哮喘患者外周血T淋巴细胞凋亡及其分子机制

目的　探讨哮喘患者外周血 Ｔ淋巴细胞凋亡率的变化及其分子机制。方法　取１２ 例急性发作期哮喘患者（ 哮喘组） 及１０ 名正常对照者（ 对照组） 外周血淋巴细胞并无菌分离 Ｔ 淋巴细胞,采用电镜和原位末端终止法（ Ｔ Ｕ Ｎ Ｅ Ｌ） 观察抗 Ｃ Ｄ＋３ 单抗（１００ ｍｇ／ Ｌ） 诱导体外培养０ 、２４ 、４８ 、７２ 小时后 Ｔ淋巴细胞凋亡变化,同时采用细胞原位杂交法观察不同培养时间 Ｔ 淋巴细胞 Ｂｃｌ２ ｍ Ｒ Ｎ Ａ 及 Ｂａｘｍ Ｒ Ｎ Ａ表达变化。结果　正常组及哮喘组 Ｔ 淋巴细胞抗 Ｃ Ｄ＋３ 单抗诱导后均出现凋亡典型形态改变,哮喘患者外周血 Ｔ细胞经抗 Ｃ Ｄ＋３ 单抗诱导２４ 小时后凋亡率为（９１９ ±２２５） ％ ,４８ 小时为（１５８９ ±２１８） ％ ,７２ 小时为（２１３７ ±３２４） ％ ;与正常组２４ 小时（１７９ ±２２２） ％ 、４８ 小时（２５２５ ±３５３） ％ 、７２ 小时（３５１４ ±２５３） ％ 比较,差异有显著性（ Ｐ 均＜ ００１） ,哮喘组７２ 小时方接近正常组２４ 小时水平。哮喘组 Ｔ 淋巴细胞凋亡抑制基因 Ｂｃｌ２ ｍ Ｒ Ｎ Ａ 的表达各时相均与正常组比较,差异有显著性（ Ｐ＜ ００１） 。而凋亡促进基因     

乐胃煎逆转胃癌前病变AgNOR及细胞图像分析

目的研究乐胃煎逆转胃癌前病变不完全结肠型肠化和／或中度异型增生的疗效．方法胃镜病理证实为不完全结肠型肠化和／或中度异型增生４６例．治疗组３０例用乐胃煎，对照组１６例用德诺（Ｄｅ＿Ｎｏｌ）．治疗前后胃镜活检胃窦固定部位粘膜标本作ＡｇＮＯＲ染色及细胞图像分析．结果乐胃煎对不完全结肠型肠化及中度异型增生总有效率均高于Ｄｅ＿Ｎｏｌ，分别为７２％比２５％（Ｐ＜００５）和８９５％比４４４％（Ｐ＜００５）．乐胃煎治疗前后，ＡｇＮＯＲ计数分别为７３０±１１６和４８１±１５０（Ｐ＜００１），Ｄｅ＿Ｎｏｌ组为７７３±０９２和７０５±１０２（Ｐ＜００１）．两组治疗前后ＡｇＮＯＲ计数差值均数相比，统计学上也有显著性差异，分别为２５２±１５４和０６９±０４８（Ｐ＜００１）．乐胃煎组中２０例作细胞图像分析，治疗后各参数（长轴、短轴、核浆比、结构异型指数等）均有不同程度的降低，有显著性差异．结论乐胃煎确有较好地逆转胃癌前病变的功效．     

骨关节炎患者血清抗软骨细胞抗体测定及其意义

目的：为了探讨抗软骨细胞自身免疫反应在骨关节炎（ＯＡ）发病中的可能作用。方法：用培养的人软骨细胞制备抗原，采用酶联免疫法（ＥＬＩＳＡ）检测４１例ＯＡ患者血清抗软骨细胞抗体（ＡＣＡ）水平，并与２９例类风湿关节炎患者（ＲＡ），３３例其它关节炎或关节痛患者及２０名正常人对照。结果：ＯＡ和ＲＡ患者血清ＩｇＧＡＣＡ和ＩｇＭＡＣＡ水平均显著高于其它关节炎或关节痛患者和正常人组（Ｐ＜００１）。ＡＣＡ阳性率在ＯＡ组为４３９％，低于ＲＡ组的８２７％（Ｐ＜０００１），高于其它关节炎或关节痛患者的１８２％和正常人组的５０％（Ｐ＞００５）；有膝关节积液者在ＡＣＡ阳性ＯＡ患者占５５６％，在ＡＣＡ阴性ＯＡ患者仅为１７４％（Ｐ＜００１）。结论：针对软骨细胞的自身免疫反应可能参与ＯＡ的病理过程；ＯＡ滑膜炎的发生可能与ＡＣＡ诱发的免疫反应有关。     

柳氮磺胺吡啶对大鼠乙酸性溃疡性结肠炎肠组织中氧自由基的影响

目的探讨柳氮磺胺吡啶（ＳＡＳＰ）治疗大鼠乙酸性溃疡性结肠炎（ＵＣ）时清除氧自由基（ＯＦＲ）的特性．方法ＳＡＳＰ灌胃治疗大鼠乙酸性ＵＣ后，检测肠组织中的超氧化物歧化酶（ＳＯＤ）、丙二醛（ＭＤＡ）含量，评价其炎症指数，并与生理盐水（ＮＳ）治疗对照组比较．结果ＳＡＳＰ组和ＮＳ组ＳＯＤ含量（Ｕ／ｇ）分别为７９９８±３４４１和６３６４±２４５５．ＳＡＳＰ组和ＮＳ组ＭＤＡ含量（ｎｍｏｌ／ｇ）分别为２１５６±２０８、３５２４±４４８．ＮＳ组和ＳＡＳＰ组炎症指数分别为１６５±５１９、６３０±１２５．ＳＡＳＰ组ＳＯＤ含量显著高于ＮＳ组（７９９８±３４４１对６３６４±２４５５，Ｐ＜００１），ＳＡＳＰ组ＭＤＡ含量明显低于ＮＳ组（２１５６±２０８对３５２４±４４８，Ｐ＜００１）．ＮＳ组炎症指数明显高于ＳＡＳＰ组（１６５±５１９对６３０±１２５，Ｐ＜００１）．结论ＳＡＳＰ为氧自由基清除剂，是治疗溃疡性结肠炎的主要机理之一．     

Ⅲ型肠化是异型增生的一种类型

目的：探讨Ⅲ型肠化与异型增生的关系。方法：采用粘液组化技术将胃粘膜肠化标本分为Ⅰ、Ⅱ、Ⅲ型。盲法比较肠化亚型和异型增生的形态学差异；Ａｇ－ＮＯＲｓ技术定量分析肠化亚型和异型增生细胞Ａｇ－ＮＯＲｓ计数差异。结果：８６％（３７／４３）的Ⅲ型肠化符合轻度异型增生诊断标准；Ⅰ型和Ⅱ型肠化无一例符合（Ｐ＜００００１）；Ａｇ－ＮＯＲｓ计数Ⅲ型肠化（４３３±０８６）显著大于Ⅰ型（２９３±０９２）和Ⅱ型（３１３±０６９）（Ｐ＜００１），但与异型增生（４１４±１４５）差异无显著性（Ｐ＞００５）。结论：Ⅲ型肠化是异型增生的一种类型，提示Ⅲ型肠化很可能是一种癌前病变     

原发性肝癌血清性激素及组织性激素受体的研究

目的研究人体性激素水平变化与原发性肝细胞癌的关系．方法应用放免法测定肝癌组（２０例），肝硬变组（１６例）及正常对照组（２０例）血清雌二醇（Ｅ２）及睾酮（ＴＴＴ）含量，并用放免组化法（ＰＡＰ法）检测肝癌组织及肝硬变组织的雌二醇受体（ＥＲ）及睾酮受体（ＡＲ）含量．结果肝癌组血清Ｅ２含量（４４５５±９３１ｎｇ／Ｌ）明显高于正常对照组（７６６ｎｇ／Ｌ±１７０ｎｇ／Ｌ）而低于肝硬变组（６４９６ｎｇ／Ｌ±１７６ｎｇ／Ｌ）（Ｐ＜００１）；前者ＴＴＴ含量（２５３３００ｎｇ／Ｌ±５６５６０ｎｇ／Ｌ）明显低于后二者（４５８５８０ｎｇ／Ｌ±３４９６０ｎｇ／Ｌ）（Ｐ＜００１）．肝癌组织ＥＲ（８０％）较肝硬变时（４４％）明显增加（Ｐ＜００２５），且与血清Ｅ２含量有明显负相关关系（ｒ＝－０８４７３，Ｐ＜０００１）．ＡＲ阳性百分率在两者无明显差别（ｒ＝－０３１３５，Ｐ＞００５）．结论血清ＴＴＴ含量改变及肝组织ＡＲ浓度改变与肝癌无明显关系，而血清Ｅ２含量改变及肝组织ＥＲ浓度改变与肝癌的发生、发展有密切关系．提示原发性肝细胞癌是一雌激素依赖性肿瘤．     

亚硒酸钠对胃粘膜细胞非程序DNA合成、LPO和ras P21表达的影响

目的研究亚硒酸钠对甲基硝基亚硝基胍（ＭＮＮＧ）所致胃粘膜细胞损伤的防护作用．方法观察了亚硒酸钠对ＭＮＮＧ所致胃粘膜细胞非程序ＤＮＡ合成（ＵＤＳ）、脂质过氧化物（ＬＰＯ）和ｒａｓＰ２１表达的影响．结果胃粘膜细胞先用１０μｍｏｌ／Ｌ或１μｍｏｌ／Ｌ亚硒酸钠预处理４ｈ，再给ＭＮＮＧ组细胞的非程序ＤＮＡ合成水平（ｃｐｍ／×１０－６ｍｉｎ－１，１１６６±１５６或１５６６±１８７ｖｓ１８３８±２０５，Ｐ＜００１～００５），脂质过氧化物（２０ｄ，μｍｏｌ／Ｌ，４５±０６或４７±０６ｖｓ７４±０７，Ｐ＜００１）和ｒａｓＰ２１蛋白含量（２０ｄ，Ａ，０６８±００８或０８６±００７ｖｓ１０８±０１１，Ｐ＜００１～００５）均显著低于ＭＮＮＧ组．结论一定剂量亚硒酸钠对ＭＮＮＧ诱导的胃粘膜细胞损伤有防护作用．     

肝癌患者外周血树突状细胞免疫功能低下

目的研究肝癌患者外周血树突状细胞（ＤＣ）表面免疫分子ＨＬＡＤＲ及Ｂ７表达水平及其与ＤＣ免疫功能的相关性．方法以健康成人（ｎ＝１０）作为对照，检测临床确诊中晚期肝癌（ＨＣＣ）患者（ｎ＝１０）外周血ＤＣ表面免疫分子ＨＬＡＤＲ及Ｂ７表达水平及ＤＣ免疫诱导Ｔ淋巴细胞增殖的能力．结果ＨＣＣ患者ＤＣ表面ＨＬＡＤＲ及Ｂ７表达水平（ＶＯＦ）为６７±１６及６１±１１，明显低于对照组（１０７±１４及９６±１２，Ｐ＜００５）．其ＤＣ体外诱导Ｔ细胞增殖能力（ｍｉｎ－１）为３１００±１２０，亦明显低于对照组（６２００±９０，Ｐ＜００１）．结论．ＨＣＣ患者ＤＣ表面ＨＬＡＤＲ及Ｂ７表达水平下降，并与ＤＣ免疫功能低下密切相关．     

幽门螺杆菌感染对胃上皮细胞增殖和凋亡的影响

目的为了探讨幽门螺杆菌感染对胃粘膜上皮细胞动力学的影响。方法应用免疫组织化学和切口末端标记法（ＴＵＮＥＬ），检测了１６例正常胃粘膜者和３１例幽门螺杆菌（Ｈｐ）相关慢性胃炎患者治疗前后胃粘膜上皮细胞增殖细胞核抗原（ＰＣＮＡ）标记指数（ＬＩ％）、细胞凋亡指数（ＡＩ）和表皮生长因子受体（ＥＧＦ－Ｒ）的表达。结果Ｈｐ阳性患者的ＰＣＮＡＬＩ％为１３．９４±１．６４，正常对照组为６．７１±０．９２，差异有非常显著性（Ｐ＜０．０１）；ＥＧＦ－Ｒ表达与ＰＣＮＡＬＩ％呈正相关（ｒ＝０．４４８７，Ｐ＜０．０１）：Ｈｐ阳性患者组的ＡＩ为７．１±１．６，正常对照组为１．３±０．６，差异有非常显著性（Ｐ＜０．０１）；抗Ｈｐ治疗后，２１例Ｈｐ根除者的ＰＣＮＡＬＩ％和细胞ＡＩ分别降至８．２１±１．３２和１．２±０．６，与治疗前相比差异有非常显著性（Ｐ＜０．０１），而１０例Ｈｐ持续阳性者则无明显降低（Ｐ＞０．０５）：ＰＣＮＡＬＩ％、ＥＧＦ－Ｒ表达及细胞ＡＩ与胃粘膜炎症程度无显著相关（Ｐ＞０．０５）。结论上述结果提示，Ｈｐ感Ｈ能引起胃粘膜上皮细胞过度增殖和凋亡。这为Ｈｐ感染胃癌发病中的作用机制提供了一些线索。     

多次小睡潜伏时间试验在诊断嗜睡症的应用

目的多次小睡潜伏时间试验（ＭＳＬＴ）客观评价嗜睡严重程度、治疗效果及鉴别诊断。方法对１２例正常人、１７例发作性睡病及１２例阻塞性睡眠呼吸暂停综合征（ＯＳＡＳ）患者进行ＭＳＬＴ检查。结果正常对照平均ＭＳＬＴ为２０．１±６．７分；快速眼动睡眠（ＲＥＭ）０．１±０．３次。发作性睡病组平均ＭＳＬＴ３．１±１．９分（与正常组比较，Ｐ＜０．０１；与ＯＳＡＳ组比较，Ｐ＜０．０５）；ＲＥＭ睡眠次数３．４±１．４次（与正常组比较，Ｐ＜０．０１；与ＯＳＡＳ组比较，Ｐ＜０．０５）。ＯＳＡＳ组平均ＭＳＬＴ５．７±３．２分，ＲＥＭ睡眠次数１．７±１．４。结论ＭＳＬＴ试验的平均睡眠潜伏时间及ＲＥＭ发生的次数对发作性睡病及ＯＳＡＳ患者的嗜睡严重程度及对发作性睡病的诊断和鉴别诊断具有临床价值     

三氧化二砷对类风湿关节炎滑膜细胞凋亡影响的体外研究

目的 探讨类风湿关节炎(RA)患者滑膜细胞的凋亡异常,以及三氧化二砷(As_2O_3)对体外培养RA的滑膜细胞凋亡的诱导作用.方法 用光镜、电镜、流式细胞仪检测As_2O_3对体外培养的RA滑膜细胞凋亡的诱导作用,As_2O_3,作用于RA滑膜后酶联免疫吸附试验(ELISA)检测凋亡过程中细胞色素C的变化,反转录-聚合酶链反应(RT-PCR)检测Caspase-3.Bel-2 mRNA表达,采用单因素方差分析和LSD-t检验、Dunnet-t检验进行统计学处理.结果 流式细胞仪检测发现不同浓度的As_2O_3作用后,滑膜细胞的凋亡较空白对照组(NC)增加[NC(0.95±0.87)%,RA(0.21±0.12)%,P<0.05],呈一定的剂量依赖性,尤其以80 μmol/L药物浓度最为明显.凋亡早期(6 h)细胞色素C水平升高80 μmol/L组升高明显[NC(0.34±0.27),80 μmol/L组(32.04±1.62),P<0.05];不同浓度的As_2O_3作用后细胞中Caspase-3 mRNA表达显著增强[NC(0.144±0.022),RA(0.323±0.047),(0.824±0.109),(1.213±0.196),P<0.05],Bcl-2的mRNA表达显著减弱[NC(1.08±0.23),RA(0.94±0.15),(0.46±0.08),(0.22±0.06),P<0.05].结论 RA患者滑膜细胞凋亡较健康埘照减少,As_2O_3通过升高细胞色素c及Caspase-3,抑制Bcl-2诱导RA滑膜细胞凋亡.     

Beneficial effect of galectin 9 on rheumatoid arthritis by induction of apoptosis of synovial fibroblasts

OBJECTIVE: To compare the expression of galectin 9 (Gal-9) in synovial tissue (ST) from rheumatoid arthritis (RA) patients and osteoarthritis (OA) patients and to evaluate the effects of Gal-9 on fibroblast-like synoviocytes (FLS) in these patients. METHODS: The expression of Gal-9 in ST and FLS was compared using immunohistochemical techniques. Apoptotic cells in RA and OA ST samples were detected by TUNEL assay. Apoptosis of FLS was analyzed by the sub-G(1) method in vitro. The in vivo suppressive effects of Gal-9 on collagen-induced arthritis (CIA) in a mouse model were also elucidated. RESULTS: The percentage of Gal-9-positive cells in ST samples and the amount of Gal-9 in synovial fluid samples were significantly higher in patients with RA than in patients with OA, suggesting the involvement of Gal-9 in the development of RA. Compared with the 2 wild-type Gal-9 forms, stable Gal-9, a mutant protein resistant to proteolysis, significantly induced apoptosis of FLS from RA patients. In contrast, other galectins, such as Gal-1, Gal-3, and Gal-8, did not induce apoptosis or suppress the proliferation of human RA FLS. Stable Gal-9 preferentially induced apoptosis and suppressed the proliferation of RA FLS in vitro. It also induced apoptosis of cells in RA ST implanted into SCID mice in vivo. In a mouse model of CIA, apoptotic cells were detected in the joints of stable Gal-9-treated mice, but not phosphate buffered saline-treated mice, and suppressed CIA characterized by pannus formation with inflammatory cell infiltration and bone/cartilage destruction. CONCLUSION: Gal-9-induced apoptosis of hyperproliferative RA FLS may play a critical role in the suppression of RA.     

Localization of human glucocorticoid receptor in rheumatoid synovial tissue of the knee joint

OBJECTIVE: This study was conducted to investigate the localization of human glucocorticoid receptors (GCRs) in the knee synovium of patients with rheumatoid arthritis (RA) and to evaluate the correlation between GCR expression and the clinical profiles. METHODS: Twenty synovial specimens from RA knees, six from knees with osteoarthritis (OA), and five from knees with traumatic arthritis (TA) were obtained at surgery. The GCRs were stained immunohistochemically. The immunopositive cells were counted at random in the lining (synoviocytes) and sublining layers (fibroblastic and lymphoid cells). The relationship between the GCR-expressing cells and clinical profiles was analysed statistically. RESULTS: GCRs were expressed in the nuclei of synoviocytes and the fibroblastic and lymphoid cells in the sublining layer. The GCR-positivity rate of synoviocytes was 67.1+/-18.4% in RA, 58.7+/-13.5% in OA, and 49.4+/-19.7% in TA, differences between the three groups being statistically insignificant. There was a significant difference in the GCR-positivity rate of sublining fibroblastic cells (p = 0.029), but not synoviocytes or sublining lymphoid cells, from RA patients treated with and without prednisolone, while there was no correlation between the rate for synoviocytes and that for sublining fibroblastic cells from RA patients treated with prednisolone. CONCLUSIONS: GCRs are localized not only on inflammatory lymphoid cells but also on synoviocytes, suggesting that glucocorticoids could act directly on these cells. Furthermore, the rate of GCR expression on synoviocytes and sublining lymphoid cells is less suppressed with low-dose prednisolone, regardless of the duration of treatment.     

Down-regulation of FLIP sensitizes rheumatoid synovial fibroblasts to Fas-mediated apoptosis

OBJECTIVE: Hyperplasia of fibroblast-like synoviocytes (FLS) contributes to chronic inflammation and joint destruction in rheumatoid arthritis (RA). FLICE-inhibitory protein (FLIP) is an antiapoptotic protein that might prevent apoptotic elimination of FLS in response to death ligands such as tumor necrosis factor alpha (TNFalpha) or Fas ligand, which are present in RA synovium. Previous studies on FLIP expression by osteoarthritis (OA) and RA FLS have shown variable results, and the specific role of FLIP as an apoptosis inhibitor in these cells remains unclear. We undertook this study to investigate the expression and antiapoptotic function of FLIP in FLS. METHODS: We studied the expression of FLIP by immunohistochemistry and immunoblotting in synovial tissues or cultured FLS from RA and OA patients. FLS apoptosis was induced by an agonistic anti-Fas monoclonal antibody and FLS were then quantified. We studied the effects of cycloheximide (CHX), TNFalpha, and FLIP antisense oligonucleotide on FLIP expression and FLS apoptotic susceptibility. RESULTS: FLIP(L) was the isoform mainly expressed in lining synoviocytes and cultured FLS. Synovial tissues and cultured FLS from OA and RA tissues displayed similar patterns and levels of expression of FLIP. Fas-induced apoptosis was variable in different FLS lines, but differences between OA and RA groups were not detected. TNFalpha induced increases in FLIP(L) and FLIP(S) expression and protected RA FLS from apoptosis, while CHX induced the opposite effects. Down-regulation of FLIP by antisense oligonucleotide strongly sensitized RA FLS to Fas-mediated apoptosis. CONCLUSION: Apoptosis susceptibility and FLIP expression are similar in OA and RA FLS. Down-regulation of FLIP sensitizes RA FLS to Fas-mediated apoptosis and may be a valuable tool for targeting RA FLS hyperplasia.     

Local cell proliferation in rheumatoid synovial tissue: analysis by cyclin expression

The immunohistochemical staining of cyclins was done to evaluate the proliferating cells in synovial tissue of rheumatoid arthritis (RA). Synovial specimens obtained from 18 patients with RA, 12 with osteoarthritis (OA), and 8 with traumatic arthritis (TA) were used for immunostaining of cyclins A and B1 and proliferating cell nuclear antigen (PCNA). The positive cells in lining layer (synoviocytes) and sublining layer (lymphoid and nonlymphoid cells) were counted. Moreover, the relationship between the frequency of their positive cells and clinical data of RA patients was analyzed statistically. In general, cyclin-A-, cyclin-B1-, and PCNA-positive cells in RA were more frequently observed as compared with those in OA and TA. Significant differences were found between RA and OA or TA in cyclin-A-, cyclin-B1-, and PCNA-positive sublining lymphoid cells, between RA and OA or TA in cyclin-B1- and PCNA-positive sublining nonlymphoid cells, and between RA and OA in cyclin-B1-positive synoviocytes. The ratio of cyclin-A- or cyclin-B1-positive cells per PCNA-positive cells was significantly higher in sublining lymphoid cells in RA than TA and in sublining lymphoid and nonlymphoid cells of RA than OA or TA. Moreover, a better relationship was observed between the frequency of cyclin-A-positive synoviocytes and age and between cyclin-A-positive sublining nonlymphoid cells and duration of the disease in RA patients. Our data demonstrated clearly that synoviocytes, as well as sublining lymphoid and nonlymphoid cells, could divide in situ, and more frequent cell division and a higher ratio of cyclin-A- or cyclin-B1-positive/PCNA-positive sublining cells could occur in RA than OA and TA.     

Study of P53 in peripheral blood and synovial mononuclear cells of rheumatoid arthritis and osteoarthritis patients and its relation to the degree of disease activity

Over expression of P53 has been described in many inflammatory conditions including rheumatoid arthritis (RA) and osteoarthritis (OA) as a protective mechanism to induce apoptosis of synovial cells. Lack of P53 function through mutation in human synoviocytes increases the development of normal synovial fibroblasts into transformed aggressive synovial fibroblasts. P53 levels were determined in supernatant of cultured mononuclear cells (MCs) isolated from peripheral blood (PBMCs) of patients with RA (n = 10) and OA (n = 10) as well as 10 normal healthy controls (C). P53 levels were also determined in supernatants of MCs isolated from synovial fluid (SFMCs) of RA and OA patients. Results of this work revealed that P53 level was significantly higher in PBMCs supernatant of RA group than those of both (C) and (OA) groups (P = 0.022). P53 level was non-significantly higher in SFMCs supernatant of RA than OA group. Significantly higher levels of P53 was detected in SFMCs culture supernatant than that of PBMCs within each RA (P = 0.003) and OA (P = 0.001) group. Results also showed a significantly positive correlation between P53 levels (in both PBMCs and SFMCs) and the disease activity score (DAS) in RA group (P = 0.01, P = 0.02 respectively) while insignificantly positive correlations between P53 level (in both PBMCs and SFMCs) and radiological grading of OA group were obtained. These results indicate that mutations and consequent dysfunction of P53 gene may result in chronic inflammation and hyperplasia in RA patients. In conclusion, gene therapy targeting P53-dependent pathway could be a promising therapy for RA and OA diseases.     

Apoptosis of rheumatoid synovial cells by statins through the blocking of protein geranylgeranylation: a potential therapeutic approach to rheumatoid arthritis

OBJECTIVE: To determine whether statins induce apoptosis in rheumatoid arthritis (RA) synoviocytes. METHODS: The effects of lipophilic and hydrophilic statins (fluvastatin and pravastatin, respectively) on the apoptosis of cultured RA synoviocytes were examined in vitro. Apoptosis was analyzed by flow cytometry after staining with JC-1 (to measure the mitochondrial transmembrane potential), active caspase 3, annexin V, and propidium iodide. Add-back experiments were conducted to determine which downstream products of the mevalonate pathway could suppress apoptosis. Modulation of various signaling pathways induced by statins, including protein prenylation, was also investigated. RESULTS: Fluvastatin, but not pravastatin, induced apoptosis in RA synoviocytes in a concentration-dependent (1-10 microM) and time-dependent (48-96 hours) manner. Another lipophilic statin, pitavastatin, displayed almost the same effects as fluvastatin. In sharp contrast, lipophilic statins did not significantly increase apoptosis in synoviocytes from patients with osteoarthropathy. Apoptosis induced by fluvastatin was mitochondrial- and caspase 3-dependent and was abrogated by mevalonate and geranylgeranyl pyrophosphate, but not by farnesyl pyrophosphate. In addition, the geranylgeranyl transferase inhibitor GGTI-298 mimicked the effect of fluvastatin on RA synoviocytes. Treatment of RA synoviocytes with the RhoA kinase inhibitor Y-27632 caused apoptosis. Fluvastatin decreased the amount of RhoA protein in the membrane fraction, but increased the amount in the cytosolic fraction. CONCLUSION: Fluvastatin induced apoptosis in RA synoviocytes through a mitochondrial- and caspase 3-dependent pathway and by the blockage of mevalonate pathways, particularly through the inhibition of protein geranylgeranylation and RhoA/RhoA kinase pathways. These findings suggest that lipophilic statins have potential as novel therapeutic agents for RA.     

C-myc反义寡脱氧核苷酸抑制类风湿滑膜细胞增生诱导滑膜细胞凋亡

目的 研究硫代磷酸化修饰的c—myc反义寡脱氧核苷酸(antisens phosphorothioate oligodeoxynucleotides，ASP-ODN)在血清存在条件下对培养的人类风湿关节炎(rheumatoid arthritis，RA)滑膜B型细胞增生的影响及诱导凋亡的作用。方法 培养RA滑膜B型细胞，合成c—myc AS P-ODN，以阳离子脂质体。lipofectin为载体转染滑膜B型细胞，用显微镜观察和四唑盐(MTT)法检测对细胞增生的抑制作用，用荧光染色、荧光显微镜、流式细胞仪检测诱导细胞凋亡和c—myc蛋白表达的变化。结果c—myc AS P-ODN呈序列特异性抑制滑膜B型细胞增生并下调c—myc蛋白表达，诱导滑膜B型细胞凋亡。结论 c—myc AS P-ODN可能对RA的滑膜增生性炎症具有潜在的治疗作用，c—myc原癌基因可能成为反义核酸技术治疗RA的靶基因。