). Conclusions Curcumin can inhibit intestinal fibrosis caused by excessive "wound-healing" reaction via reducing the overexpression of cytokines in colonic mucosa and attenuating the inflammation of colon.