肠安康胶囊对大鼠溃疡性结肠炎结肠组织细胞凋亡的影响

目的探讨肠安康胶囊对大鼠溃疡性结肠炎（UC）结肠组织细胞凋亡的影响。方法三硝基苯磺酸乙醇溶液大鼠灌肠建立UC模型，将大鼠随机分为正常组、模型组、柳氮磺胺吡啶（SASP）组和肠安康大、中、小剂量组，原位末端标记染色法检测结肠组织细胞凋亡，免疫组织化学法检测Fas／Fas-L、Bas／Bcl-2蛋白，RT—PCR法检测Caspase-3和c-Myc基因mRNA。结果与正常组比较，模型组结肠组织凋亡细胞明显增多（P〈0．01），肠安康胶囊治疗后，捅亡细胞数目明显减少（P〈0．01），并明显降低了凋亡相关调控基因Caspase-3、c-MycmRNA及Fas／Fas-L、Bas／Bcl-2蛋白的表达（P〈0．01）。结论肠安康胶囊通过改善相关细胞凋亡蛋白及基因的表达对实验性UC有一定的治疗作用。     

理肠四方对溃疡性结肠炎大鼠结肠组织TNF-αmRNA表达的影响

目的:观察理肠四方对溃疡性结肠炎(UC)大鼠结肠组织细胞因子TNF-αmRNA的表达的影响,比较四方疗效大小,并分析其作用机制.方法:应用2,4-二硝基氯苯(DNCB)免疫加醋酸局部灌肠法建立UC大鼠模型,将98只健康SD大鼠(雌雄各半),按雌雄随机分7组,分别为乌梅丸组、白头翁汤组、参苓白术散组、痛泻要方组、柳氮磺胺吡啶(SASP)组、模型组和正常组,分别观察治疗后大鼠结肠黏膜TNF-omRNA的表达变化,并进行统计学比较.结果:造模后细胞因子TNF-αmRNA的表达升高(模型组与正常组平均吸光度比较P＜0.01,t=4.128 vs正常组),理肠四方各治疗组、SASP组与模型组比较有显著意义(P＜0.01,q=12.37vs乌梅丸组,q=9.52 vs白头翁汤组,q=8.79vs参苓白术散组,q=8.54 vs痛泻要方组,q=8.92 vsSASP组),但除乌梅丸外,理肠四方之间比较无明显差异.结论:经两两比较后,乌梅丸疗效最好,白头翁汤、参苓白术散、痛泻要方与SASP组疗效相当.表明理肠四方可下调促炎细胞因子TNF-α,从而使溃疡性结肠炎大鼠免疫功能恢复正常,达到治疗的目的.     

肠安康胶囊对大鼠溃疡性结肠炎结肠组织细胞凋亡及细胞因子表达的影响

目的探讨肠安康胶囊对实验性溃疡性结肠炎结肠组织细胞凋亡及炎性细胞因子表达的影响。方法采用三硝基苯磺酸乙醇溶液给大鼠灌肠建立溃疡性结肠炎模型,将大鼠随机分为对照组、模型组、柳氮磺胺吡啶（sAsP）组和肠安康大、中、小剂量组,采用TUNEL染色法观察各组结肠组织细胞凋亡的情况,并采用ELISA法测定结肠组织的TNF-α、IFN-γ、IL-2、IL-6、IL-8。结果与对照组比较,模型组结肠组织凋亡细胞明显增多（P〈0.01）,且细胞凋亡的程度与TNF-α、IFN-γ、IL-2、IL-6、IL-8的水平分别呈正相关性（r分别为0.703、0.906、0.920、0.756、0.765,P〈0.01）。与模型组比较,各药物处理组结肠组织中凋亡细胞数目明显减少（P〈0.01）,TNF-α、IFN-γ、IL-2、IL-6、IL-8水平显著上升（P〈0.01）。结论细胞凋亡是溃疡性结肠炎的发病机制之一,凋亡程度与相关炎症因子水平密切相关,肠安康胶囊通过改善相关炎性细胞因子表达的水平,对实验性溃疡性结肠炎有一定的治疗作用。     

理肠四方治疗大鼠溃疡性结肠炎疗效观察

目的:观察理肠四方对溃疡性结肠炎(UC)大鼠不同的组织形态学变化,比较四方疗效大小,并分析其中医病机.方法:应用2,4-二硝基氯苯免疫加醋酸局部灌肠法建立UC大鼠模型,将98只健康SD大鼠(雌雄各半),按雌雄随机分为7组[乌梅丸组、白头翁汤组、参苓白术散组、痛泻要方组、柳氮磺胺吡啶(SASP)组、模型组和正常组],每组14只,分别观察治疗后大鼠一般情况变化、结肠组织损伤评分、结肠粘膜病理及超微病理变化.结果:经理肠四方治疗后,其组织学呈不同的病理变化.结论:乌梅丸疗效最好,白头翁汤、参苓白术散、痛泻要方与SASP疗效相当.     

鱼腥草治疗溃疡性结肠炎大鼠对结肠压力的影响

溃疡性结肠炎患者（ＵＣ）经常有腹痛,腹泻等临床表现,这些症状与结肠动力紊乱密切相关［１,２］,但目前尚缺少一种理想的检测结肠动力的方法,如常用的大鼠离体结肠压力测定,不仅需要麻醉,手术,开腹,而且离体肠段保存条件很难与在体状态一致,影响了检测结果的准...     

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.     

细胞凋亡与溃疡性结肠

0 引言 溃疡性结肠炎(ulcerative colitis,UC)是原因不明的慢性肠道炎症性疾病,目前人们认为他的发病机制是多因素相互作用的结果,包括环境因素、遗传因素和免疫因素等,即感染、饮食等环境因素作用于遗传易感的人群,使肠免疫反应过度亢进,导致肠黏膜损伤而发病。但UC的病因仍然不清楚。近年来,UC与细胞凋亡的关系受到了人们的关注,一些结果显示细胞凋亡可能在UC的发病中起重要的作用,下面我们从细胞凋亡的角度来探讨一下UC的发病机制。     

结肠康泰对溃疡性结肠炎大鼠结肠黏膜细胞增殖及周期的影响

[目的]探讨结肠康泰对溃疡性结肠炎（UC）大鼠模型结肠黏膜细胞增殖及周期的影响。[方法]雌性SD大鼠60只，随机分为正常对照、模型、柳氮磺胺吡啶（SASP）及结肠康泰组，以5％2、4、6，三硝基苯磺酸（TNBs）诱导大鼠形成结肠炎症，免疫组化检测增殖细胞核抗原（PCNA）的表达及流式细胞仪分析细胞周期。[结果]SASP及结肠康泰组大鼠增殖细胞指数（LI）明显增高，G0／1期细胞明显减少而G2／M期细胞增加，与模型组比较差异均有统计学意义（均P〈0．05）。[结论]结肠康泰能增强UC大鼠结肠黏膜细胞的增殖能力。     

苦参碱对溃疡性结肠炎患者T细胞亚群的影响

[目的]探讨苦参碱注射液对溃疡性结肠炎（UC）患者外周血T细胞免疫的影响.[方法]采用免疫组化（SAP）法检测30例UC患者用苦参碱注射液治疗1月前、后和10例健康志愿者外周血T细胞亚群.[结果]30例UC患者中治愈7例,好转12例,无变化11例;UC患者CD8^＋Ts细胞明显减少;用苦参碱治疗后UC患者外周血CD3^＋、CD8^＋Ts细胞较治疗前明显增高,有效组CD8^＋Ts细胞显著高于无效组（P＜0.05）.[结论]UC患者抑制性T细胞减低,免疫亢进;用苦参碱注射液治疗有较好疗效,外周血T细胞,尤其CD8^＋Ts细胞水平明显增高.     

Autoimmunity in ulcerative colitis: humoral and cellular immune response bytropomyosin in ulcerative colitis

AIM Autoimmunity has been emphasized in the pathogenesis of ulcerative colitis (UC). We reported thattropomyosin (TM) or TM related protein is a putative autoantigen in UC. In human fibroblast, at least 8isoforms of TM have been identified with molecular weight range from 30kD to 40kD, depending upon theisoforms, and human TM isoforms (hTM5) has been found the main isoform in human intestinal epithelialcells. In this study, hTM5 was used as a putative auto-antigen for the humoral and T cell immune responses inpatients with UC, Crohn＇s disease (CD) and healthy subjects (HS) as controls.METHODS Anti-bTM antibody was examined by enzyme-linked immunosorbent assay using human sera(UC 59, CD 28, HS 26) against hTM isoforms. The IFN-γ production by peripheral blood T cells followingstimulation by recombinant hTM5 was analyzed by ELISPOT assay.RESULTS Anti-hTM5 antibody (IgG1) was detected in 15/59 (25.4％) patients with UC, 3/28 (10.γ％)with CD, and 3/26 (11.5％) of HS. The OD value in UC was significantly higher than in CD and HS groups(P ＜ 0.05; P ＜ 0.01 respectively). Western blot analysis demonstrated immunoreactivity against hTM5 inseveral UC sera. ELISPOT assay demonstrated that IFN-γ production is significantly higher in UC (7/18),39.0％), compared with CD (0/8, 0％) and HS (0/7, 0％), (P＜0.05).CONCLUSION A significantly higher immune response to hTM5 was present in UC compared to CD andHS. Further studies of the hTM5/peptides may provide immuno-biochemical mechanism of autoimmuneprocess in UC.     

溃疡性结肠炎多药耐药基因的表达及其与疾病行为间的关系

目的 探讨溃疡性结肠炎多药耐药基因（MDR1）产物P-gp和多药耐药相关蛋白（MRP）的表达及其与疾病行为间的关系.方法 应用多药耐药基因（MDR1）产物P-gp和多药耐药相关蛋白（MRP）单克隆抗体,对溃疡性结肠炎患者内镜活检组织进行免疫组化检测.结果 多药耐药基因（MDR1）产物P-gp和多药耐药相关蛋白（MRP）在溃疡性结肠炎病程12个月、18个月时表达率分别为40.5%、45.9%、48.6%和51.4%,分别与患病初期相比,差异有统计学意义（P〈0.05）;P-gp和MRP在溃疡性结肠炎活动期和缓解期表达率分别为36.4%、18.6%和46.1%、25.4%,两期分别相比差异有统计学意义（P〈0.05）;P-gp和MRP活动期轻度、中度和重度之间差异无统计学意义（P〉0.05）.结论 通过内镜活检组织P-gp和MRP检测可了解溃疡性结肠炎耐药情况,为临床治疗提供依据,方法安全可靠、简便可行.P-gp和MRP表达在活动期严重程度间无差异,不宜作为严重程度是否耐药判断指标.     

溃疡性结肠炎患者外周血中性粒细胞凋亡与粘附分子水平的关系及意义

目的　探讨溃疡性结肠炎 (U C)患者外周血中性粒细胞 (PMN)凋亡机制。方法　采用流式细胞术检测 32例 UC患者外周血 PMN凋亡 ,EL ISA法检测 P-选择素 (P- sel)和细胞间粘附分子 - 1(ICAM- 1)的水平。结果　活动期 UC患者 PMN凋亡率明显低于对照组和缓解期 UC患者 (P<0 .0 1)。不同病情活动期 U C患者 PMN凋亡有显著性差异 (P<0 .0 1)。活动期 UC患者外周血中 P- sel和 ICAM- 1水平均高于对照组和缓解期 U C患者(P<0 .0 1或 (P<0 .0 5 ) ,且与 PMN凋亡呈负相关 (r值分别为 - 0 .72 38和 - 0 .5 2 13,P均 <0 .0 1) ,与病情呈正相关。结论　各种免疫细胞粘附分子表达上调可能是导致 UC患者 PMN凋亡延迟的重要机制     

Altered expression of innate immunity genes in different intestinal sites of children with ulcerative colitis

Background

Innate immunity has been very rarely investigated in ulcerative colitis and never in paediatrics. The present study was aimed at describing expression of innate immunity genes (NOD2, RIP2, α-defensins HD5 and HD6) in inflamed colon and in ileum of children with ulcerative colitis. Expression of TNFα and IL-1β was also analyzed.

Methods

15 children with ulcerative colitis (9 pancolitis, 6 left-sided colitis) and 10 control children were enrolled. mRNA and protein expressions were detected by real time PCR and western blot assays.

Results

NOD2, RIP2, IL-1β, TNFα expression levels were significantly increased in colonic mucosa of patients compared to controls (p < 0.01). These genes were also upregulated (p < 0.01) in the ileum of both pancolitis and left-sided colitis children. HD5 and HD6 were significantly upregulated (p < 0.01) in the inflamed colon of patients as well as in the ileum of those with pancolitis.

Conclusions

An increased mucosal expression of innate immunity genes was found in the inflamed colon of children with ulcerative colitis, outlining the role of the innate immune response in disease pathogenesis. Involvement of the ileum in ulcerative colitis suggests that an immune activation can also be established in intestinal sites classically uninvolved by the inflammation, carrying implications for the treatment and course of the disease.     

Effect of traditional Chinese medicinal enemas on ulcerative colitis of rats

AIM: To investigate the effects of traditional Chine semedicinal enema (TCME) on inflammatory and immune response of colonic mucosa of rats with ulcerative colitis(UC), and to observe the pathogenic mechanism.METHODS: Thirty UC rats, induced by intestinal enema together with 2.4-dinitrochlorobenzene (DNCB) and acetic acid, were randomly divided into 3 groups, i.e., GⅠ, GⅡ and GⅢ. Groups GⅠ and GⅡ were administered with TCME and salazosulfapyridine enema (SASPE), respectively. Group GⅢ was clystered with only normal saline (NSE), served as control. Group GIV was taken from normal rats as reference,once daily, from the 7th day after the establishment of UC for total 28 d. Interleukin-6 (IL-6) in the colonic mucosa was assayed by ^3H-TdR incorporation assay. Colonic mucosal lymphocyte subpopulation adhesive molecules, CD4^+CD11a^+,CD4^+CD18^+, CDs^+CD11a^+, CDs^+CD18^+ (LSAM), tumor necrosis factor (TNF)-α, and interferon-γ (IFN-γ), were detected by enzyme linked immunosorbent assay (ELISA). Moreover, the expression of TNF-α mRNA and IFN-γ mRNA in colonicmuc osa were detected by polymerase chain reaction (RT-PCR).RESULTS: Before therapies, in model groups, GⅠ, GⅡ and GⅢ,levels of IL-6, TNF-α, IFN-γ, CDs^+CD11a^+ and CD8^+CD18^+ were significantly different (38.29+2.61 U/mL, 16.54+1.23 ng/L,8.61+0.89 ng/L, 13.51+2.31% and 12.22+1.13% ,respectively) compared to those in GIV group (31.56+2.47 U/mL, 12.81+1.38 ng/L, 5.28+0.56 ng/L, 16.68+1.41% and 16.79+1.11%, respectively). After therapeutic enemas,in GI group, the contents of IL-6 (32.48&#177;2.53 U/m), TNF-α(13.42&#177;1.57 ng/L) and IFN-γ (5.87+0.84 ng/L) were reduced; then, the contents of CD8^+CD11a^+ (16.01+1.05 %)and CD8^+CD18^+ (16.28&#177;0.19%) were raised. There was no significant difference between groups GⅠ and GIV, but the difference between groups GⅠ and GⅡ was quite obvious (P&lt;0.05). The expressions of TNF-α mRNA and IFN-γ mRNAin group GⅢ were much higher than those of group GIV,but those in group GI were significantly suppressed by TCME therapy.CONCLUSION: Ulcerative colitis is related to colonic regional mucosal inflammatory factors and immune imbalance. TCME can effectively inhibit regional mucosal inflammatory factors and improve their disorder of immunity.     

HLA-DR antigen expression on colonic epithelium of ulcerative colitis in comparison with infectious colitis and ischemic colitis

We examined HLA-DR antigen expression on endoscopically biopsied colonic epithelium of ulcerative colitis (UC), infectious colitis and ischemic colitis. Since this monoclonal antibody (LN-3 ICN Immunobiological, USA) is available for usual formalin fixated materials, if the fixation is limited within 36 hours. 886 samples from 55 UC cases, 91 samples from 19 infectious colitis cases, 63 samples from 15 ischemic colitis cases and 63 samples from normal cases were enough statistically, compared to DR antigen expression. UC expressed clearly statistical high positive DR staining rate than infections colitis and ischemic colitis. Further, samples from UC and infectious colitis were compared in the histopathologically each with the same grade of inflammation, UC expressed higher positive rates of DR antigen than infectious colitis, and both UC and infectious colitis showed increased positive rates of DR antigen with advance of histopathological grades of inflammation.     

电针对大鼠脑缺血再灌注损伤后细胞凋亡相关基因Bcl-2、Bax表达的影响

目的探讨电针对脑缺血再灌注大鼠皮质细胞凋亡相关蛋白Bcl-2、Bax及其mRNA表达的影响。方法雄性清洁级SD大鼠72只,采用随机数字表法分为假手术组、模型组和电针组各24只。用改良Longa线栓法制作大鼠右侧大脑中动脉脑缺血模型,缺血2 h,于再灌注开始后电针组针刺"百会"和"大椎"穴。各组于缺血再灌注24 h后行神经行为学评分和脑含水量测定,免疫组化染色法检测缺损侧皮层组织Bcl-2、Bax的蛋白表达,qRT-PCR检测Bcl-2 mRNA、Bax mRNA表达的变化。结果模型组、电针组神经行为学评分均低于假手术组(P<0.01),与模型组比较,电针组的神经行为学评分升高(P<0.05)。与假手术组比较,模型组脑含水量明显增高(P<0.01),电针组的差异无统计学意义(P>0.05),较之模型组,电针组的脑含水量显蓍降低(P<0.01)。与假手术组比较,模型组、电针组Bcl-2蛋白及mRNA的表达均显著增多(P<0.05或P<0.01),电针组又高于模型组(P<0.05);较之假手术组,模型组Bax蛋白及mRNA表达显著上升(P<0.01),电针组无明显差异(P>0.05),电针组较模型组显著降低(P<0.01);Bcl-2/Bax及Bcl-2 mRNA/Baxm RNA的比值,模型组降低而电针组升高(P<0.01)。结论电针可以通过提升Bcl-2/Bax的比值使抗凋亡基因占据优势,从而抑制缺血再灌注区的细胞凋亡,减轻脑水肿,促进神经功能恢复。