溃结饮不同给药途径对溃疡性结肠炎大鼠前炎性细胞因子表达的影响

[目的]通过比较口服与局部灌肠给药,探讨不同给药途径治疗溃疡性结肠炎(UC)的抗炎机制.[方法]采用2,4,6-三硝基苯磺酸/乙醇法造模,将大鼠随机分为模型组、奥沙拉嗪钠对照组、溃结饮灌胃组、溃结饮灌肠组和正常组.除正常组外,在正常饲养的同时分别给予蒸馏水、奥沙拉嗪、溃结饮灌胃和溃结饮灌肠;采用免疫组化法及图像分析测定不同给药途径下,血清肿瘤坏死因子α(TNF-α)、白细胞介素8(IL-8)水平及结肠黏膜TNF-α表达的动态变化.[结果]在抑制机体整体的炎症反应方面,溃结饮灌胃或灌肠治疗均可在急性期发挥作用,在急性期的早期溃结饮灌胃的作用优于灌肠,至后期则灌肠给药下调TNF-α的作用与灌胃给药无显著差异,下调IL-8的作用优于灌胃给药;局部抗炎作用方面,急性期至慢性期溃结饮灌胃或灌肠均有作用,急性期结肠黏膜TNF-α表达灌肠组显著低于灌胃组(P＜0.05).[结论]溃结饮灌肠给药整体抗炎作用的发挥高峰在时序上迟于灌胃给药,在抑制愈后复发方面远期效果优于灌胃给药.局部抗炎作用方面,急性期灌肠的作用优于灌胃给药,至慢性期两者无显著差异.     

活血健脾补肾法对结肠炎小鼠结肠组织TNF-α及其mRNA表达的影响

目的:观察TNF-α及其mRNA在结肠炎小鼠结肠组织的表达,探讨活血健脾补肾法方药对其影响．方法:用右旋葡聚糖硫酸钠(DSS)诱导小鼠产生溃疡性结肠炎模型．将小鼠随机分为空白对照、模型、柳氮磺胺吡啶(SASP)、活血法、健脾法及补肾法组;免疫组化、原位杂交法分别观察治疗后小鼠结肠组织TNF-α及其mRNA的表达．结果:模型组小鼠结肠组织学损伤积分(炎症、病度深度、隐窝破坏)分别为4．85±2．1,5．77±2．2,7．76±2．4;活血法组 (2．24±2．4,2．53±2．5,3．49±2．3)、健脾法组(2．76 2．2,2．89±2．4, 3．87±2．3)、补肾法组(2．12±2．3,2．33±2．2,3．44±2．4)与模型组比较有显著意义(4．85±2．1,5．77±2．2,7．76±2．4)(均 P<0．05)．模型组小鼠结肠组织TNF-α及其mRNA的表达(染色积分、平均光密度、平均灰度)分别是6．8±1．4,0．35±0．03． 78．6±4．4;活血法组(3．7±1．1,0．18±0．05,137．9±6．7)、健脾法组(3．4±1．3,0．16±0．03,155．1±8．8)、补肾法组(3．1± 1．5,0．17±0．04,145．6．2±7．6)与模型组比较有显著意义(6．8± 1．4,0．35±0．03,78．6±4．4)(均P<0．05)．结论:TNF-α参与DSS诱导小鼠溃疡性结肠炎的形成,活血、健脾及补肾法方药通过下调TNF-α的表达发挥防治作用．     

姜黄素对DSS诱导的小鼠溃疡性结肠炎的疗效

目的:观察姜黄素对葡聚糖硫酸钠(DSS)诱导的小鼠溃疡性结肠炎的疗效,并探讨其作用机制.方法:选♀BALB/c小鼠60只,随机分为4组,每组15只.A组:正常对照组:B组:DSS结肠炎组,小鼠每日自由摄取5%DSS溶液:C组:姜黄素治疗组,小鼠自由摄取5%DSS溶液,每日腹腔注射姜黄素悬液30 mg/kg;D组:地塞米...     

肿瘤坏死因子-α、白细胞介素-6与溃疡性结肠炎的关系

部分研究显示细胞因子改变在溃疡性结肠炎(UC)的发生机制中可能起重要作用.不同的细胞因子产物与基因启动子、信号序列、基因内含子中的单核苷酸多态现象有关.目的:测定UC患者肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的基因多态性及其血清浓度,从而探讨TNF-α、IL-6与UC的关系.方法:以聚合酶链反应-序列特异性引物(PCR-SSP)法检测60例UC患者和60名正常对照者的TNF-α、IL-6基因型,以酶联免疫吸附测定(ELISA)检测血清TNF-α、IL-6浓度.结果:UC患者TNF-α-308、IL-6-174位点基因型频率和等位基因频率与正常对照组相比均无明显差异;UC病变位于全结肠者的TNF-α-308GG基因型频率显著高于左半结肠组和直肠组(P<0.05).UC患者血清TNF-α、IL-6浓度显著高于正常对照组(P<0.05);UC患者不同基因型间血清TNF-α和IL-6浓度相比无明显差异,而正常对照组之间差异有统计学意义(P<0.05).结论:TNF-α-308、IL-6-174位点基因多态性与UC发病的易感性无关;TNF-α-308GG基因型可能与UC病变范围有关;健康人中TNF-α、IL-6基因型是其血清水平的决定性因素,而UC患者血清TNF-α、IL-6含量可能还受其他因素的影响.     

溃疡性结肠炎患者血清TNF-α、IL-8的检测及意义

目的探讨TNF-α、IL-8在溃疡性结肠炎（UC）发病中的作用及临床意义。方法采用ELISA法测定38例活动期（病情为轻度12例、中度15例、重度11例）、22例缓解期UC患者（观察组）及20例查体健康者（对照组）血清TNF-α及IL-8水平。结果观察组活动期患者TNF-α和IL-8水平均显著高于缓解期和对照组（P均〈0．05）,缓解期患者显著高于对照组（P〈0．05）。轻、中、重度患者血清TNF-α和IL-8水平逐渐升高（P〈0．05）。结论TNF-α和IL-8参与了UC的炎症过程;血浆TNF-α和IL-8水平可作为判断UC患者病变严重程度和复发的重要指标。     

兰术四草化浊解毒方对溃疡性结肠炎大鼠细胞因子水平的影响

目的探讨兰术四草化浊解毒方对溃疡性结肠炎大鼠细胞因子水平的影响。方法将60只Wistar大鼠随机分为空白组、模型组、阳性药组、兰术四草化浊解毒方大、中、低剂量组,每组10只。除空白组外,其余5组制备溃疡性结肠炎模型。造模后空白组、模型组予蒸馏水灌胃,其余各治疗组给予相应药物灌胃,连续治疗2 w。观察各组大鼠疾病活动指数、结肠黏膜组织病理,血清肿瘤坏死因子(TNF)-α,白细胞介素(IL)-8,IL-10的变化。结果与模型组比较,兰术四草化浊解毒方能够改善结肠黏膜病理变化,减低疾病活动指数评分,降低血清TNF-α及IL-8的含量,升高血清IL-10的含量(P0.05)。与阳性药组比较,兰术四草化浊解毒方大剂量组能够降低血清TNF-α及IL-8的含量,升高血清IL-10的含量(P0.05);兰术四草化浊解毒方中低剂量组能够降低血清TNF-α及IL-8的含量(P0.05),但血清IL-10含量无显著差异(P0.05)。结论兰术四草化浊解毒方治疗溃疡性结肠炎的作用机制可能与降低血清TNF-α及IL-8,升高IL-10的含量有关。     

益生菌对溃疡性结肠炎模型大鼠肠黏膜Toll样受体表达的影响及其意义

目的探讨益生菌对葡聚糖硫酸钠诱导的溃疡性结肠炎模型大鼠肠黏膜Toll样受体(TLR)的影响及其临床意义。方法 40只SD大鼠随机分为正常对照组、模型组、益生菌组和美沙拉嗪组。对照组正常饲养,不予任何处理;模型组、益生菌组和美沙拉嗪组给予含5%葡聚糖硫酸钠饮用水10 d建立溃疡性结肠炎模型,益生菌组和美沙拉嗪组于第11天分别给予酪酸梭菌活菌和美沙拉嗪灌胃治疗14 d。14 d后处死所有大鼠,观察疾病活动指数,进行组织学评分,RT-PCR法检测TLR2和TLR4的表达,免疫组化方法检测转转录因子(NF)-κB的活性。结果模型组的疾病活动指数和组织学评分显著高于对照组,益生菌组和美沙拉嗪组低于模型组(P<0.05)。模型组TLR2和TLR4的表达显著高于对照组,益生菌组和美沙拉嗪组高于对照组且低于模型组(P<0.05),而益生菌组和美沙拉嗪组间差异不显著(P>0.05)。模型组NF-κB的活性显著高于对照组,益生菌组和美沙拉嗪组高于对照组且低于模型组(P<0.05),而益生菌组和美沙拉嗪组间差异不显著(P>0.05)。结论溃疡性结肠炎模型大鼠中TLR表达异常,NF-κB的活性增加,益生菌及美沙拉嗪制剂可降低溃疡性结肠炎模型大鼠TLR表达和NF-κB的活性,是治疗溃疡性结肠炎的有效方法。     

行气活血化瘀法对溃疡性结肠炎患者血清TNF-α和IL-6的影响

目的探讨行气活血化瘀法对溃疡性结肠炎(UC)患者细胞因子的影响及疗效。方法将42例UC患者随机分成对照组20例、治疗组22例。对照组给予美沙拉嗪肠溶片口服;治疗组在对照组的基础上同时给予化瘀宁肠愈疡方灌肠,治疗4 w后,观察临床疗效,并检测治疗前后患者血清中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6的含量。结果治疗组疗效明显优于对照组(P<0.05),治疗组更能显著降低UC患者血清TNF-α、IL-6的水平。结论行气活血化瘀法治疗UC疗效显著,其调节UC患者细胞因子的作用,可能是该病的治疗机制之一。     

溃疡性结肠炎患者血清IL-6、IL-8、TNF-α水平变化及意义

目的观察溃疡性结肠炎（UC）患者血清IL-6、IL-8、TNF-α水平变化,并探讨其意义。方法 UC患者60例（UC组）,对照组30例。采用ELISA法检测两组血清IL-6、IL-8、TNF-α。结果 UC组血清IL-6、IL-8、TNF-α水平明显高于对照组（P均〈0.05）;重度UC患者血清IL-6、IL-8、TNF-α水平高于轻中度、中度高于轻度（P均〈0.05）。结论 UC患者血清IL-6、IL-8、TNF-α水平升高,检测血清IL-6、IL-8、TNF-α有助于UC病情的判断。     

四神丸对溃疡性结肠炎模型大鼠血清白细胞介素-1β、-4、肿瘤坏死因子-α的影响

目的观察四神丸对溃疡性结肠炎(UC)大鼠白细胞介素(IL)-1β、-4肿瘤坏死因子(TNF)-α的影响。方法 Wistar大鼠40只分为四组,采用2、4、6-三硝基苯磺酸(TNBS)乙醇灌肠法造模,光镜观察结肠组织病理形态并评分,采用酶联免疫法(ELISA)检测大鼠血清IL-1β、TNF-α、IL-4的水平。结果模型组大鼠结肠组织黏膜层可见溃疡形成,且有明显的炎症、充血存在,证实造模成功。模型组大鼠血清IL-1β、TNF-α含量均高于空白组(P0.05),而IL-4含量低于空白组(P0.01);治疗后,四神丸组血清IL-1β、TNF-α水平均较模型组降低(P0.05、P0.01),而IL-4含量却显著升高(P0.01)。结论四神丸能够通过下调IL-1β、TNF-α的表达,升高IL-4的表达,调节肠道异常免疫反应、抑制炎症反应,修复溃疡,进而治疗UC。     

Effect of enteral nutrition on dextran sulfate sodium-induced colitis in rats

OBJECTIVE: To study the effect of enteral nutrition (EN) on dextran sulfate sodium (DSS)‐induced colitis in rats. METHODS: Eighty‐four Sprague–Dawley rats were divided into 7 groups (12 rats in each group). The blank control group was given ordinary laboratory feed and drinking water. The experimental groups received 5% DSS as drinking water for 7 days. Of the experimental groups, the model control group received ordinary laboratory feed, protein based enteral nutrition (PEN) was fed in the PEN group, while other groups received ordinary laboratory feed plus 5‐aminosalicylic acid (5‐ASA), methyl‐prednisolone, Lactobacillus or glutamine, respectively. On the 8th day, all the rats were sacrificed. Inflammatory scores were assessed from colonic mucosa. Blood culture from inferior vena cava, fecal culture and secretary immunoglobulin‐A (S‐IgA) levels from colonic contents were determined. RESULTS: Colon inflammatory scores of Lactobacillus, PEN, glutamine and drug‐treated groups were lower than that of the model control group (P < 0.01). The ratios of bacteria translocation in the EN (PEN, Lactobacillus and glutamine) groups were lower than that in the model control group (P < 0.0083). Fecal Lactobacilli in the Lactobacillus and glutamine groups were higher than that in the model control group (P < 0.05). S‐IgA levels in colonic contents of the PEN and 5‐ASA group were lower than that in the model control group (P < 0.05). CONCLUSIONS: EN is an effective therapy for treating DDS‐induced colitis. EN could alleviate damage, promote the repair of colonic epithelial cells and inhibit bacterial translocation. Lactobacillus and glutamine could also increase the Lactobacilli in colon.     

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.     

Hemodynamics in the colonic mucosa of rats with dextran sulfate-induced colitis in the early phase

We investigated hemodynamics in the colonic mucosa of rats with experimental colitis induced by the administration of dextran sulfate sodium (DSS). As parameters of hemodynamics, we determined the indices of mucosal hemoglobin concentration (IHb) and mucosal oxygen saturation (ISO₂), measured by reflectance spectrophotometry, and an index of colonic mucosal blood flow (Flow), measured by laser-Doppler flowmetry. In the ascending colon, each parameter was measured by a combination of these methods after 1, 3, 5, 7, and 10 days of DSS administration. Histopathological examination was also performed. IHb in the DSS group increased with time; on the 7th day, the value was 126.9±8.32, while that in the control group was 85.0±4.14, IHb in the DSS group being significantly increased (P<0.02). ISO₂ in the DSS group was lower than that in the control group, and on the 7th day, was significantly lower in the DSS group (25.7±1.34) than in the control group (33.4±1.77) (P<0.01). No changes in Flow were observed in either the DSS or the control group during the administration period, and no significant difference in Flow was found between the two groups. On histopathological examination, we observed a time-dependent increase in the infiltration of inflammatory cells in the ascending colon of rats treated with DSS, but changes such as erosion and ulceration were not found in the superficial layer of the mucosa. No histopathological changes were found in the control animals. In the early phase of the experimental colitis, hemodynamic alterations in the colonic mucosa were already present at the time the slight histopathological changes developed. These observations seemed to indicate the involvement of hemodynamic alterations in the subsequent tissue injury.     

Therapeutic effect of nimesulide on colorectal carcinogenesis in experimental murine ulcerative colitis

BACKGROUND: Patients with ulcerative colitis (UC) exhibit an increased risk for the development of cancer of the colon and rectum. Cyclooxygenase (COX)-2 inhibitors are known to suppress sporadic colorectal cancer, but it is unknown whether selective COX-2 inhibitors exhibit a preventive effect in UC-associated neoplasia. This study investigated the preventive effect of nimesulide, a selective COX-2 inhibitor, on colorectal carcinogenesis in an experimental model of murine UC. METHODS: Chronic colitis was induced in mice by administration of four cycles of dextran sulfate sodium (DSS) (each cycle: 5% DSS for 7 days and then distilled water for 14 days). The mice were killed 120 days after the completion of the fourth cycle. The mice were divided into the following five groups: group A served as a disease control; group B received a diet mixed with 400 p.p.m. of nimesulide during the whole period; group C received nimesulide during the four cycles of DSS administration (active phase); group D received nimesulide for 120 days from the end of the fourth cycle (remission phase); group E received no agents including DSS and served as a normal control. RESULTS: The incidence of dysplasia and/or cancer was 28%, 15%, 11.8%, 6.7% and 0% in groups A-E, respectively. In group D, nimesulide significantly suppressed the occurrence of dysplasia and/or cancer (P < 0.05). Strong COX-2 expression was detected by immunohistochemistry in cancer and dysplastic lesions while diffusely weak COX-2 expression was also found in the residual colon (i.e. lesion-free colon). The mucosal concentration of prostaglandin E(2) was significantly lower in groups B and D than in group A. CONCLUSIONS: The administration of the selective COX-2 inhibitor nimesulide (especially during the remission phase) exerts a suppressive effect on the development of dysplasia and/or cancer in a murine model of DSS-induced colitis. These findings may have relevance to long-standing UC in humans.     

Effects of germinated barley foodstuff on dextran sulfate sodium-induced colitis in rats

Germinated barley foodstuff (GBF), derived from the aleurone and scutellum fractions of germinated barley, is rich in glutamine and low-lignified hemicellulose, and increases mucosal protein, RNA, and DNA content in the intestine when fed to normal rats. The aim of this study was to evaluate the effects of feeding GBF or germinated gramineous seeds on experimental ulcerative colitis. Sprague-Dawley rats that received 3% dextran sulfate sodium in their diets were used as an experimental colitis model. The effects of sulfasalazine, a drug used to treat inflammatory bowel disease, were compared with those of GBF. After rats had consumed diets containing GBF or various aleurone and scutellum fractions, mucosal damage; the content of mucosal protein, RNA, and DNA in the colo-rectum; and serum interleukin-8 and α₁-acid glycoprotein levels were assessed. GBF and germinated seeds more effectively prevented bloody diarrhea and mucosal damage in colitis compared with controls and rats receiving sulfasalazine, but non-germinated samples did not have a protective effect. GBF increased mucosal protein and RNA content in the colitis model. The consumption of GBF appears to prevent inflammation in a colitis model, and its effect seems to be related to the germination process. GBF and germinated seeds have the potential to serve as nutritional therapy for ulcerative colitis. (Received Mar. 17, 1997; accepted July 25, 1997)     

Dysplasia and carcinoma development in a repeated dextran sulfate sodium-induced colitis model

BACKGROUND: As an important mechanism underlying the increased risk of colorectal carcinoma development in patients with long-standing ulcerative colitis, promotion as a result of the regenerative process has been proposed. In the present study, a dysplasia-carcinoma sequence in a novel repeated colitis model in mice is documented. METHODS: Repeated colitis was induced by nine administration cycles of 3% dextran sulfate sodium (DSS; molecular weight, 54 000): each administration cycle comprised 3% DSS for 7 days followed by distilled water for the subsequent 14 days, to give conditions similar to the clinically observed active and remission phases in humans. RESULTS: Multiple colorectal tumors (nine low- and four high-grade dysplasias and two carcinomas) developed in 25 mice. These neoplastic lesions consisted of tubular structures, presenting as various types of elevated, flat and depressed tumor, similar to those in ulcerative colitis patients. A time-course study with assessment of the severity of colitis and in vivo bromodeoxyuridine uptake during a single 3% DSS administration cycle revealed a high level of regenerative activity in the colitis-affected mucosal epithelia. CONCLUSION: Thus, with the present repeated colitis model, regeneration and neoplastic lesions were apparent, the biological features of which provide evidence of a colorectal dysplasia-invasive carcinoma sequence in ulcerative colitis.     

Changes of colonic vasoactive intestinal peptide and cholinergic activity in rats with chemical colitis

The vasoactive intestinal peptide concentration was examined in the colonic wall and portal venous plasma of rats with chemical colitis by radioimmunoassay, and the colonic localization was determined with immunocytochemistry. Colonic acetylcholine esterase activity was also measured, and the response of vasoactive intestinal peptide to acetylcholine administration was determined. Colitis was induced by administration of dextran sulfate for three months. The chemical colitis was histologically similar to active human ulcerative colitis. We observed a significant increase of immunostained neurons and nerve fibers and a significant rise in the colonic wall vasoactive intestinal peptide content in chemical colitis rats, while plasma concentrations of the peptide did not change significantly. Colonic acetylcholine esterase activity was significantly elevated in colitis rats compared with control rats. Systemic administration of acetylcholine significantly increased the colonic and plasma vasoactive intestinal peptide concentrations in colitis rats. These findings demonstrated a positive association between colitis activity and an increase of vasoactive intestinal peptide and suggested that increased vagal tone promoted the peptide's release.     

Compensatory response of colon tissue to dextran sulfate sodium-induced colitis

Depletion of goblet cells (the main mucin-producing cells in the colon) is one of the most reliable histological characteristics of ulcerative colitis, whereas a major symptom of this disease is bloody diarrhea containing a large amount of mucus. The discrepancy between these phenomena was investigated in a time-course study in rats with experimental colitis induced by treatment with oral dextran sulfate sodium (DSS) for 1, 3, or 5 days. Biochemical analysis showed a reduction in mucin content in the distal side of the colon that was proportional to the duration of DSS administration. In the proximal side of the colon, however, there was a significant increase in mucin content already on the first day of treatment with DSS. This increase in colonic mucin content continued for the 5 days of treatment. In the distal side, both sulfomucin and sialomucin decreased proportionally to the duration of DSS administration. In the proximal side, there was an increase in high iron diamine-Alcian blue-positive mucins, and confirming the proliferation of goblet cells. The proliferated glands were predominantly sialylated. Goblet cell depletion and an increase in mucin production occurred in different parts of the colon. This phenomenon may be a type of compensatory function of colon tissue in response to the localized decrease of mucin production in certain portions of the colon. (Received Sept. 7, 1998; accepted Nov. 27, 1998)     

Histochemical study of colonic cancer in experimental colitis of rats

A reliable test for premalignant lesions in the development of colonic cancer in chronic ulcerative colitis has been needed. Thus, we designed this cytochemical study, using a model of experimental colitis and colonic tumors induced in Wistar male rats by the feeding of dextran sulfate sodium. The colitis had histologic similarities to ulcerative colitis in man. The percent frequency of polypoid lesions (dysplasia or dysplasia with carcinomain situ) in the cecum and ascending colon was about 25% at three months and 90% at six months of dextran sulfate feeding. The cytochemical findings by high-iron diamine-Alcian blue staining andUlex europeus agglutinin binding were chronologically paralledled by histological changes in the colonic mucosa, and the binding pattern of peanut agglutinin was not different between normal and dextran-treated animals. Moreover, some cytochemical changes that occurred during the inflammatory responses were not present in dysplastic or malignant lesions. Thus, the histochemical tests were not useful for detecting of premalignant lesions earlier than by conventional histology. Nevertheless, the dextran sulfate model of colitis in the rat appears suitable for study of cancer development in ulcerative colitis.