白藜芦醇调节STAT3与miR-21表达抗肝癌作用的初步研究

目的初步探讨白藜芦醇(Resveratrol,Res)的抗肝癌作用及其分子机制。方法建立移植性肝癌小鼠模型,口服给予Res进行干预,分析肿瘤生长状况。然后采用Real-time PCR、RT-PCR等方法检测转录因子Foxp3及微小核糖核酸-21(miR-21)的基因表达,Western blot分析信号转导子与转录激活子3(STAT3)的活性表达,流式细胞术观察CD4+CD25+T细胞数量的变化。结果与荷瘤对照组相比,Res给药组CD4+CD25+T细胞数量和Foxp3表达均明显降低,同时miR-21表达减少,该效应或许与其下调STAT3磷酸化活性水平密切相关。结论 Res可能通过调控STAT3信号分子及miR-21表达,抑制调节性T细胞产生,改变肿瘤微环境,从而发挥抗肝癌作用。     

刺松藻多糖对肝癌Hca-F荷瘤小鼠的抑瘤作用

目的研究刺松藻多糖(CFPS)对小鼠Hca-F肝癌的生长抑制作用及机制。方法建立Hca-F实体型荷瘤小鼠模型,腹腔注射不同浓度的CFPS连续12d。称重法检测CFPS对肿瘤生长、脾脏和胸腺指数的影响;采用MTT法、ELISA法分析CFPS对荷瘤小鼠脾巨噬细胞吞噬活性、细胞因子分泌水平等免疫指标的影响。结果刺松藻多糖对小鼠实体瘤生长具有抑制作用,CFPS 200 mg·kg~(-1)·d~(-1)组与对照组相比有显著性差异(P<0.05);试验组小鼠脾指数和胸腺指数高于对照组。CFPS可增强脾巨噬细胞吞噬活性,100 mg·kg~(-1)·d~(-1)以上时可促进IL-2的产生。结论 CFPS对Hca-F肝癌小鼠肿瘤生长有抑制作用,可能与提高荷瘤小鼠细胞和分子免疫活性有关。     

外源性LTB4对CIA小鼠Treg/Th17脾细胞分化的作用

目的探讨外源性白三烯B4(LTB4)对胶原诱导型关节炎(collagen-induced arthritis,CIA)小鼠脾细胞调节性T细胞(Treg)和Th17细胞分化的调节,进一步阐明LTB4在类风湿关节炎(RA)发病中的作用机制。方法建立CIA小鼠模型,取造模d28的脾细胞,体外实验分析外源性LTB4对Treg和Th17细胞分化的影响。应用流式细胞术检测CD4+CD25+Foxp3+细胞的数量,荧光定量PCR技术检测调控Treg和Th17细胞分化的特异性转录因子Foxp3和RORγt的mRNA的表达,酶联免疫吸附(ELISA)方法检测培养细胞上清IL-17的含量。结果成功建立CIA小鼠模型;造模d28分离小鼠脾细胞,体外培养加入鸡Ⅱ型胶原(CⅡ)共同孵育,随着LTB4浓度增加(0.01、0.1、1μmol·L-1),CD4+CD25+Foxp3+细胞数量相应减少,Foxp3 mRNA的表达相应降低;相反,IL-17的含量相应增加,RORγt mRNA的表达相应升高。结论LTB4抑制CIA模型脾细胞Treg细胞的分化,促进Th17细胞的分化,提示LTB4在CIA发病过程中具有一定的促炎活性。     

美洲大蠊多肽PAP-2对H22荷瘤小鼠的抑瘤作用研究

目的:探讨美洲大蠊多肽PAP-2对H22荷瘤小鼠的抑瘤作用。方法:通过小鼠腋窝皮下注射H22肝癌小鼠腹水构建荷瘤小鼠模型。将70只小鼠随机分为模型组(生理盐水)、5-氟尿嘧啶组(阳性药对照,20 mg/kg)以及美洲大蠊提取物的脱脂膏组(200 mg/kg,以提取物量计)、CⅡ-3组(从浸膏中提取的以多肽为主要活性成分的组分,200 mg/kg,以提取物量计)和多肽PAP-2高、中、低剂量组(从CⅡ-3中分离得到,200、100、50 mg/kg,以单体量计),每组10只。5-氟尿嘧啶组小鼠隔天腹腔注射给药1次,其余各组小鼠均每天灌胃给药1次,给药周期均为10 d。给药结束后,观察小鼠瘤块变化并测定其脏器(脾、胸腺、肝)指数,苏木精-伊红染色后观察其瘤组织病理变化,并采用酶联免疫吸附试验法测定其血清中血管内皮生长因子(VEGF)、白细胞介素1β(IL-1β)和IL-4的含量。结果:脱脂膏、CⅡ-3和不同剂量的PAP-2均能不同程度地抑制荷瘤小鼠瘤块的生长并升高其脏器指数,且PAP-2的作用具有一定的量效关系。其中,PAP-2高剂量组的抑瘤率(38.95%)显著高于脱脂膏组和CⅡ-3组(P<0.05),并与5-氟尿嘧啶组(40.87%)接近(P>0.05);PAP-2高剂量组小鼠脾指数、胸腺指数、肝指数较模型组均显著升高(P<0.05),并显著高于CⅡ-3组(P<0.05),且其肝指数显著高于脱脂膏组(P<0.05)。此外,PAP-2还可降低荷瘤小鼠血清中VEGF和IL-4含量、升高其血清中IL-1β含量,其中PAP-2高剂量组与模型组比较差异均有统计学意义(P<0.05);并且PAP-2高剂量组小鼠血清中IL-1β含量显著高于脱脂膏组和CⅡ-3组(P<0.05),IL-4含量显著低于脱脂膏组和CⅡ-3组(P<0.05),但是其血清中VEGF含量显著低于脱脂膏组和CⅡ-3组(P<0.05)。结论:美洲大蠊多肽PAP-2对H22荷瘤小鼠具有一定的抑瘤作用,可升高其脏器指数,降低其血清中VEGF、IL-4含量并升高其血清中IL-1β含量。     

枸杞子多糖对小鼠白细胞介素-2活性的增强作用(英文)

本文研究了中药枸杞子的提取物枸杞子多糖(LBP)对成年(2月龄)和老年(16月龄)小鼠脾白细胞介素-2(IL-2)活性的影响。在诱导脾细胞IL-2的培养液(脾细胞5×10~6/ml和ConA 4μg/ml)中加入LBP,所制备的含IL-2上清液使成年小鼠胸腺细胞在体外增殖活性([~3H]TdR参入法)升高。老年小鼠IL-2促进淋巴细胞增殖水平显著低于成年。LBP可使老年小鼠IL-2的这种作用显著提高,达成年水平。这些结果表明,LBP对IL-2的活性有增强作用并使老年小鼠IL-2活性得到恢复。     

辛伐他汀对变态反应性脑脊髓炎小鼠INF-γ和IL-10表达的影响

目的研究辛伐他汀(Simvastatin)对实验性变态反应性脑脊髓炎(experiment allergic en-cephalomyelitis,EAE)小鼠肿瘤坏死因子-γ(INF-γ)和白细胞介素-10(IL-10)表达的影响,探讨该药物对EAE保护作用的免疫调节机制。方法通过皮下注射MOG33-55肽段诱导小鼠急性EAE模型,实验分为EAE给药组、EAE对照组和正常对照组。观察各组小鼠体重等变化,测定EAE发病高峰期血清、培养的单核细胞及脾细胞培养上清液的INF-γ和IL-10水平。结果辛伐他汀能降低EAE小鼠的发病率,减轻了症状的严重程度,并能显著降低EAE小鼠血清、单个核细胞及脾细胞培养上清中INF-γ水平,升高IL-10水平。结论辛伐他汀可能通过其抑制INF-γ和上调IL-10的表达,调节免疫状态,从而对小鼠EAE起到保护作用。     

参麦注射液对荷 H22小鼠免疫功能的影响

目的研究参麦注射液对荷瘤小鼠免疫功能的影响。方法建立H22细胞小鼠移植瘤模型,将荷瘤小鼠随机分为模型组（荷瘤空白组）,参麦注射液三个剂量组,另设空白参考组,每日灌胃给药,通过测定对白细胞总数及分类,sIL-2R、IL-2、IL-4含量,脾淋巴细胞增殖及迟发超敏反应能力的影响,探讨参麦注射液的免疫调节作用。结果参麦注射液可提高荷瘤小鼠白细胞总数;降低荷瘤小鼠血清sIL-2R的含量,提高IL-2,IL-4水平,改善小鼠脾淋巴细胞增殖和迟发型超敏反应能力。结论参麦注射液具有增强机体免疫功能的作用。     

莪术醇磷酸酯单钠抗肿瘤实验研究

本文研究了莪术醇磷酸酯单钠(CP-Na)的抗肿瘤作用。CP-Na 10~(-4)-5×10~(-4)g/ml 具有杀死小鼠肝癌细胞作用。CP-Na5×10~(-4)g/ml 浓度预先在体外与艾氏腹水癌细胞作用后,对癌细胞在体内增殖有抑制作用。CP-Na 腹腔注射127mg/kg×7,对艾氏腹水癌和肝癌细胞在体内增殖均有抑制作用,CP-Na 皮下注射127mg/kg×14,对肉瘤细胞在体内增埴有抑制作用.CP-Na 腹腔注射在小鼠 LD_(50)为635.1mg/kg。     

独角莲醇提液对H22 肝癌荷瘤小鼠血清IL-2和TNF-α水平的影响

目的探讨独角莲醇提液对H22肝癌荷瘤小鼠血清IL-2和TNF-α水平的影响。方法建立体内H22肝癌荷瘤小鼠的模型，采用灌胃法给予其100mg·kg^-1·d^-1独角莲醇提液，以生理盐水灌胃和5-FU腹腔注射作为对照，连续10d，停药后第2天摘取小鼠眼球取血后断髓处死，取出胸腺、脾脏称重，计算小鼠的胸腺重量指数、脾脏重量指数，采用ELISA法检测小鼠血清中细胞因子白介素-2（IL-2）、肿瘤坏死因子-α（TNF-α）的水平。结果与生理盐水灌胃的H22肝癌荷瘤小鼠比较，5-FU腹腔注射和独角莲醇提液灌胃处理的H22荷瘤鼠血清中细胞因子IL-2、TNF-α水平明显升高；但胸腺重量指数、脾脏重量指数显著下降，差异均具有统计学意义（P〈0．05）。结论独角莲醇提液可提高H22荷瘤鼠的血清IL-2和TNF-α水平。     

胀果甘草多糖佐助的树突状细胞疫苗对H22肝癌荷瘤小鼠的免疫治疗作用

目的：制备胀果甘草多糖佐助的树突状细胞（dendritic cell,DC）疫苗,研究其对H22肝癌荷瘤小鼠的免疫治疗作用。方法：从小鼠骨髓中分离培养得到骨髓源树突状细胞（bone marrow-derived dendritic cells,BMDC）,用H22肝癌抗原致敏DC,再加入一定浓度的胀果甘草多糖佐剂制备DC疫苗,用倒置显微镜观察DC形态变化、流式细胞仪检测DC疫苗表型。建立H22肝癌荷瘤小鼠模型,按数字表法随机分为模型组、阳性组、DC组、TNF-α组、GiP组和GiP-B1组,用胀果甘草多糖佐助的DC疫苗免疫治疗荷瘤小鼠2次。末次给药后第5天处死小鼠,检测脏器指数及抑瘤率;酶联免疫法检测小鼠血清中IL-12(P70)、IFN-γ、IL-4和IL-10细胞因子含量;HE染色观察小鼠肝脏和肿瘤组织病理形态变化;免疫组化检测肿瘤组织Bax和Bcl-2蛋白的表达水平。结果：胀果甘草多糖作为DC疫苗佐剂,能够上调DC表面标志分子的表达水平（P<0.05）;胀果甘草多糖佐助的DC疫苗能够减缓肝癌小鼠肿瘤生长速度,升高肝癌小鼠脾脏指数和胸腺指数（P<0.05）,增加肝癌小鼠...     

Role of H1 receptors in histamine-mediated up-regulation of STAT4 phosphorylation

Histamine shifts TH1/TH2 cytokine balance from TH1 to TH2 cytokines and regulates the function of lymphocytes after binding to histamine receptors. The phosphorylation of STAT factors and the translocation to the nucleus are important steps in the regulation of TH1/TH2 cytokine balance. This study was designed to investigate the effects of histamine on the phosphorylation of STAT4. C57BL/6 splenocytes were isolated and treated with histamine (10(-4) to 10(-9) M) after activation with either PMA (phorbol 12 myristate 13-acetate) plus ionomycin or IL-12. The phosphorylated STAT4 levels were analyzed by Western Blot Analysis. Unstimulated splenocytes expressed both STAT4 and phosphorylated STAT4. However, phosphorylated STAT4 gradually declined within 24 h. Histamine increased the phosphorylation of STAT4 at lower concentrations (10(-6) to 10(-9) M), and had no effect at higher concentrations (10(-4) and 10(-5) M) after the cells were stimulated with PMA + ionomycin. Histamine did not affect IL-12-induced phosphorylation of STAT4. To characterize the histamine receptor subtypes involved in the up-regulation of STAT4 phosphorylation, various H1, H2 and H3/H4 receptor antagonists and/or agonists were employed. H1 receptor agonist (betahistine), but not H2 receptor agonist (amthamine), induced phosphorylation of STAT4. H1 receptor antagonist (pyrilamine) inhibited histamine-mediated phosphorylation of STAT4. However, H2 receptor antagonist (ranitidine) and H3/H4 receptor antagonist (thioperamide) did not alter this effect. Tyrosine kinase inhibitor (tyrphostin) failed to block histamine-mediated phosphorylation of STAT4. These observations suggest that histamine up-regulated the phosphorylation of STAT4 via H1 receptors, and that the Ca2+-PKC pathway, but not the tyrosine kinase pathway, was involved in this effect.     

Expressions of STAT3 and SMAD4 in esophageal squamous cell carcinomas

目的 探讨食管鳞癌组织中信号转导和转录活化因子3(STAT3)、胰腺癌缺失因子4(SMAD4)的表达及其临床意义.方法 采用免疫组化技术检测60例食管鳞癌组织(A组)、28例食管上皮内瘤变组织(B组)和20例食管鳞癌癌旁组织(C组)中STAT3和SMAD4的表达情况.结果 A组STAT3的阳性表达率高于C组(P＜0.05).STAT3的表达与食管鳞癌病理分级呈正相关(P＜0.05).A组和B组SMAD4的阳性表达率低于C组(P＜0.05).SMAD4的表达与淋巴结转移和TNM分期呈负相关(P＜0.05).结论 联合检测STAT3和SMAD4的表达对判断食管鳞癌的预后有一定的指导意义.     

食管鳞癌组织中STAT3与SMAD4的表达

目的探讨食管鳞癌组织中信号转导和转录活化因子3(STAT3)、胰腺癌缺失因子4(SMAD4)的表达及其临床意义。方法采用免疫组化技术检测60例食管鳞癌组织(A组)、28例食管上皮内瘤变组织(B组)和20例食管鳞癌癌旁组织(C组)中STAT3和SMAD4的表达情况。结果A组STAT3的阳性表达率高于C组(P<0.05)。STAT3的表达与食管鳞癌病理分级呈正相关(P<0.05)。A组和B组SMAD4的阳性表达率低于C组(P<0.05)。SMAD4的表达与淋巴结转移和TNM分期呈负相关(P<0.05)。结论联合检测STAT3和SMAD4的表达对判断食管鳞癌的预后有一定的指导意义。     

STAT4 rs7574865单核苷酸多态性与ANCA相关性血管炎的关系

目的 探索信号转导和转录激活因子-4（STAT4）rs7574865单核苷酸多态性与原发性抗中性粒细胞胞浆抗体相关性血管炎（AAV）的关系。方法 纳入145例AAV患者（AAV组）和216例健康体检者（对照组）并收集患者就诊时是否伴发热、蛋白尿、血尿、咯血症状。采用多重PCR结合高通量测序（MPCR-HTS）进行基因分型;比较2组间、各临床症状亚组间rs7574865基因型、等位基因和不同遗传模型下基因型的分布差异;Logistic回归分析评估基因型与临床症状发生风险的关系。结果 2组间STAT4 rs7574865基因型、等位基因、不同遗传模型下基因型分布差异无统计学意义;不同亚组中基因型、等位基因分布差异均无统计学意义。隐性模型下,血尿组TT基因型分布比例低于无血尿组（P<0.05）;Logistic回归分析显示TT基因型与血尿发生风险无关。结论 STAT4 rs7574865单核苷酸多态性与AAV易感性和发热、蛋白尿、血尿、咯血临床症状不相关。     

Association of HLA-DP/DQ and STAT4 polymorphisms with ankylosing spondylitis in Southwest China

Ankylosing spondylitis (AS) is a highly heritable complex inflammatory arthritis disease. Genetic factors are thought to be crucial in the pathogenesis of AS. However, few data are available on the relationship between HLA-DP/DQ and STAT4 polymorphisms and AS susceptibility in the Chinese population. Therefore, we examined HLA-DP/DQ and STAT4 polymorphisms (rs3077, rs9277535, rs7453920 and rs7574865) in a total of 779 subjects, including 400 AS and 379 age- and sex-matched healthy controls in Chinese. No significant difference was observed between AS patients and healthy controls in the allele frequency of rs3077, rs9277535 and rs7574865. However, there was a significant association between the HLA-DQ rs7453920 G/A variant and AS patients, with minor allele A correlated with a reduced risk of AS (allelic frequency, adjusted OR = 0.66, 95% CI = 0.55–0.78, p = 4.0E − 06; dominant model, adjusted OR = 0.75, 95% CI = 0.66–0.85, p = 1.1E − 05). Moreover, the haplotypes block AAA and GGA in the HLA gene significantly correlated with reduced risk of AS. This is the first study demonstrating the significant associations of SNP rs7453920 and the haplotypes in the HLA gene with the risk of AS in Southwest Chinese population. This research sheds new light on the significant relationship between HLA polymorphisms and AS.     

STAT6 and JAK1 are essential for IL-4-mediated suppression of prostaglandin production in human follicular dendritic cells: opposing roles of phosphorylated and unphosphorylated STAT6

Prostaglandins (PGs) are emerging as important immune mediators. Since our first report on the expression of prostacyclin synthase in the germinal centers, we have investigated production mechanisms and biological functions of PG using human follicular dendritic cell (FDC)-like cells. In the previous report, we observed that TGF-β enhances PG production, and IL-4 prevents this upregulation. To elucidate the inhibitory mechanism of IL-4, its effects on the key enzyme leading to PG production were analyzed in this study. IL-4 but not IL-10 inhibited TGF-β-induced COX-2 expression at both mRNA and protein levels. Next the early signaling molecules of IL-4 were identified by siRNA technology. IL-4 induced tyrosine phosphorylation of STAT1, 3, and 6, but only JAK1-STAT6 pathway was responsible for the prevention of COX-2 augmentation and PG production. Phosphorylated STAT6 accumulated in the nucleus rapidly upon IL-4 addition, and the complete inhibition of COX-2 upregulation required 24 h of pretreatment with IL-4, implying that newly transcribed molecules mediate the inhibitory signals downstream of STAT6. Interestingly, unphosphorylated STAT6 proteins were constitutively expressed in the nucleus, and depletion of STAT6 impaired background level expression of COX-2 and PGs. Our results highlight the crucial roles of TGF-β and IL-4 in the regulation of PG production, which lead us to suggest that T cells play an important role in FDC production of PGs.