复方青黛颗粒对大鼠溃疡性结肠炎模型的影响

目的 :观察复方青黛颗粒对大鼠溃疡性结肠炎模型的影响 ,为临床用药的有效性提供实验依据。方法 :采用异种异体结肠粘膜组织致敏法。将 SD大鼠随机分为空白对照组 ,模型对照组 ,复方青黛颗粒高、中、低剂量组 ,泻痢消胶囊组。连续给药 4周。空白对照组和模型对照组灌胃给予等量生理盐水 ;末次给药后 2 4 h,大鼠眼眶采血 ,测定乳酸脱氢酶和淀粉酶含量 ;处死大鼠 ,计数全结肠溃疡点和充血点 ;称结肠湿重 ,计算肠重指数。结果 :复方青黛颗粒 3个剂量组大鼠的溃疡数和充血指数显著减少或降低 ,与模型对照组比较差异有非常显著性意义 (P <0 .0 1) ,各剂量组大鼠的结肠重量和肠重指数 ,都有显著增加 ,中、高两剂量组与模型对照组比较差异有显著性意义(P <0 .0 5 ,<0 .0 1) ;血清乳酸脱氢酶含量较模型对照组有降低趋势 (P >0 .0 5 ) ,复方青黛颗粒组均有使模型动物降低的血清淀粉酶含量明显恢复的作用 ,与模型对照组比较差异有显著性意义 (P <0 .0 5或 <0 .0 1)。结论 :复方青黛颗粒对溃疡性结肠炎模型大鼠有显著的治疗作用     

复方青黛颗粒对溃疡性结肠炎模型大鼠结肠组织TGF-β1和VEGF表达影响

[目的]观察复方青黛颗粒对溃疡性结肠炎(UC)大鼠结肠组织转化生长因子-β1(TGF-β1)与血管内皮生长因子(VEGF)表达的影响,进一步探讨复方青黛颗粒治疗UC的相关机制。[方法]三硝基苯磺酸(TNBS)法制备大鼠UC模型,随机分为空白对照组、模型对照组、柳氮磺吡啶(SASP)组及复方青黛颗粒低、中、高剂量组。治疗结束后取大鼠结肠组织,用RT-PCR方法检测TGF-β1及VEGF mRNA表达。[结果]与空白对照组比较,模型对照组TGF-β1及VEGF基因表达明显增高(P0.05),复方青黛颗粒高剂量组TGF-β1基因表达较模型对照组显著下降(P0.05),VEGF基因表达较模型对照组显著升高(P0.05)。[结论]复方青黛颗粒对TNBS诱导的UC大鼠的治疗作用可能与下调TGF-β1过度表达及上调VEGF有关。     

青黛颗粒对溃疡性结肠炎大鼠结肠黏膜MUC2和iNOS基因表达的影响

目的:观察青黛颗粒对TNBS诱导的溃疡性结肠炎(UC)大鼠结肠黏膜黏蛋白(MUC2)及诱导型一氧化氮合成酶(iNOS)基因表达的影响,探讨其治疗UC的可能作用机制.方法:将54只SD实验大鼠随机分为正常对照组、模型对照组、阳性药物治疗组(SASP,500 mg/kg)、青黛颗粒600、900、1 200 mg/kg治疗组.造模后第3天开始灌胃给药,共给药10d.实验第14天,处死大鼠,剖取其病变结肠组织,比较各组大鼠的DAI积分和CMDI评分,用逆转录聚合酶链反应(RT-PCR)法检测MUC2及iNOS基因的表达.结果:较模型组青黛颗粒900、1200 mg/kg治疗组能显著降低实验大鼠DAI积分和CMDI评分,上调结肠组织中MUC2的基因表达(2.06±0.70 vs 1.24±0.47;2.34±0.86 vs 1.24±0.47.均P<0.01),且青黛颗粒1 200 mg/kg治疗组能下调iNOS的基因表达(0.35±0.12vs 0.62±0.31.P<0.05).结论:青黛颗粒可能通过上调结肠黏膜MUC2的基因表达并下调iNOS的基因表达而起到抗TNBS诱导的大鼠溃疡性结肠炎的作用.     

复方青黛颗粒对溃疡性结肠炎模型大鼠结肠MMP-1及TIMP-1表达的影响

[目的]观察复方青黛颗粒对溃疡性结肠炎（UC）模型大鼠结肠基质金属蛋白酶1（MMP-1）及其特异性组织抑制因子（TIMP-1）基因表达的影响,探讨其治疗UC的作用机制.[方法]用三硝基苯磺酸（TNBS）制备UC大鼠模型,将52只SD大鼠随机分为正常对照（空白）组、模型组、美沙拉嗪（5-ASA）组,复方青黛颗粒低、中、高剂量组,从造模后第3天开始分别每天灌胃给药1次至实验结束,第14天（灌胃10 d后）,用逆转录聚合酶链反应（RT-PCR）法检测MMP-1、TIMP-1基因表达的水平.[结果]模型组MMP-1、TIMP-1基因相对表达量均明显高于空白组（P＜0.05）;复方青黛颗粒中、高剂量组MMP-1相对表达量及中、高剂量组和5-ASA组TIMP-1相对表达量均明显低于模型组（均P＜ 0.05）.[结论]复方青黛颗粒能有效治疗TNBS诱导的UC模型大鼠,可能与复方青黛颗粒抑制MMP-1、TIMP-1基因表达有关.     

复方青黛颗粒联合路优泰治疗溃疡性结肠炎的疗效观察

[目的]观察复方青黛颗粒联合路优泰治疗溃疡性结肠炎(UC)的疗效。[方法]选取我科近3年病房收治的UC伴焦虑患者,将其分为复方青黛颗粒联合路优泰治疗组和复方青黛颗粒对照组,比较2组的疗效。[结果]治疗组和对照组疗效比较,差异具有统计学意义(P<0.05)。[结论]对于伴有焦虑的UC患者在治疗时给予抗焦虑药物具有一定的临床治疗意义。     

复方青黛颗粒对溃疡性结肠炎大鼠NF-κBP65、TNF-α表达的影响

目的:探讨复方青黛颗粒治疗溃疡性结肠炎(UC)大鼠的相关机制.方法:用三硝基苯磺酸(TNBS)法制备大鼠UC模型,分为空白对照组、模型对照组、柳氮磺吡啶(SASP)组、复方青黛颗粒低、中、高剂量组.造模后第3天开始灌胃给药,共给药10d,实验第14天,处死大鼠.取大鼠结肠组织及血清,用免疫组织化学SP法检测NF-κBP65蛋白表达,ELISA测定血清中肿瘤坏死因子α(TNF-α)的含量.结果:空白对照组与模型对照组比较,结肠组织中NF-κBP65蛋白表达及血清中TNF-α表达明显增高(0.276±0.0081vs0.138±0.003;67.657±3.580vs18.990±3.964,均P<0.05)复方青黛颗粒高剂量组与模型对照组相比,结肠组织中NF-κBP65蛋白表达及血清中TNF-α表达显著降低(0.217±0.007vs0.276±0.008;27.783±2.867vs67.657±3.580,均P<0.05).结论:复方青黛颗粒对TNBS诱导的UC大鼠的治疗作用可能与通过NF-κB信号传导通路调节TNF-α含量有关.     

青黛颗粒对溃疡性结肠炎实验大鼠血清白细胞介素6和10水平的影响

[目的]观察青黛颗粒对溃疡性结肠炎（UC）实验大鼠血清白细胞介素6（IL-6）和白细胞介素10（IL-10）水平的影响,探讨其治疗UC可能的作用机制。[方法]将52只SD大鼠采用三硝基苯磺酸（TNBS）法制备UC大鼠模型。造模结束后第2天取4只大鼠处死确定造模成功,余48只随机均分为6组：正常组,模型组,柳氮磺胺嘧啶（SASP）阳性药物治疗（SASP）组,青黛颗粒低、中、高剂量组,各8只。造模后第3天开始各组动物灌胃给药,连续给药10 d,实验第14天通过腹主动脉采血取血清,用ELISA法测定血清IL-6、IL-10水平。[结果]青黛颗粒低、中、高3个剂量组与模型组比较,血清IL-6水平显著降低,IL-10水平升高（P〈0.01）;青黛颗粒高剂量组血清IL-6I、L-10水平变化与SASP组比较,差异无统计学意义（P〉0.05）。[结论]青黛颗粒治疗实验性UC的作用机制可能与降低IL-6水平,升高IL-10水平有关。     

益生菌对实验性结肠炎大鼠Toll样受体4表达的影响

目的 观察两种益生菌(乳酸杆菌、双歧杆菌)对溃疡性结肠炎的大鼠结肠黏膜Toll样受体4(TLR4)及TNT-α的变化及影响.方法 采用三硝基苯磺酸/乙醇与免疫复合物联合诱导溃疡性结肠炎大鼠模型.观察和评估其结肠黏膜的大体和组织学变化.以免疫组化方法检测TIR4、TNF-α的表达,以蛋白免疫印迹法检测蛋白表达.结果 造模后结肠组织中可见大量炎性细胞浸润,杯状细胞减少,隐窝脓肿形成.与正常对照组相比,模型组结肠组织TLR4的表达显著增加(P＜0.01).TNF-α表达显著升高(P＜0.01);与益生菌组相比,模型组结肠组织TLR4的表达显著减少(P＜0.01),TNF-α表达显著减少(P＜0.01).其两种菌相比差异不大(P＞0.05).结论 从免疫机制的视角观察到益生菌可能通过抑制Toll样受体4表达从而减少炎性因子TNF-α产生.     

Toll样受体4的信号转导与溃疡性结肠炎的研究进展

溃疡性结肠炎(ulcerative colitis, UC)的发病机制尚不明确,目前研究表明,Toll样受体4(Toll-like receptor 4,TLR4)与UC的发病密切相关,TLR4通过髓样分化因子88 (myeloid differentiation factor 88, MyD88)依赖性和非依赖性等途径激活下游信号分子,引发肠黏膜释放TNF-α、IL-1、IL-6等炎症介质,最终导致UC的发生,同时对此信号转导通路的阻断研究为临床治疗UC带来了新的契机.     

清肠栓对葡聚糖硫酸钠诱导溃疡性结肠炎大鼠Toll样受体表达的影响

目的探讨清肠栓对葡聚糖硫酸钠诱导溃疡性结肠炎大鼠Toll样受体(TLRs)表达的影响。方法选取大鼠60只,根据随机数字表法分为对照组、模型组、清肠栓组,每组20只,然后对大鼠进行模型建造,造模时对照组给予蒸馏水,其余组均给予5%的葡聚糖硫酸钠。模型建立后清肠栓组给予清肠栓灌肠,比较大鼠疾病活动指数(DAI)、结肠指数以及TLR1、TLR2、TLR3以及TLR4表达。结果模型组DAI评分显著高于对照组,清肠栓组显著低于模型组(P0.05);清肠栓组结肠指数显著低于模型组(P0.05),与对照组比较无统计学意义(P0.05);模型组TLR2和TLR4表达显著高于对照组,清肠栓组TLR2和TLR4表达显著低于模型组(P0.05),各组TLR1和TLR3比较无统计学意义(P0.05)。结论清肠栓对葡聚糖硫酸钠诱导溃疡性结肠炎疗效显著,能显著降低大鼠TLR2和TLR4表达水平。     

Bacterial invasion into the colonic mucosa in ulcerative colitis

Abstract This study investigated interactions between mucosal lesions and bacterial invasion in ulcerative colitis using the acridine-orange staining method. In all 16 cases of ulcerative colitis, the mucosa was found to be invaded by small rods and cocci. In five of 10 controls, bacteria were seen only adhering to the mucosa and no bacteria were detected in the five remaining cases. It is suggested that the presence of bacteria in the colonic mucosa may be a factor responsible for the persistence or aggravation of ulcerative colitis.     

Genetic polymorphisms of CD14 and Toll-like receptor-2 (TLR2) in patients with ulcerative colitis

BACKGROUND AND AIM: Ulcerative colitis (UC) is a multifactorial disease resulting from a complex interaction of genetic and environmental factors. Identifying genetic variants that alter the innate immune response is highly relevant to understanding the pathogenesis of UC. The aim of this study was to investigate the association between CD14 and Toll-like receptor-2 (TLR2) genetic polymorphisms and chronic UC in Japanese patients. METHODS: The study population consisted of 102 chronic UC patients and 146 healthy control subjects. Polymorphisms in the promoter at C-260T of CD14 gene were investigated by PCR restriction fragment length polymorphism, and -196 to -174 del of TLR2 was investigated by allele-specific PCR. RESULTS: The frequencies of CD14 TT and T carrier were significantly higher in UC patients than in controls (TT: OR = 3.98, 95% CI 1.82-8.71, P = 0.0005; T carrier: OR = 2.98, 95% CI 1.47-6.01, P = 0.002). In addition, TT and T carrier were more closely associated with distal colitis phenotype (TT: OR = 7.78, 95% CI 2.14-28.28, P = 0.0007; T carrier: OR = 6.30, 95% CI 2.71-14.58, P = 0.005), onset after 20 years of age (TT: OR = 5.28, 95% CI 2.18-12.79; T carrier: OR = 3.79, 95% CI 1.67-8.59), chronic continuous type (TT: OR = 4.26, 95% CI 1.56-11.64; T carrier: OR = 3.09, 95% CI 1.33-7.82), and fewer than two hospitalizations (TT: OR = 4.44, 95% CI 1.81-10.89; T carrier: OR = 3.26, 95% CI 1.43-7.27). There was no significant difference in TLR2 -196 to -174 del/del and del/ins carrier frequencies between UC patients and healthy controls. However, these frequencies were significantly higher in steroid-dependant patients than in controls (del/del: OR = 6.08, 95% CI 1.41-26.21; del carrier: OR = 3.00, 95% CI 1.13-7.98). CONCLUSION: The results suggest that existence of a mutation in the CD14 gene is associated with an increased susceptibility to developing UC, especially chronic continuous distal colitis phenotypes that develop after 20 years of age. Furthermore, polymorphism of TLR2 may be related to an increased risk of intensive types such as steroid-dependent patients.     

Telomere shortening in the colonic mucosa of patients with ulcerative colitis

Telomere length in human somatic cells gradually decreases with the number of cell divisions and is regarded as a marker of somatic cell turnover. Mucosal cells of the affected colon show rapid turnover in individuals with active ulcerative colitis (UC). Telomere length was determined by Southern blot analysis of terminal restriction fragments (TRFs) from the colonic mucosa of 17 patients with UC in remission, two of whom showed dysplasia, and 17 control subjects without colitis. For each individual, mean TRF length was compared between rectal mucosa and unaffected cecal mucosa. The mean TRF length of the rectal mucosa was significantly less than that of cecal mucosa in UC patients (7.87 ± 0.36 kb versus 8.77 ± 0.21 kb; P = 0.0015, Wilcoxon signed rank test), whereas no significant difference was detected in the control subjects. The extent of telomere shortening was 10.6 ± 3.35% in UC patients, compared with 0.8 ± 0.64% in noncolitis controls (P = 0.0024, Mann-Whitney U-test). Four UC patients, two of whom had dysplasia, showed telomere shortening of more than 20% in the rectal mucosa. These observations suggest that telomere shortening in the colonic mucosa of individuals with UC may represent the history of mucosal inflammation during disease of long duration, and that it may contribute to aneuploidy in UC. (Received May 6, 1997; accepted Sept. 26, 1997)     

Differential expression of CCR5 and CRTH2 on infiltrated cells in colonic mucosa of patients with ulcerative colitis

BACKGROUND AND AIM: The pathogenesis of ulcerative colitis (UC) is unclear, but abnormal infiltration of T lymphocytes in the colonic mucosa has been implicated in the mucosal tissue damage. The abnormal cytokine production because of a T helper (h)1/Th2 imbalance may play an important role in continuing inflammation in the colonic mucosa. In the present study, the expression of chemokine receptor 5 (CCR5) as a Th1 marker and a chemoattractant receptor-homologs molecule expressed on Th2 cells (CRTH2) were investigated in order to analyze impaired Th1/Th2 responses in the colonic mucosa of UC patients. METHODS: Tissue samples were obtained by colonic biopsies from patients with UC or colonic polyps, with informed consent. Immunohistochemical analysis was performed on periodate, lysine-paraformaldehyde-fixed serial cryostat sections using the labeled streptavidin biotin method. Monoclonal antibodies against CD4, CCR5 or CRTH2 were used as primary antibodies. The number of cells expressing CD4, CCR5 or CRTH2 per unit area was calculated by using an image analyzer. RESULTS: In the patients with UC, the numbers of CD4- and CCR5-positive cells were significantly increased in inflamed mucosa, and appeared to be correlated with the disease activity. The infiltration of CRTH2-positive cells was predominantly observed in the mildly inflamed or the margin of inflamed mucosa of UC patients. CONCLUSION: There is a possibility that Th1 responses significantly occur in colonic mucosa with severe inflammation, while Th2 responses mainly occur with mild inflammation in UC patients. The Th1/Th2 imbalance in colonic mucosa may be related to the disease progression of UC.     

Tissue-type plasminogen activator of colonic mucosa in ulcerative colitis

Microvascular endothelial alterations are thought to be a crucial step for development of hemorrhagic changes in various pathological states. In this study, we determined the activity and amount of tissue-type plasminogen activator (t-PA) in the biopsy specimens from sigmoid colon of patients with ulcerative colitis to evaluate endothelial alterations and vascular changes of permeability. The results of this investigation revealed that mucosal amount of t-PA in the active stage of ulcerative colitis was two- to threefold higher than in healthy controls, while t-PA levels in plasma samples showed no remarkable differences among the groups. Increased t-PA activity appeared to correlate well to the degree of inflammation of colonic mucosa, and t-PA amount was still increased in the inactive stage. The present study indicates that t-PA determination in colonic biopsy specimens may be useful for the evaluation of clinical activity of ulcerative colitis in terms of the mucosal microvascular endothelial changes.This work was supported by the grant from the Japanese Ministry of Education No. 63440033 and by the grant from Keio University, School of Medicine.     

TLR2在溃疡性结肠炎患者结肠黏膜中的表达

目的 检测溃疡性结肠炎(UC)患者结肠肠黏膜TLR2蛋白及TLR2 mRNA的表达及其与UC临床活动度、内镜分级的关系.方法 取47例UC患者结肠黏膜标本根据内镜及临床活动度进行分级,同时收集正常对照者结肠黏膜标本13例.用Western Blot技术和半定量反转录聚合酶链反应方法(RT-PCR)检测结肠黏膜中TLR2蛋白及TLR2 mRNA的表达.结果 UC患者结肠黏膜TLR2蛋白及TLR2mRNA的表达量高于正常对照者,且随着内镜下及临床病变分级的加重逐渐增加.结论 UC患者肠黏膜中TLR2及TLR2 mRNA的表达增加,且其表达均随临床及内镜下病变程度加重,表达逐渐增加,在一定程度上可以反映疾病的活动度.