雷公藤多甙对(口恶)唑酮诱导小鼠肠炎模型脾淋巴细胞因子的作用及其机制

目的　观察小鼠肠炎模型体外实验中 ,雷公藤多甙 (MGT)对唑酮 (oxazolone,OXZ)诱导的小鼠肠炎模型中脾脏淋巴细胞细胞因子分泌类型的影响 ,从免疫学、分子生物学角度探讨MGT治疗炎症性肠病 (IBD)的作用机制。方法　采用SJL/J小鼠 ,经直肠注入OXZ制成IBD模型 ;3d后将小鼠处死 ,即刻取出脾脏 ,收集脾细胞 ,将MGT(0 .1和 0 .0 1mg/ml两个浓度 )分别加入培养的淋巴细胞液中 ,行ELISA检测白介素 4 (IL 4 )和γ 干扰素 (IFN γ)。结果　 1.MGT对IFN γ生成的影响 :(1)正常对照组 :未加入MGT组 (空白对照 )的IFN γ为 (1.2 4± 0 .13) pg/ml;0 .0 1mgMGT组为(0 .97± 0 .2 6 )pg/ml;0 .1mgMGT组为 (0 .87± 0 .18) pg/ml;(P <0 .0 2 ) ;(2 )OXZ肠炎模型组 :与正常对照组比较 ,未加入MGT组 (空白对照 )IFN γ显著降低 [(0 .6 5± 0 .0 8) pg/ml比 (1.2 4±0 .13) pg/ml,P <0 .0 1],0 .0 1mgMGT组和 0 .1mgMGT组IFN γ均低于空白对照 [分别为 (0 .4 7± 0 .0 5 ) pg/ml;(0 .4 6± 0 .0 9) pg/ml],但差异无显著性。 2 .MGT对IL 4生成的影响 :(1)正常对照组 :未加入MGT组 (空白对照 )的IL 4为 (5 .6 5± 0 .4 8) pg/ml;MGT有显著抑制作用 [0 .0 1mg/mlMGT组为(4.97± 0 .38) pg/ml;0 .1mgMGT组为 (3.98± 0 .32     

恶唑酮结肠炎小鼠模型的建立

建立合适的动物模型有助于炎症性肠病(IBD)的研究，然而目前尚缺乏类似人类溃疡性结肠炎(UC)的动物模型。目的：建立恶唑酮诱导的小鼠结肠炎模型，并评估其在IBD研究中的价值。方法：予BALB／c小鼠皮肤涂搽0．2ml3％恶唑酮(溶解于100％乙醇中)2次致敏，5天后予0.15ml1％恶唑酮(溶解于50％乙醇中)灌肠。观察小鼠的疾病活动指数(DAI)和病变结肠的组织学改变，并测定髓过氧化物酶(MPO)活性以及肿瘤坏死因子(TNF)-α干扰素(IFN)-γ和白细胞介素(IL)-4的含量。结果：结肠炎模型小鼠的DAI、组织学损伤评分和MPO活性均较对照组有明显改变，病变结肠组织的IL-4含量显著增高，TNF-α和IFN-γ含量则基本正常；结肠炎症可持续2周左右。结论：恶唑酮诱导的结肠炎是一种IL-4介导的2型辅助性T细胞(Th2)型炎症，其组织学特征和炎症分布均类似人类UC。恶唑酮小鼠结肠炎模型可作为研究UC发病机制和评估药物疗效的有益工具。     

长期应用尼古丁对恶唑酮诱导的实验性小鼠肠炎模型的影响及其机制探讨

目的分析和探讨长期应用尼古丁对恶唑酮(OXZ)诱导的实验性小鼠肠炎的影响以及肠黏膜和脾细胞生成细胞因子的类型.方法采用BALB/c小鼠,经直肠注入OXZ制成炎症性肠病(IBD)模型;皮下注射尼古丁0.5mg@kg.@d,连续3周.将小鼠处死后,取出大肠及脾脏.病理组织学观察大肠黏膜改变.采用ELISA法测定大肠黏膜及脾细胞产生的干扰素γ(IFN-γ)和白介素-4(IL-4).采用流式细胞仪检测技术(FACScan)分析脾细胞内细胞因子IFN-γ和IL-4的表达.结果尼古丁组的病理组织学计分显著低于OXZ组(19.8比23.7,P＜0.02).与对照组比较,OXZ组大肠黏膜[(185±47)pg/g比(94±14)pg/g]和脾细胞生成的IL-4明显高于对照组[(59±12)pg/ml比(10±1)pg/ml];与OXZ组比较,尼古丁组IL-4生成量更趋降低[肠黏膜(157±38)pg/g比(185±47)pg/g,P＜0.05;脾细胞(50±13)pg/ml比(59±12)pg/ml,P＜0.05].OXZ组IFN-γ/IL-4比值明显低于对照组(黏膜1.10±0.37比3.40±0.35,P＜0.02;脾细胞275±1.90比30.70±3.90,P＜0.01),与尼古丁组差异无显著性(黏膜110±0.37比0.78±0.14;脾细胞2.75±1.90比0.78±0.40).OXZ组,表达IL-4的CD+4细胞数较表达IFN-γ的CD+4细胞数高13.6倍.尼古丁组表达IL-4及IFN-γ的CD44+细胞数仅为OXZ组的32%和39%.结论OXZ诱导的小鼠IBD肠炎模型是Th2亚型(IL-4升高)为主的免疫应答反应;长期应用尼古丁,通过抑制IL-4生成,从而减轻OXZ诱导的实验性小鼠肠炎的组织学损伤.     

硫化氢与小鼠恶唑酮结肠炎肠黏膜损伤的相关性

背景：研究发现内源性气体递质H2S可能具有抗炎活性。关于H2S在溃疡性结肠炎（UC）中的作用,目前研究结果不一。目的：探讨H2S与恶唑酮诱导的小鼠实验性结肠炎肠黏膜损伤的相关性。方法：健康雄性昆明小鼠96只随机分为正常对照组、模型组、NH2OH组、NaHS组,后三组建立恶唑酮结肠炎模型,NH2OH组和NaHS组分别腹腔注射H2S合成关键酶胱硫醚B合酶（CBS）抑制剂NH2OH和H2S供体NaHS干预3d或7d。实验期间评估疾病活动指数（DAI）。干预结束后处死小鼠,取病变结肠组织行组织学损伤评分,以ELISA法测定白细胞介素-4（IL-4）含量,以realtimeRT—PCR测定CBSmRNA表达,同时检测血浆H2S含量。结果：模型组、NH2OH组、NaHS组DAI评分、结肠黏膜组织学损伤评分和结肠组织IL-4含量、CBSmRNA表达明显高于正常对照组,NH2OH组高于模型组,NaHS组低于模型组,差异均有统计学意义（P〈0．05）。NH2OH组血浆H2S含量明显低于正常对照组和模型组,NaHS组明显高于正常对照组和模型组,差异均有统计学意义（P〈0．05）。结论：内源性H2S与小鼠恶唑酮结肠炎的肠黏膜损伤之间存在明显相关性,可能起抗炎和肠黏膜保护作用。     

长春新碱、强的松及雷公藤多甙治疗脾功能亢进的临床探讨

目的：研究药物治疗脾功能亢进（脾亢）的疗效。方法：采用长春新碱（ＶＣＲ）、强的松（Ｐｒｅｄ）及雷公藤多甙（ＴＲＩＰＴＥＲＹＧＩＵＮ ＮＵＬＴＩＧＬＹＣＯＳＩＤＯＲＵＭ）（ＶＰＴ）方案治疗脾亢患者。结果：１０例脾亢患者用药第１周均见脾脏缩小,第４周明显缩小,第６周６例脾脏肋下未触及,另４例Ｂ型超声检查脾长径平均缩小３．１ｃｍ。１０例肝门静脉（ＰＶ）直径除１例由１．６ｃｍ减到１．４ｃｍ外,余均在１．３     

雷公藤多苷对溃疡性结肠炎小鼠结肠组织IL-1、TNF-α、IL-13的影响

[目的]观察雷公藤多苷对溃疡性结肠炎模型小鼠IL-1、TNF-α、IL-13的影响.[方法]采用右旋葡聚糖硫酸钠法（DSS）复制BALB/c小鼠溃疡性结肠炎模型,雷公藤多苷灌胃给药7d后,采用ELISA酶联法检测结肠组织匀浆蛋白细胞因子（IL-1、TNF-α和IL-13）的含量.[结果]雷公藤多苷组小鼠结肠组织中IL-1、TNF-α含量明显下降（P＜0.05）,IL-13含量上升.[结论]雷公藤多苷通过影响细胞因子水平而达到抗溃疡性结肠炎的作用.     

核因子-κB和激活蛋白-1在恶唑酮诱导的小鼠实验性肠炎中的表达

目的 研究恶唑酮诱导的小鼠实验性肠炎中核因子(NF)-κB和激活蛋白(AP)-1基因表达量的变化,以探索其在发病机制中的意义.方法 7～8周龄的健康昆明小鼠24只,体重25～30 g,均分为正常组和模型组.模型组小鼠采用3%恶唑酮皮肤致敏5 d后,以0.5%恶唑酮(溶解于50%乙醇中)0.15 ml一次性灌肠造模方式建立实验性结肠炎模型.3 d后处死所有小鼠,分别提取外周血单个核细胞(PBMC)、脾脏单个核细胞(SMC)和肠黏膜固有层单个核细胞(LPMC).经细胞mRNA抽提、逆转录cDNA后进行荧光定量PCR(Taqman探针法)检测NF-κB、AP-1的表达量.并进行结肠炎的组织学评分.结果 模型组NF-κB在SMC、LPMC和PBMC中的表达量均比正常组明显增高(分别为5.62±0.78比3.16±0.59、5.46±0.38比3.18±0.58、5.65±0.56比3.36±0.59,P＜0.01),AP-1亦是如此(分别为5.61±0.54比3.22±0.50、5.50±0.69比3.19±0.40、5.67±0.44比3.27±0.41,P＜0.01).结论 NF-κB、AP-1的活化可能与结肠炎的发病机制有关.     

雷公藤多甙治疗儿童过敏性紫癜性肾炎的临床疗效及其对Th1、Th2细胞因子的影响

目的 探讨雷公藤多甙(TWP)治疗儿童过敏性紫癜性肾炎(HSPN)的临床疗效及其对Th1/Th2细胞因子的影响.方法 将107例HSPN患儿随机分为常规治疗(对照组)45例、常规治疗+TWP治疗(治疗组)62例,总疗程36周,疗程结束后观察疗效;治疗前后检测血清Th1/Th2细胞因子(IL-2、IL-4、IL-6、IL-13、TNF-α)变化.结果 治疗组疗效优于对照组(P<0.05);两组Th1/Th2细胞因子均降低(P均<0.01),但以治疗组下降明显.结论 TWP对HSPN患儿有较好疗效,其机制可能与抑制Th1/Th2细胞因子水平有关.     

康复新液对小鼠嗯唑酮结肠炎的疗效及其机制初探

我国溃疡性结肠炎的发病率不断升高,现有治疗尚存在不足,需寻找新的治疗方法和药物。目的：观察康复新液对小鼠嗯唑酮（OXZ）结肠炎的疗效,初步探讨其治疗机制。方法：将60只小鼠随机分为空白对照组、模型组、康复新液组、高浓度康复新液组、激素组,以OXZ灌肠诱导结肠炎模型。分别于灌肠后第3、8、13d处死小鼠。行疾病活动指数（DAI）、结肠大体评分和组织学评分,以ELISA法检测结肠组织髓过氧化物酶（MPO）、白细胞介素4（IL4）、干扰素．1（IFN-1）、肿瘤坏死因子．Or．（TNF—d）、NF—KB含量。结果：灌肠后第3、13d,与空白对照组相比,模型组DAI、结肠大体评分、组织学评分、MPO、IL-4、TNF-d、NF—KB含量均明显升高（P〈0．05）,IFN．1含量无明显差异。灌肠后第8、13d,给予康复新液和高浓度康复新液治疗后,上述指标均明显改善（P〈0．05）,IFN-y含量无明显差异。灌肠后第8d,激素组仅大体评分和IL-4含量明显降低（P〈0．05）。结论：康复新液灌肠可使OXZ诱导的小鼠结肠炎症明显减轻,其作用可能与抑制肠道异常炎症反应、调节细胞因子水平、抑制NF—KB转录有关。     

Insulin-modulated interleukin-2 production by murine splenocytes and a T-cell hybridoma

Murine interleukin-2 (IL-2)-producing AOFS 21 T-cell hybridoma cells and normal murine splenocytes were stimulated in serum-free media by 16 potential mitogens/growth factors. Only insulin, concanavalin A (Con A), peptidoglycan monomer and a tumour-derived insulinoid stimulated [3H]thymidine incorporation by AOFS 21 cells and splenocytes. Supernatants of these stimulated cultures were tested for IL-2 activity which generally followed the pattern of growth stimulation. Both the mitogenicity and stimulation of IL-2 secretion by insulin were second only to the effects of Con A.     

Tripterygium glycosides suppress production of interleukin‐4 by splenocytes in oxazolone‐induced murine colitis

OBJECTIVE: To investigate the in vitro effect of Tripterygium glycosides (TG) on cytokine production by splenocytes in oxazolone (OXZ)‐induced colitis in mice. METHODS: Oxazolone (6 mg in 50% ethanol) was administered to male SJL/J mice intrarectally to induce colitis and the mice were killed 3 days later. Isolated splenocytes were cultured for 24 h in the presence of phorbol myristate acetate and ionomycin. A preparation of Tripterygium glycosides at a concentration of either 0.1 mg/mL or 0.01 mg/mL was added to the culture medium of splenocytes. Production of interferon gamma (IFN‐γ) and interleukin‐4 (IL‐4) in the supernatant were measured by ELISA. RESULTS: Production of IFN‐γ in the normal control group was suppressed by TG at both concentrations (0.01 and 0.1 mg/mL; 1.24 ± 0.13 pg/mL (samples without TG) → 0.97 ± 0.26 pg/mL (0.01 mg/mL TG) → 0.87 ± 0.18 pg/mL (0.1 mg/mL TG); P < 0.02) in a dose dependent manner. In the OXZ‐induced colitis group, the basic level of IFN‐γ was significantly lower than that of the normal control group (1.24 ± 0.13 pg/mL vs 0.65 ± 0.08 pg/mL; P < 0.01); but IL‐4 production was significantly increased in the OXZ‐induced colitis without TG group (7.83 ± 0.69 pg/mL vs 5.65 ± 0.48 pg/mL, P < 0.01). In both groups, TG suppressed IL‐4 production in a dose‐dependent manner (normal control group: 5.65 ± 0.48 pg/mL (samples without TG) → 4.97 ± 0.38 pg/mL (0.01 mg/mL TG) → 3.98 ± 0.32 pg/mL (0.1 mg/mL TG), P < 0.01; OXZ group: 7.83 ± 0.69 pg/mL (samples without TG) → 7.07 ± 0.47 pg/mL (0.01 mg/mL TG) → 6.35 ± 0.48 pg/mL (0.1 mg/mL TG), P < 0.01). CONCLUSION: Oxazalone‐induced IL‐4 overproduction by splenocytes was significantly suppressed by TG in a dose dependent manner and the beneficial effects of TG on ulcerative colitis might be related to the suppression of the Th2 type (e.g. IL‐4) mediated immunological response of splenocytes.     

阿泰宁对噁唑酮诱导的大鼠结肠炎模型的治疗作用

目的:建立嗯唑酮诱导的大鼠结肠炎模型.观察阿泰宁对噁唑酮诱导的大鼠结肠炎的治疗作用及机制.方法:经直肠注入嗯唑酮建立大鼠溃疡性结肠炎模型.将大鼠随机分为:空白对照组(正常组,n=8),模型组(n=10),美沙拉秦组(正常组,n=10),阿泰宁组(n=10),治疗21d后处死动物,肉眼观察结肠病变,分别测体质量、结肠湿质量、脾脏质量和结肠组织病理学变化,ELISA法测定大鼠血清IL-1β、IL-10、TNF-α以及结肠黏液内容物sIgA含量,考马斯亮兰法测定血清总蛋白,溴甲酚绿比色法测定白蛋白以及肠道菌群培养.结果:模型组、阿泰宁组和美沙拉秦组大鼠体质量均比正常组显著降低(均P＜0.01).美沙拉秦组和阿泰宁组血清球蛋白含量、结肠湿质量指数较模型组差异显著(29.9±5.7,29.1±5.4vs23.7±9.5:6.0±0.9.6.2±0.4vs7.4±1.6,均P<0.05).模型组大鼠血清IL-1β和TNF-α含量较正常组、美沙拉秦组和阿泰宁组差异显著(44.6±17.2vs8.8±7.9,14.5±4.7,8.6±3.4,均P<0.01;33.5±7.2 vs 22.6±6.7,22.3±9.2,24.4±10.8,均P＜0.05).模型组大鼠血清IL-10含量较正常组、阿泰宁组差异显著(101.5±35.8vs280.5±36.1,271.3±33.8,P<0.01).阿泰宁组脾脏指数、sIgA含量比正常组显著升高(3.4±0.8vs2.7±0.3;46.0±20.3vs23.4±18.5,均P<0.05),而美沙拉秦组差异不显著.较模型组双歧杆菌数量,阿泰宁治疗后明显上升,梭杆菌数量明显下降(9.7±0.1vs9.3±0.2:3.7±0.3vs5.8±0.7,均P<0.01).结论:阿泰宁能够有效治疗噁唑酮诱导的大鼠结肠炎.     

Characterization of murine hematopoietic progenitor subsets involved in interleukin-3-induced interleukin-6 production

Various murine cell populations were tested for their ability to generate interleukin-6 (IL-6) in response to IL-3. Among these, bone marrow cells exhibit the most prominent IL-6 production. The responder cells in this organ have been further characterized by cell fractionation on a discontinuous Ficoll gradient, fluorescence-activated cell sorting, and in situ hybridization. These procedures have allowed us to ascribe the following features to the cells mainly responsible for IL-3-induced IL-6 production: (1) they possess a low density and a relatively high forward and perpendicular light scatter (FLS/PLS); (2) they are characterized by a high rhodamine (Rh) retention; and (3) their enrichment in various subpopulations is similar to that obtained for progenitors forming colonies in the methylcellulose assay colony-forming units (CFU-C). In contrast, IL-3 target cells in terms of IL-6 production are absent both in the mature and in the most immature bone marrow compartment. Indeed, the Rh-dull population that is enriched for cells with marrow repopulating activity does not respond to the growth factor and mature cells cannot be induced to express IL-6 as assessed by (1) FLS/PLS characteristics, (2) the monoclonal antibody ER-MP 20 recognizing monocytes and granulocytic cells, and (3) in situ hybridization. Taken together, our data support the conclusion that the bone marrow cells generating IL-6 in response to IL-3 belong to a progenitor population with enhanced mitochondrial activity, comprising probably several types of immature cells of the myeloid lineage including macrophage/granulocyte precursors.