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1.
淋巴细胞活性染色质诱导系统性红斑狼疮样小鼠模型   总被引:1,自引:0,他引:1  
目的建立活性染色质诱导系统性红斑狼疮(SLE)样小鼠模型。方法从ConA活化的BALB/c小鼠脾淋巴细胞中提取活性染色质,分别于d 0、d 14、d 21和d 28以染色质100μg在BALB/c小鼠尾根部及背部皮内注射免疫4次,诱导SLE样小鼠模型。目测半定量尿蛋白试纸法检测动物的尿蛋白变化,HE染色法检查动物的肾脏、脾脏病理改变,计算动物的胸腺和脾脏指数,MTT法检测ConA和LPS诱导的T、B淋巴细胞增殖反应,全自动生化分析仪检测血清中Crea和BUN水平,ELISA法检测小鼠血清中ANA、抗dsDNA、IgG1、IgG2a、IL-10、IFN-γ水平,流式细胞术检测脾脏T、B淋巴细胞亚群变化。结果诱导模型小鼠尿蛋白水平升高,出现肾小球肾炎、脾脏增生等病理改变;脾脏指数明显升高,LPS诱导的B淋巴细胞增殖反应增强;血清Crea、BUN、ANA、抗dsDNA、IgG1、IgG2a、IL-10和IFN-γ水平明显升高;脾脏CD19+、CD19+CD21+、CD19+CD23+、CD19+IgD+B淋巴细胞亚群百分比明显升高,CD4+CD25+T淋巴细胞百分比明显下降。结论 ConA活化淋巴细胞的染色质免疫同系BALB/c小鼠成功诱导了SLE样小鼠模型,其血清学、组织病理学及免疫学方面特征与人类SLE临床特征表现相似。  相似文献   

2.
《临床医药实践》2020,(2):131-135
目的:探讨系统性红斑狼疮(SLE)患者血清免疫功能相关指标(免疫球蛋白IgG、补体C3、补体C4)与自身抗体之间的关系。方法:选取94例SLE患者,81例其他自身免疫系统疾病患者及66例健康对照者分别作为SLE组、非SLE疾病组、健康对照组三组,检测各组血清自身抗体的表达。将SLE组按SLE疾病活动指数(SLEDAI)评分标准分成活动期组和稳定期组,按抗SmD1抗体、抗双链DNA(dsDNA)抗体阴阳性分别分成抗SmD1(+)组、抗SmD1(-)组、抗dsDNA(+)组、抗dsDNA(-)组,与健康对照组一起检测各组血清免疫球蛋白IgG、补体C3、补体C4水平。将以上各组指标进行比较分析。结果:与健康对照组相比,SLE组和非SLE疾病组自身抗体谱中各指标(除Jo-1外)阳性率差异具有统计学意义(P<0.05)。SLE组中抗SmD1抗体、抗dsDNA抗体、抗核小体抗体(AuNA)、抗组蛋白抗体(AHA)、抗核糖体P蛋白(Rib-P)抗体阳性率均比非SLE疾病组高(P<0.05)。SLE活动期组血清免疫球蛋白IgG水平高于SLE稳定期组和健康对照组,而血清补体C3,C4水平低于SLE稳定期组和健康对照组(均P<0.05),SLE稳定期组血清IgG,C3,C4水平和健康对照组相比,血清补体C3,C4水平更低,IgG水平则要高,差异有统计学意义(P<0.05)。SLE组中抗SmD1抗体、抗dsDNA抗体阳性组血清补体C3,C4水平分别低于抗SmD1抗体、抗dsDNA抗体阴性组(均P<0.05)。结论:联合检测免疫球蛋白IgG、补体C3,C4及自身抗体谱可有助于SLE的早期诊断、治疗监测及预后评估。  相似文献   

3.
白芍总苷对小鼠系统性红斑狼疮样改变的保护作用   总被引:11,自引:3,他引:8  
目的 观察白芍总苷 (TGP)对空肠弯曲菌CJ S13 1诱导的小鼠SLE样改变的保护作用。方法 采用空肠弯曲菌CJ S13 1和CFA混合免疫动物 ,诱导小鼠SLE样改变。结果 TGP 5 0 ,10 0 ,2 0 0mg·kg-1·d-1× 2 8d ,ig能部分或完全拮抗血清IgG型自身抗体水平的升高 ,抑制ConA及LPS诱导的淋巴细胞增殖反应的增强和IL 1生成的增多。结论 一定剂量的TGP对小鼠SLE样改变具有一定的保护作用  相似文献   

4.
目的优化空肠弯曲菌(CJS131)诱导系统性红斑狼疮(SEE)样小鼠模型的方法。方法小鼠随机分为5组,即正常对照组、卡介苗(BCG)对照组、结核分枝杆菌H37Ra对照组、CJS131+BCG模型组和CJS131+H37Ra模型组。在第0天免疫,在第14、21、42天加强免疫,在第36、54、61天分批称其体质量后处死。检测小鼠免疫器官指数、血清抗核抗体水平、血清总IgG水平和肾组织病理损伤程度。结果不论采用含BCG还是H37Ra的弗氏完全佐剂(FCA),给予CJS131免疫的小鼠均有SLE样综合征的表现:血清抗核抗体及总IgG水平升高,肾组织损伤。在动态检测中病变基本维持。结论CJS。在对模型组小鼠的免疫诱导中起主要作用,以BCG或H37Ra作为FCA中采用的菌株都可引起病变,通过加强免疫可维持小鼠长期病变,优化后的模型可用于SLE治疗性药物的筛选。  相似文献   

5.
雷公藤多苷对自身免疫反应鼠的疗效机制研究   总被引:3,自引:0,他引:3       下载免费PDF全文
本文观察雷公藤多苷对空肠弯曲菌CF -1 株辅以佐剂免疫小鼠产生的自身免疫综合征的作用,探讨雷公藤多苷对自身免疫病治疗的可能机制。雷公藤多苷75 及150 mg ??kg-1??d-1ig 动物在首次免疫后d 14 给药d 28采用酶联免疫吸附法测定血清抗dsDNA 抗体,[3H] -TdR 掺入法测定淋巴细胞转化,比色法测定血清溶血素,胸腺细胞增殖法测定IL -1 活性,结晶紫染色法测定TNF ???结果显示,雷公藤多苷75 及150 mg ??kg-1可降低抗dsDNA 抗体和血清溶血素水平,抑制淋巴细胞转化及腹腔巨噬细胞产生IL -1 和TNF ???小鼠后肢肿胀程度明显减轻且体重增加。提示雷公藤多苷可抑制亢进的自身免疫反应。  相似文献   

6.
陈伟华  王薇 《河北医药》2014,(8):1157-1158
目的探讨SLE患者血清免疫球蛋白及补体与自身抗体的相关性。方法采用速率散射免疫比浊法检测109例SLE患者和30例体检健康对照者的血清Ig及C3、C4水平并与自身抗体结果进行比较。结果SLE患者血清IgG、IgA水平高于对照纽(P〈0.05);血清IgM水平与对照组比较差异无统计学意义(P〉0.05);补体C3、C4水平低于对照组(P〈0.05)。ANA高滴度组与低滴度组相比较,IgG明显升高,补体C3明显降低(P〈0.01);而IgA、IgM及c4差异无统计学意义(P〉0.05)。抗Sm及抗ds-DNA抗体阳性组分别与其对应的阴性组血清Ig及c3、C4水平相比较,Ig水平差异无统计学意义(P〉0.05),而C3水平明显降低(P〈0.叭)。结论SLE患者存在血清IgG水平升高,血清补体C3、C4降低的现象;随自身抗体滴度升高差异更为显著,且IgG与疾病活动呈正相关,C3、C4与疾病活动呈负相关;C3可作为反映SLE疾病活动的参考指标。  相似文献   

7.
目的评估依鲁替尼长期给药对姥鲛烷诱导的系统性红斑狼疮(SLE)模型小鼠的治疗作用及副作用。方法 6周龄雌性BALB/c小鼠ip给予姥鲛烷0.5mL制备SLE小鼠模型。4周后,根据抗双链DNA(DS-DNA)抗体滴度和体质量均匀分为模型组、依鲁替尼30 mg·kg~(-1)治疗组和泼尼松10 mg·kg~(-1)治疗组,每天ig给药1次,连续28周。每4周测定1次体质量,ELISA法测定血清中抗DS-DNA抗体、抗单链DNA(SS-DNA)抗体和抗组蛋白抗体水平,评估关节炎红肿等症状的评分及发病率;生物化学法检测血清肌酐、尿素氮和尿液尿蛋白水平评估肾功能。给药28周后处死小鼠,ELISA检测白细胞介素6(IL-6)、干扰素γ(IFN-γ)和肿瘤坏死因子α(TNF-α)水平;采集心、肝、脾、肺和肾,称重并计算脏器系数;生物化学法检测血清谷丙转氨酶、谷草转氨酶和碱性磷酸酶活性评估肝功能;HE染色观察后足和肾组织病理变化,并用免疫组化法检测肾组织免疫复合物IgG沉积。结果与正常对照组比较,模型组抗DS-DNA,SS-DNA和组蛋白抗体及IL-6,IFN-γ和TNF-α水平均明显升高(P<0.01),血清肌酐、尿素氮和尿蛋白升高,关节炎症状严重(P<0.01),后足组织炎症细胞浸润严重(P<0.01),肾和脾显著增大(P<0.01)与模型组相比,依鲁替尼治疗28周可减缓SLE模型小鼠体质量下降,降低抗DS-DNA,SS-DNA和组蛋白抗体水平(P<0.01),减轻小鼠关节红肿等症状(P<0.01),并降低关节炎发病率,降低血清细胞因子IL-6,IFN-γ和TNF-α水平(P<0.01);后足关节病理观察显示,炎症细胞浸润、血管翳形成、软骨破坏和骨吸收减轻(P<0.01);肾组织病理观察显示,炎症细胞浸润和IgG免疫复合物沉积减少,血清肌酐、尿素氮和尿蛋白水平降低(P<0.01),血清GPT和ALP活性亦降低(P<0.01)。结论依鲁替尼给药28周对SLE模型小鼠有一定的治疗作用,未见明显副作用。  相似文献   

8.
目的 研究白芍总苷对小鼠免疫性肝损伤的保护作用。方法 在建立BCG +LPS诱导免疫性肝损伤小鼠模型的基础上 ,分光光度法检测血清中ALT、AST、NO水平和肝匀浆MDA、GSH Px、SOD含量 ;放免法检测TNF α的生物学活性 ;细胞增殖法测定脾淋巴增殖反应。结果 白芍总苷 (6 0、12 0、2 4 0mg·kg-1)ig给药可明显降低免疫性肝损伤小鼠增高的血清ALT、AST活性 ,同时能减少肝匀浆MDA含量 ,使降低的肝匀浆GSH Px、SOD活性升高 ,进一步研究发现白芍总苷可明显抑制免疫性肝损伤小鼠血清NO和TNF α的产生。白芍总苷还可抑制小鼠腹腔巨噬细胞TNF α的产生 ,对ConA诱导的脾淋巴增殖反应具有恢复作用 ,而对LPS诱导的脾淋巴增殖反应无明显影响。结论 白芍总苷对免疫性肝损伤具有保护作用。  相似文献   

9.
目的:研究槲皮素对降植烷(pristane)诱导的红斑狼疮样小鼠模型的治疗作用。方法:18只BALB/c雌性小鼠随机分为3组,即正常对照组(NC组,n=6;实验首日生理盐水0.5 mL腹腔注射);模型组(SLE组,n=6;实验首日降植烷0.5 mL腹腔注射);脂质体槲皮素治疗组(LQ组,n=6;实验首日降植烷0.5 mL腹腔注射;第二日起槲皮素按体质量降植烷50 mg·kg-1 剂量灌胃,隔日一次)。5个月治疗期结束后ELISA法测定各组小鼠血清自身抗体(ANA、抗ds-DNA和抗snRNP/Sm抗体)表达水平及采用免疫荧光法显示各组小鼠肾小球IgG蛋白沉积情况。结果:3组小鼠ANA、抗ds-DNA和抗snRNP/Sm抗体表达水平差异有统计学意义(F=51.791,39.745和30.411,P<0.01)。与SLE组相比,LQ组小鼠肾小球的IgG沉积明显减少,自身抗体表达水平显著降低,但LQ组自身抗体表达水平仍高于NC组(P<0.01)。结论:槲皮素能降低降植烷诱导的SLE样综合征小鼠模型自身抗体表达水平,并能减轻肾损伤程度,对SLE的治疗可能有一定作用。  相似文献   

10.
目的 探讨体内产生抗核抗体的抗原。方法 分别以羊红细胞、伤寒沙门菌及鸡血清免疫小鼠 ,同时设立生理盐水对照组 ,每组 10只 ,采用商品化酶联免疫法检测抗核抗体试剂盒 ,将酶结合物 (酶标抗人IgG)换成酶标金黄色葡萄球菌A蛋白 (SPA) ,建立检测抗核抗体的SPA ELISA法 ,并使其性能保持不变 ,用其检测小鼠的抗核抗体。结果 新建立的检测抗核抗体的SPA ELISA法与原法检测同一批标本 ,相关系数为 0 931,P <0 0 1。小鼠血清抗核抗体检测结果 (阳性率 ) :羊红细胞组为 90 % (9 10 ) ,伤寒沙门菌为 70 % (7 10 ) ,鸡血清组为 10 0 % (10 10 ) ,生理盐水对照组未检出抗核抗体。各实验组与对照组比较差异有显著性 (P <0 0 1)。结论 体内抗核抗体可能是一类由不同抗原刺激非特异产生的抗体。  相似文献   

11.
AIM: To establish an animal model for systemic lupus erythematosus (SLE)-like syndrome in mice. METHODS:BALB/c mice were immunized with active chromatin isolated from ConA-actived syngeneic spleno-lymphocytes.Plasma samples of mice were tested by enzyme-linked immunosorbent assays (ELISA) for the presence of IgG anti-dsDNA, -ssDNA, and anti-histone antibodies. Tumor necrosis factor-α(TNF-α) in serum was measured by ELISA. Spleno-lymphocyte proliferation assays and the levels of interferon-γ(IFN-γ) in supematants were tested respectively. Proteinuria was measured. Kidneys were examined by direct immunohistochemical method and fight microscopy. RESULTS: Anti-ds DNA, ssDNA, and histone antibodies were induced in active chromatin-immunized mice, the proliferation response of splenocytes to ConA and LPS were reduced, levels of interferon-γ in supernatants and TNF-at in serum were lowered. Lupus nephritis was assessed by the presence of Ig deposits,glomerular pathology and proteinuria. CONCLUSION: The active chromatin-induced SLE-like mouse model was similar to idiopathic SLE in human.  相似文献   

12.
The aim of this study was to determine whether tripterine, isolated from Tripterygium wilfordii Hoog f. in China, had beneficial effects on experimental systemic lupus erythematosus induced by active chromatin in BALB/c mice. BALB/c mice were immunized with active chromatin isolated from concanavalin A-activated syngenetic spleno-lymphocytes on day 0. Tripterine 6 or 12 mg kg(-1) day(-1), or prednisone 5 mg kg(-1) day(-1) was given to BALB/c mice intragastrically from day 35 to day 50. Treatment with tripterine 12 mg kg(-1) day(-1) for 15 days protected renal from glomerular injury with a concomitant reduction of serum autoantibodies and total immunoglobulin G (IgG) also with a improvement of splenocyte proliferation stimulated with concanavalin A and lipopolysaccharide. The effects were associated with reduced interleukin-10 production and serum nitric oxide (NO) level but not interferon-gamma compared with vehicle-treated control group. Tripterine 6 mg kg(-1) day(-1) had no significant protective effect against glomerular injury. It inhibited autoantibodies and interleukin-10 production but had no effect on splenocyte proliferation, serum NO level, and interferon-gamma production. These findings suggested that tripterine had a beneficial effect on systemic lupus erythematosus induced by active chromatin in BALB/c mice.  相似文献   

13.
Aim: To investigate the effects of co-delivering 11_-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pClA-P, which encodes the surface protein antigen PAc of Streptococcus mutans. Methods: Plasmid pCI-IL-6 was constructed by inserting the murine IL-6 gene into the pCI vector. Expression of IL-6 in vitro was assessed using Western blot analysis. BALB/c mice were intranasally co-immunized with pClA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA (slgA) were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-y and IL-4 production was measured using ELISA. Splenocyte proliferation was analyzed with flow cytometry. Rats were similarly immunized, and dental caries scores were determined using the Keyes method. Results: Marked expression of IL-6 was found in C0S-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and slgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-y from splenocytes in response to re-stimulation with PAc protein was significantly higher in the pCI-IL-6 co-immunized mice than that in the control mice, whereas the secretion of IL-4 had no significant difference. The proliferation of splenocytes from the pCI-IL-6 co-immunized mice was significantly higher than that from the mice immunized with pClA-P and pCl vector. In the rat caries model, the pCI-IL-6 co-immunization rats displayed lower caries scores than the control rats. Conclusion: Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.  相似文献   

14.
研制抗胃泌素释放肽(Gastrin-releasing peptide,GRP)单克隆抗体,并且初步评估其在体外对乳腺癌细胞生长的抑制作用。纯化重组的GRP融合蛋白,免疫BALB/c小鼠,取经免疫的小鼠脾细胞与小鼠骨髓细胞SP2/0进行细胞融合,对杂交瘤细胞进行筛选,阳性孔经3次有限稀释法亚克隆,获得稳定分泌GRP抗体的杂交瘤细胞株,进一步从腹水中用硫酸铵沉淀法纯化单抗,并以间接ELISA、Western免疫印迹以及亚类分型等方法进行抗体鉴定,之后在体外用MTT比色法测定该单抗抑制乳腺癌细胞EMT-6生长的作用。结果表明通过筛选获得了一株阳性杂交瘤细胞,命名为W9,这株杂交瘤细胞分泌的单抗具有效价高,特异性好的特点,其亚型为IgG1,体外的细胞实验也表明这种单克隆抗体能够显著地抑制EMT-6细胞的增殖。  相似文献   

15.
目的制备相思子毒素单克隆抗体并鉴定其特性。方法以甲醛处理的相思子毒素毒蛋白为抗原免疫BALB/c小鼠;取免疫小鼠的脾细胞与Sp2/0骨髓瘤细胞融合,经间接ELISA法筛选、融合细胞有限稀释法克隆、克隆化杂交瘤细胞株的亚类鉴定等方法筛选出单克隆抗体杂交瘤细胞株,用杂交瘤细胞株诱生小鼠腹水,应用蛋白A亲和层析法进行单抗的纯化,并对单克隆抗体的特异性进行鉴定。结果获得了4株可稳定分泌单克隆抗体的杂交瘤细胞2D3、4E6、1C8和1E5,诱生的腹水效价分别为1∶1×107、1∶1×106、1∶1×105、1∶1×106,亚类鉴定表明2D3为IgG1,其余3株均为IgG2b;特异性鉴定显示它们与多种毒素均无交叉反应,经过亲和层析,获得了纯化的单抗。结论获得了特异性的相思子毒素单克隆抗体,为建立相思子毒素的检测及纯化方法奠定了基础,其中4E6的效价最高,可作为检测相思子毒素的核心试剂。  相似文献   

16.
目的 观察弓形虫可溶性速殖子抗原(STAg)免疫小鼠后,小鼠血清IgG抗体水平的动态变化,探索STAg抗原的最佳免疫次数.方法 5~6周龄BALB/c小鼠20只,随机分成对照组和实验组,其中对照组头背部皮下注射生理盐水,实验组头背部皮下注射20 μg STAg,免疫5次,每次免疫1周后取5只小鼠,采血并分离血清,通过ELISA法测定IgG抗体;通过Western Blot方法检测血清中产生的IgG和IgM抗体条带.结果 第二次免疫后小鼠血清IgG值明显高于初次免疫小鼠的血清IgG值,差异有统计学意义;但是.Western Blot检测的IgG抗体条带颜色未明显增强.第四次免疫后小鼠血清IgG再次增高并达到峰值,同时第四次免疫后小鼠血清中的IgG抗体条带的反应颜色也明显变深.结论 利用STAg免疫小鼠,免疫2次就会产生明显的免疫效果,但是免疫4次可进一步有效提高小鼠的免疫力.  相似文献   

17.
目的:研究蟛蜞菊内酯对脂多糖(lipopo-lysaccharide,LPS)诱导RAW264.7巨噬细胞环氧化酶2(COX-2)、NO及TNF-α的作用。方法:ELISA方法检测0.2、2、20μmol/L不同浓度蟛蜞菊内酯对终浓度为10μg/mL LPS诱导RAW264.7细胞产生TNF-α、NO及前列腺素E2(PGE2)的影响,Western blot方法检测蟛蜞菊内酯对LPS诱导COX-2酶蛋白表达的影响。结果:LPS能够明显诱导小鼠RAW264.7细胞产生的COX-2酶蛋白,蟛蜞菊内酯低中高3个浓度均能抑制LPS诱导产生的COX-2酶蛋白表达。PGE2可以被LPS诱导增加,与空白组比有显著差异。蟛蜞菊内酯低中高3个浓度均能抑制LPS诱导产生的PGE2、NO和TNF-α,呈现剂量依赖性。结论:蟛蜞菊内酯抗炎的作用机制可能为抑制COX-2的蛋白表达,进而抑制PGE2的生成,也可能与抑制NO和TNF-α生成有关。  相似文献   

18.
Wang Z  Xie JY  Xu H  Cheng XQ  Yue XL  Li H  Zhang YY  Lu Y  Chen DF 《药学学报》2010,45(6):711-717
Matteuccia struthiopteris is a nature plant, which contains a lot of potential active components. In the present study, we investigated the effect of polysaccharides extracted from Matteuccia struthiopteris on lupus-like syndrome induced by Campylobacter jejuni CJ-S131 in BALB/c mice. Mice were randomly divided into normal, model control, SLE model (vehicle treated), Matteuccia struthiopteris polysaccharides treated (30 and 15 mg x kg(-1)) groups and prednisone 5 mg x kg(-1) treated groups. The effect of Matteuccia struthiopteris polysaccharides (Ms) on weight and organ index of BALB/c mice was detected. Autoantibodies and total IgG production were measured by enzyme linked immunosorbent assay. Proteinuria was measured and kidneys were examined by light microscopy. Compared with SLE model group, treatment with Matteuccia struthiopteris polysaccharides 30 and 15 mg x kg(-1) reduced weight loss and Matteuccia struthiopteris polysaccharides 15 mg x kg(-1) reduced spleen swelling (P < 0.05). The increased production of autoantibodies and total immunoglobulin G (IgG) were also significantly inhibited. Matteuccia struthiopteris polysaccharides protected kidney against glomerular injury in BALB/c mice with reduced immunoglobulin deposition and lowered proteinuria (P < 0.01). Matteuccia struthiopteris polysaccharides had a protective effect on lupus-like syndrome induced by CJ-S131 in BALB/c mice.  相似文献   

19.
姜黄素衍生物体外抗炎及防治小鼠脓毒症的研究   总被引:1,自引:0,他引:1  
目的探讨姜黄素衍生物FM0901体外抗炎作用,及其对大肠埃希菌引起的细菌脓毒症小鼠的保护作用。方法①用脂多糖(LPS)刺激RAW264.7细胞,ELISA法测定FM0901对细胞分泌TNF-α、IL-6水平的变化硝酸还原酶法测定NO的变化;②小鼠尾静脉注射大肠杆菌建立脓毒血症模型,考察FM0901对小鼠血清中TNF-α、IL-6及NO的影响,观察小鼠的活动状态及生存率。结果①FM0901显著抑制LPS诱导RAW264.7细胞的TNF-α、IL-6和NO释放,并具有明显的量效关系;②FM0901降低脓毒血症小鼠血清中TNF-α、IL-6和NO的含量,改善小鼠的状态,低剂量组提高大肠埃希菌感染小鼠的生存率。结论FM0901抑制RAW264.7细胞炎症介质释放,减轻脓毒症小鼠的炎症反应,提高其生存率。  相似文献   

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