泌乳素对诱发自身免疫性甲状腺炎影响的实验研究

目的临床上可观察到有时自身免疫性甲状腺炎常伴有高泌乳素血症,产后甲状腺炎也有较高的发病率(5%～10%).本实验研究泌乳素在诱发实验性自身免疫性甲状腺炎(experiment autoimmune thyroiditis,EAT)形成中的影响,进一步探讨泌乳素在自身免疫性甲状腺炎发病中的作用机理.方法选用SD大鼠,雌性,10周龄.大鼠随机分为4组,即SD大鼠在实验的全过程用溴隐亭(Br)灌胃(每只大鼠溴隐亭灌胃量为1mg·kg-1·d-1),诱发EAT,高碘(HI)饮食喂养为Br-HI-EAT组;SD大鼠高碘饮食喂养,诱发EAT为HI-EAT组;高碘饮食对照组(HI组);适碘(NI)饮食正常对照组(NI组).适碘饮食是用低碘饲料(碘含量≤3 5μg/kg)加适碘水(含KI为0.3mg/L)喂养,高碘饮食用低碘饲料加高碘水(含KI为30g/L)喂养.EAT的诱发采用同种属不同品系的Wistar大鼠甲状腺球蛋白(Tg)加完全福氏佐剂(completefreunds adijuvant)免疫SD大鼠(Tg每次1mg,每2周1次,共进行3次)诱发EAT的形成,末次免疫的4周后,处死全部大鼠.各组均观察其大鼠甲状腺组织的病理改变和血清中Tg自身抗体、泌乳素水平及脾脏中的白细胞介素-2/白细胞介素-2受体(IL2/IL-2R)mRNA的表达.结果大鼠经溴隐亭干预后,诱发EAT的高碘饮食(Br-HI-EAT)组,其血清中泌乳素水平为(2.32±0.096)μg/L低于诱发EAT的高碘饮食(HI-EAT)组(3.56±0.048)μg/L,P＜0.05.Br-HI-EAT组的发病率和甲状腺组织的炎细胞浸润(8例中有2例为"++",3例为"+",5例为"-")明显的轻于HI-EAT组(7例中2例为"+++",1例为"++",4例为"+");未诱发EAT的高碘饮食正常对照组(HI组)和正常对照组(NI组)的甲状腺组织中,未发现明显的炎细胞浸润;BR-HI-EAT组大鼠脾脏IL-2和IL-2RmRNA的表达分别为0.27±0.027和0.48±0.039(细胞因子/β-actin)也较HI-EAT组(0.60±0.013和0.75±0.018)明显降低(P＜0.01);诱发EAT两组大鼠血清中Tg自身抗体平均水平明显高于未诱发EAT大鼠(P＜0.01).结论大鼠经溴隐亭于预,致血清中泌乳素水平下降,其IL-2/IL-2RmRNA的表达也随之减少,降低IL-2/IL-2R系统对免疫应答的激活作用,其EAT的发病率和甲状腺组织中炎症反应得到缓解,可以认为泌乳素水平的变化也是影响自身免疫性疾病发病机制中的重要因素.     

瘦素对实验性自身免疫性甲状腺炎影响的实验研究

目的研究瘦素在诱发实验性自身免疫性甲状腺炎（EAT）形成中的影响。方法采用C57BL／6J小鼠在外源性瘦素干预下诱发EAT，观察小鼠甲状腺组织病理改变、淋巴细胞增殖水平、脾细胞培养上清中IFN-γ和IL-4水平以及血清中甲状腺过氧化物酶抗体（TPO—Ab）、甲状腺球蛋白抗体（Tg—Ab）和瘦素水平。结果小鼠经外源性瘦素干预后，其血清中瘦素水平高于EAT组和对照组，实验性自身免疫性甲状腺炎的发病率和甲状腺组织中炎细胞浸润程度也较直接诱发EAT组明显增高。诱发EAT两实验组间比较，脾细胞增殖水平未见明显差异，但瘦素干预组脾细胞培养上清中IFN-γ水平较EAT组明显升高（P=0．000），而IL-4水平则低于EAT组（P=0．000）。两实验组小鼠血清中TPO—Ab和Tg—Ab水平均明显高于未诱发EAT对照组，但两组间比较均未见明显差异。结论瘦素的前炎症作用在实验性自身免疫性甲状腺炎的发生、发展中有重要免疫调节作用。     

碘对诱发易感和非易感小鼠自身免疫性甲状腺炎的实验研究

目的：研究典对实验性自身免疫性甲状腺炎（EAT）易和非易感小鼠诱发EAT的影响，进一步探讨自身免疫性甲状腺疾病（AITD）发病的机理。方法：采用EAT易感（TA－1）和非易感（BALB／c）小鼠经甲状腺球蛋白（Tg）免疫，诱发为EAT实验动物模型并给予不同水平的碘饮食，观察小鼠甲状腺组织的病理改变和炎细胞润程度，血清中Tg自身抗体的水平以及低磺和高碘Tg抗原性的不同。结果：（1）2种品系小鼠经Tg诱发，均出现不同水平的Tg自身抗性和甲状腺的炎症反应，血清中Tg自身抗体的出现早于甲状腺的病理改变。（2）TA－1小鼠的甲状腺炎症反应明显高于BALB／c小鼠，高碘组中的TA－1小鼠炎症较适碘组更明显，为弥漫性伴慢性炎症（＋＋＋）。（3）高碘Tg诱发的Tg抗体与高碘Tg抗原的反应明显高于与低碘Tg抗原的反应，低碘Tg诱发的Tg抗体反之，表明二者抗原性有所不同。结论：高碘摄入会使易感小鼠EAT的发病加重，其机理除了体内的遗传因素，可能与过度碘的Tg抗原性增强有关。     

碘对诱发实验性自身免疫性甲状腺炎形成的影响

目的建立诱发型实验性自身免疫性甲状腺炎(EAT)动物模型,研究碘对诱发EAT形成的影响,进一步探讨其可能的作用机制。方法:采用雌性Lewis大鼠诱发EAT并饲以不同碘含量的饮食,观察各组大鼠甲状腺病理改变、血清中Tg自身抗体水平、血清中甲状腺激素水平、脾细胞分泌TNF水平和甲状腺FasmRNA表达水平。结果1.诱发EAT的HI组大鼠炎症范围较广,为较重度炎症。NI组显示为轻度炎症,且为恢复期。LI组显示为滤泡内少量淋巴细胞浸润。2.诱发EAT的HI、NI和LI组大鼠血清中均出现较高水平的Tg自身抗体,3组间无显著性差异。3.诱发EAT的HI组大鼠脾细胞分泌TNE水平高于NI和LI组(P<0.05)。4.诱发EAT的HI、NI和LI组大鼠甲状腺FasmRNA的表达水平均较各自未诱发EAT的大鼠明显升高,其中HI组高于同组的NI和LI组,P<0.01。结论1.高碘饮食可使已诱发EAT的Lewis大鼠甲状腺组织炎症加重且炎症反应持续时间较长,考虑与脾细胞分泌的TNF水平和甲状腺FasmRNA的表达水平较高有关。2.未发现甲状腺组织炎症的程度与自身抗体水平相关。     

催乳素与实验性自身免疫性甲状腺炎关系的研究

目的　观察催乳素（PRL）在实验性自身免疫性甲状腺炎（EAT）发生发展中的作用。方法　观察SD大鼠发生EAT时PRL水平的变化,以及溴隐停对EAT发生的干预作用。结果　发生EAT的大鼠,其血清PRL水平（8.70±0.42）μg/L明显高于对照组（6.46±0.67）μg/L,P＜0.01,且自身抗体水平也明显高于对照组;而以溴隐停干预的大鼠,随着PRL水平的下降（5.30±0.81）μg/L,自身抗体水平也下降,甲状腺病变亦同时减轻。结论　高催乳素血症在EAT发病中起着重要作用,用溴隐停控制这一环节,对疾病的预防治疗是有益的。     

3种诱导自身免疫性甲状腺炎动物模型的效果比较

目的 采用3种诱导方法建立小鼠实验性自身免疫性甲状腺炎(EAT)动物模型，比较其成功性和效果及实验方法的简易程度。为自身免疫性甲状腺疾病发病机理的深入研究和探讨，提供较理想的EAT动物模型。方法 采用①同种不同系的甲状腺球蛋白(Tg)与完全福氏佐剂(Complete freund’s adjuvant，CFA)免疫实验动物。②同种不同系的甲状腺组织移植入实验动物腹腔中。③同种不同系Tg致敏的淋巴细胞被动转移给同系实验动物，分别诱发EAT动物模型，同时设立正常小鼠作为对照组。4周后，观察甲状腺组织病理改变和检测血清中Tg抗体水平等。结果 Tg CFA组小鼠血清中的自身Tg-抗体水平与正常未免疫组比较，高出约5倍之多，其甲状腺组织亦发生明显的炎症反应，淋巴细胞浸润指数≥2为5例，为本实验组例数的50％。其它2种方法诱发小鼠EAT的实验组中，其甲状腺组织未发现明显的炎症反应和炎细胞浸润。结论 根据形成EAT的2个标准即自身Tg-抗体水平和甲状腺组织淋巴细胞的浸润程度，比较3种诱发小鼠EAT实验方法，可以认为采用Tg cFA免疫动物，建立EAT动物模型有较高的成功性和可靠性，且诱导EAT的实验方法较简单，易行。可为自身免疫性甲状腺疾病发病机理的实验研究提供较满意的动物模型。     

雌激素对大鼠实验性自身免疫性甲状腺炎的影响

目的　研究雌激素在诱发实验性自身免疫性甲状腺炎 (EAT)形成中的影响。方法　 10周龄Wistar大鼠进行双侧卵巢切除术后采用同种属SD大鼠甲状腺球蛋白 (Tg)免疫诱发EAT。病理切片HE染色观察甲状腺炎症反应。采用放免方法测定血清中E2 、PRL水平。间接酶联免疫吸附实验 (ELISA )测定血清Tg抗体 (TgAb)水平。逆转录 聚合酶链反应 (RT PCR)检测甲状腺组织中γ 干扰素 (IFN γ)、白细胞介素 10 (IL 10 )mRNA的表达水平。结果　大鼠切除双侧卵巢后 ,其血清中雌二醇 ( 17.60± 1.0 2 )pmol/L和PRL( 2 .45± 0 .15 ) μg/L水平明显低于未切除卵巢组〔分别为 ( 5 8.5 6± 6.99) pmol/L和 ( 3 .2 2± 0 .17) μg/L〕(均P <0 .0 1) ,实验性自身免疫性甲状腺炎的发病率和甲状腺组织中炎细胞浸润程度也均较未切除卵巢直接诱发EAT组明显减轻 ,甲状腺组织中IFN γmRNA的表达水平 ( 0 .87± 0 .0 3 )较未切除卵巢直接诱发EAT组 ( 0 .99± 0 .0 0 )明显下降 (P <0 .0 5 ) ,而IL 10mRNA的表达水平 ( 0 .98± 0 .0 1)则高于未切除卵巢直接诱发EAT组 ( 0 .83± 0 .0 5 )。诱发为EAT的两组大鼠血清TGAb平均水平 ( 1.3 9± 0 .0 3 )明显高于非诱发EAT对照组的平均水平 ( 0 .18± 0 .0 0 )(P <0 .0 1)。结论　雌激素水平低下可     

碘对易感性不同大鼠诱发自身免疫性甲状腺炎的影响

目的研究碘对实验性自身免疫性甲状腺炎(EAT)易感性不同大鼠诱发EAT的影响,进一步探讨碘和易感性在自身免疫性甲状腺炎(AT)发病中的影响。方法采用猪甲状腺球蛋白(Tg)对不同碘摄入量的Lewis大鼠(EAT易感品系)和Wistar大鼠(EAT非易感品系)进行免疫,诱发EAT形成。观察大鼠甲状腺组织的病理改变和炎细胞浸润程度,用放射免疫分析法检测血清中Tg鄄Ab、TPO(甲状腺过氧化物酶)鄄Ab和甲状腺激素(TT3、TT4)水平。结果①诱发EAT的各组大鼠甲状腺组织均有不同程度的炎细胞浸润,不同碘水平的Lewis大鼠诱发EAT的甲状腺炎症反应均重于诱发EAT的Wistar大鼠,且甲状腺的炎症反应随碘摄入量的增加而加重。②诱发EAT的Lewis大鼠TPO鄄Ab水平明显高于Wistar大鼠各EAT组(P<0.01)。Tg鄄Ab和TPO鄄Ab水平随着碘摄入量的增加亦有升高的趋势。③100倍碘水平的各组T4水平下降(P<0.05),各组的血清T3水平比较差异无统计学意义。结论①EAT易感的Lewis大鼠诱发EAT的甲状腺炎症反应和血清中TPO鄄Ab水平均明显高于EAT非易感的Wistar大鼠。②碘摄入量的增加明显加重EAT的炎症反应,在EAT易感的Lewis大鼠中表现更为明显。③诱发EAT大鼠的甲状腺组织炎症程度和TPO鄄Ab水平相一致。因此,可认为TPO鄄Ab与EAT的发生、发展及甲状腺病变程度的关系更     

雌激素对诱发自身免疫性甲状腺炎影响的实验研究

目的自身免疫性甲状腺疾病(autoimmune thyroid disease,AITD)的发病,女性多于男性,育龄期女性对AITD的易感性更高于男性,存在明显的性别差异.本文研究女性激素中的雌激素对诱发实验性自身免疫性甲状腺炎(experiment autoimmune thyroiditis,EAT)形成的影响,探讨雌激素在AITD发病机制中的作用.方法选用Wistar大鼠,雌性,10周龄.大鼠随机分为4组①切除大鼠的双侧全部卵巢(OVX),高碘(HI)饮食喂养,诱发EAT为OVX-HI-EAT组;③高碘饮食喂养,诱发EAT为HI-EAT组;③高碘饮食对照组(HI组);④适碘饮食正常对照组(NI组).适碘饮食是用低碘饲料(碘含量≤35μg/kg)加适碘水(含KI为0.3 mg/L)喂养,高碘饮食用低碘饲料加高碘水(含KI为15mg/l)喂养,EAT的诱发采用同种属不同品系的SD大鼠甲状腺球蛋白(Tg)加完全福氏佐剂(complete freund's adjuvant)免疫Wistar大鼠(Tg每次1 mg,每周2次,共3次),诱发EAT的形成,末次免疫的4周后,处死全部大鼠.各组均检测血清中雌二醇(Eα-17β)和泌乳素(PRL)水平,观察甲状腺组织炎细胞浸润程度和炎症反应,测定血清Tg抗体水平.并用逆转录-聚合酶链反应(RT-PCR)检测甲状腺组织中干扰素-γ(IFN-γ)和白细胞介素-10(IL-10)mRNA的表达水平.结果去卵巢高碘饮食诱发EAT(OVX-HI-EAT)组大鼠血清中E2-17β(17.60±1.03)pmol/L和PRL(2.45±0.15)μg/L水平均明显低于未切除卵巢的高碘饮食诱发EAT(HI-EAT)组、高碘饮食(HI)组和适碘饮食对照(NI)组,P＜0.01.OVX-HI EAT组诱发EAT的发病率和甲状腺的炎细胞浸润程度(10例中,甲状腺炎细胞浸润1例为"++",5例为"+")明显低于HIEAT组(8例中甲状腺炎细胞浸润5例为"++",2例为"+").甲状腺组织中Th1细胞因子IFN-γmRNA的表达水平,OVX-HI-EAT组为0.87±0.03(细胞因子/β-actin)低于HI-EAT组(0.99±0.002),P＜0.05.OVX-HI-EAT组Th2细胞因子与IL-10mRNA的表达水平为0.98±0.009(细胞因子/β-actin)高于HI-EAT组(0.83±0.05),P＜0.05.诱发EAT大鼠血清中Tg自身抗体的水平明显高于未诱发EAT组大鼠.结论雌激素水平低下可减少或缓解EAT的发病率和甲状腺的炎症反应,高水平雌激素是构成女性对AITD易感的重要因素,雌激素水平低下致Th1细胞偏于Th2细胞可能是EAT发病率和甲状腺炎症反应得以缓解的主要机制.     

硒对大鼠自身免疫性甲状腺炎的影响

目的研究硒对实验性自身免疫性甲状腺炎(EAT)大鼠甲状腺组织结构和自身抗体水平的影响。方法Wistar大鼠随机分为4组:对照组、EAT组、硒预防EAT组、硒治疗EAT组。除对照组外的各实验组均饮用0.05%的碘化钠水溶液,EAT组、硒预防EAT组和硒治疗EAT组在高碘的基础上皮下注射甲状腺球蛋白,建立自身EAT大鼠模型。硒预防EAT组和硒治疗EAT组在不同的时间灌胃给予亚硒酸钠,观察大鼠甲状腺组织变化和自身抗体水平。结果硒干预后甲状腺淋巴细胞浸润明显减少,滤泡破坏程度减轻。EAT组TGAb、TMAb为(62.07±10.01)、(51.50±5.91),硒预防EAT组的TGAb、TMAb为(40.28±6.91)、(35.02±3.67),低于EAT组(P<0.05),硒治疗EAT组的TGAb、TMAb为(42.80±13.90)、(35.00±5.64),低于EAT组(P<0.05),硒预防EAT组和硒治疗EAT组之间差异无统计学意义(P>0.05)。结论高碘的基础上免疫动物可造成大鼠甲状腺组织的破坏,而硒制剂使EAT大鼠病理改变明显减轻,并且在一定程度上预防和抑制其自身抗体水平的升高。     

Activated protein C,an anticoagulant polypeptide,ameliorates severe acute pancreatitis via regulation of mitogen-activated protein kinases

Background Our aim was to investigate the changes of mitogen-activated protein kinases (MAPKs) by activated protein C (APC) treatment in rats with severe acute pancreatitis (SAP), and relate them to changes in SAP severity, thus providing evidence for developing clinical therapies. Methods Sprague-Dawley rats were given an intravenous injection of saline (SAP group), APC (50 μg/kg or 10 μg/kg), or CNI1493 just before SAP induction. One group of rats underwent a sham operation (control group). Experimental samples were harvested 16 h after SAP induction. The gene expression of pancreatic MAPKs was evaluated by cDNA microarrays. The mRNA and protein/phosphorylated protein levels of p38 MAPK, extracellular signal-regulated protein kinase (ERK) 1/2, and c-Jun N-terminal kinase (JNK) and the protein levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined in pancreatic tissue. The severity of disease was evaluated by pancreatic histology, the pancreatic wet/dry weight ratio, and the serum amylase level. Results In rats treated with APC (50 μg/kg) or CNI1493, the severity of pancreatitis and expression of pancreatic TNF-α and IL-1β proteins were attenuated by the decreased expression and activity of p38 MAPK and JNK (vs. the SAP group, P < 0.01). The expression and activity of ERK1/2 were increased in APC-treated rats, especially in the group treated with APC 50 μg/kg (vs. the SAP or CNI1493-treated group, P < 0.01, respectively). Conclusions Inhibition of expression of pancreatic p38 MAPK and JNK and upregulation of ERK1/2 expression by APC treatment may protect against pancreatic injury, thus ameliorating severity of the disease.     

Blockade of the interleukin‐21/interleukin‐21 receptor pathway ameliorates disease in animal models of rheumatoid arthritis

Objective

Interleukin‐21 (IL‐21) is a T cell–derived cytokine that modulates T cell, B cell, and natural killer cell responses. In this study, the effects of blocking IL‐21 were examined in 2 rodent models of rheumatoid arthritis (RA) to determine whether IL‐21 contributes to their pathologic processes.

Methods

DBA/1 mice were immunized with bovine type II collagen and then treated with murine IL‐21 receptor Fc fusion protein (IL‐21R.Fc), which was initiated after the onset of arthritis symptoms in 10% of the cohort. The mice were assessed 3 times per week for signs of disease, including histologic features as well as serum cytokine, Ig, and cytokine messenger RNA (mRNA) levels in the paws. In a separate experiment, Lewis rats were immunized with Freund's complete adjuvant followed by administration of IL‐21R.Fc at the peak of inflammation in the joints. Rats were assessed daily for histologic features and for scoring of arthritis severity. In addition, the effects of IL‐21R.Fc on the production of interferon‐γ (IFNγ) by T cells were examined.

Results

Treatment of DBA/1 mice with IL‐21R.Fc reduced the clinical and histologic signs of collagen‐induced arthritis. Nonspecific IgG1 levels were decreased in response to treatment. The levels of IL‐6 mRNA in the paws and the serum IL‐6 levels were decreased after treatment with IL‐21R.Fc. IFNγ mRNA levels were increased in the paws, and the addition of IL‐21R.Fc to collagen‐activated lymph node cultures enhanced the levels of IFNγ. Collagen‐specific spleen cell responses in IL‐21R.Fc–treated mice were observed as reduced levels of IFNγ and increased levels of IL‐6. Treatment of Lewis rats with IL‐21R.Fc after induction of adjuvant‐induced arthritis resulted in reversal of disease signs and improvements in histologic parameters.

Conclusion

These findings demonstrate a pathogenic role for IL‐21 in animal models of RA, and support consideration of IL‐21 as a therapeutic target in human RA.

     

碘对小鼠产后甲状腺炎影响的实验研究

目的 探讨碘对小鼠产后甲状腺炎(postpartum thyroiditis,PPT)发生,发展的影响.方法 44只8周龄雌性C57BL/6J小鼠均饲以低碘饲料(含碘量≤35 μg/kg),按体质量随机分成4组:非妊娠实验性自身免疫性甲状腺炎(EAT)组 8只,采用猪甲状腺球蛋白加完全弗氏佐剂复制EAT模型,最终存活6只,NI-PPT组(正常碘),10HI-PPT组(10倍碘),50HI-PPT组(50倍碘)各12只,复制EAT模型(方法 同非妊娠EAT组)后,与性成熟雄鼠交配,分别有7,6,6只小鼠受孕.4 组鼠分别饮用含KI为0.3,0.3,3.0,15.0 mg/L的碘水.妊娠母鼠生产4周后,观察4组小鼠甲状腺组织病理改变,血清中甲状腺球蛋白抗体(Tg-Ab)和甲状腺过氧化物酶抗体(TPO-Ab)水平,血清中甲状腺激素(TT3,TT4)水平以及脾脏中IFN-γ,和IL-4 mRNA的表达水平.结果 甲状腺组织病理检查示甲状腺内炎细胞浸润,上皮细胞扁平,滤泡萎缩或破坏.非妊娠EAT组,NI-PPT组,10HI-PVT组小鼠的炎细胞浸润程度均低于50HI-PPT组,组间两两比较,差异有统计学意义(P＜0.05).非妊娠EAT组,NI-PPT组,10HI-PPT组和50HI-PPT组的血清TPO-Ab水平分别为(14.32±8.85)%,(64.45±10.52)%,(38.46±5.57)%,(90.09±9.98)%,任意两组组间比较,差异均有统计学意义(P＜0.05),血清Tg-Ab水平分别为(33.74±3.71)%,(29.65±2.06)%,(37.21±3.87)%,(33.87±4.17)%,组间比较差异无统计学意义(F＝0.484,P＞0.05),血清TT3水平分别为(2.47±0.69)%,(1.57±0.25)%,(1.60±0.28)%,(1.82±0.75)%,组间比较差异无统计学意义(F＝1.596,P＞0.05),非妊娠EAT组,NI-PPT组和10HI-PPT组血清中TT4水平[(99.87±5.97)%,(89.13±7.64)%,(91.05±5.82)%]与50HI-PPT组[(66.68±5.47)%]比较,差异均有统计学意义(P＜0.05).在非妊娠EAT组,NI-PPT组,10HI-PPT组和50HI-PPT组,小鼠脾脏IFN-γmRNA表达水平分别为1.02±0.10,1.37±0.10,1.39±0.12,1.68±0.06,除NI-PPT组和10HI-PVF组的组间比较之外,其他的任意两组组间比较.差异均有统计学意义(P＜0.05),IL-4 mRNA表达水平分别为0.24±0.05,0.35±0.05,0.49±0.04,0.53±0.06.10HI-PPT组,50HI-PPT组与非妊娠EAT组和NI-PPT组比较,差异均有统计学意义(P＜0.05).结论 妊娠及产后易诱发PPT,增加适量碘的摄入量,可降低PPT的炎症反应,而补碘过量也是诱发或增加产后甲状腺炎发生的重要因素.因此,妊娠及产后补碘应遵循合理和科学的原则.     

Implication of interleukin 18 in production of matrix metalloproteinases in articular chondrocytes in arthritis: direct effect on chondrocytes may not be pivotal

OBJECTIVE: To clarify the effect of interleukin (IL) 18 on cartilage degeneration by studying the profile of IL18 receptor (IL18R) on chondrocytes and the direct effect of IL18 on production of matrix metalloproteinases (MMPs), aggrecanases, and tissue inhibitors of metalloproteinases (TIMPs) in articular chondrocytes. METHODS: Monolayer cultured human articular chondrocytes were isolated from non-arthritic subjects and patients with rheumatoid arthritis or osteoarthritis. Gene expression of IL18, IL18Ralpha, IL18Rbeta, MMPs, and aggrecanases was detected by RT-PCR. Protein levels of IL18Ralpha were analysed by flow cytometry. Protein levels of IL18, MMPs, and TIMPs were measured by ELISA. Aggrecanase-2 mRNA expression was quantitatively analysed by real time RT-PCR. Protein levels of signalling molecules were assayed by western blotting. RESULTS: IL18 mRNA was constitutively expressed in chondrocytes, and was enhanced by IL1beta stimulation. Flow cytometric analysis showed that IL1beta, tumour necrosis factor alpha, and IL18 up regulated IL18Ralpha expression levels. The level of IL18Rbeta mRNA was much lower than that of IL18Ralpha, and was slightly up regulated by IL1beta. In chondrocytes responding to IL18, IL18 (1-100 ng/ml) slightly increased the production of MMP-1, MMP-3, and MMP-13, which was blocked by NF-kappaB inhibitor and p38 mitogen activated protein kinase inhibitor. IL18 up regulated mRNA expression of aggrecanase-2, but not aggrecanase-1. IL18 also slightly stimulated TIMP-1 production?through extracellular signal regulated kinase activation. CONCLUSION: IL18 induces production of MMPs from chondrocytes in inflammatory arthritis. Although the direct effect of IL18 on chondrocytes may not be pivotal for the induction of cartilage degeneration, IL18 seems to play some part in the degradation of articular cartilage in arthritis.     

黄芪多糖对大鼠缺氧/复氧诱导的心肌细胞自噬及凋亡抑制作用的机制探讨

目的：探究黄芪多糖（APS）通过调控高迁移率族蛋白1/Toll样受体4/核转录因子-κB(HMGB1/TLR4/NF-κB)信号通路对大鼠缺氧/复氧（H/R）诱导的心肌细胞自噬及凋亡的抑制作用。方法：建立H9C2心肌细胞H/R损伤模型并分为4组：对照组、H/R组、APS组和HMGB1抑制剂组。CCK-8法和EdU染色法检测细胞增殖能力;酶联免疫吸附试验检测细胞肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-6含量;透射电镜观察细胞自噬小体的形成;膜联蛋白V-异硫氰酸荧光素/碘化丙啶（AnnexinV-FITC/PI）双染法检测细胞凋亡;实时定量PCR检测细胞HMGB1、TLR4、NF-κB p65 mRNA表达水平;蛋白免疫印迹法检测细胞HMGB1、TLR4、NF-κB p65、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白（Bax）、含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)、P62、微管相关蛋白1A/1B-轻链3(MAP1LC3,缩写为LC3)-Ⅱ蛋白表达水平。结果：与对照组相比,H/R组细胞增殖能力明显减弱,凋亡及自噬水平明显增加,细胞内可见大量自...     

肺炎支原体感染小鼠Th1／Th2平衡状况及Tim-3mRNA的表达

目的通过检测Tim-3mRNA的表达水平及IFN-7、IL-4的含量,初步探讨肺炎支原体感染鼠Th1／Th2细胞的动态变化及Tim～3的免疫调控作用。方法通过滴鼻感染建立肺炎支原体肺炎小鼠模型,观察小鼠的一般状况,分别于感染后3d、5d、7d、14d、21d取材,应用RT—PCR方法检测肺组织和脾脏Tim-3mRNA的表达;采用ELISA法检测肺泡灌洗液中细胞因子IFN～γ、IL-4的含量。结果实验组IFN-γ于3d出现升高（t=2．309,P〈0．05）,5dIFNY开始下降,而IL-4于3d开始升高并一直持续到7d（t-6．436,P〈0．05）,之后逐渐恢复正常,肺组织和脾脏Tim-3mRNA的表达均于5d达到峰值（t=4．727,P〈0．05;t=7．962,P〈0．05）,且肺组织和脾脏Tim-3mRNA表达量与L-4呈正相关（r=0．561,P〈0．05;r=0．462,P〈0．05）,从5d开始与IFN-γ呈负相关（r=0．379,P〈0．05;r=0．431,P〈0．05）。结论小鼠感染肺炎支原体后Th1／Th2失衡,以Th2介导的免疫反应为主;Tim3可能参与了感染后对Th1细胞的负调控。     

SIRT6-shRNA预处理对缺氧/复氧诱导的H9C2心肌细胞损伤的影响及其机制

目的 探讨通过RNA干扰技术抑制H9C2心肌细胞内的SIRT6基因表达对缺氧/复氧(A/R)诱导的细胞损伤的影响和机制。方法 将H9C2心肌细胞随机分为正常对照组(Con组)、A/R组、阴性对照SIRT6-shRNA质粒处理组(NC组)和SIRT6-shRNA质粒处理组(shRNA组)。检测4组H9C2心肌细胞的存活率、凋亡率、Caspase-3活性及SIRT6、核因子(NF)-κBp65、I-κBα表达水平及细胞培养液中白细胞介素(IL)-6和肿瘤坏死因子(TNF)-α水平并进行比较。结果 与Con组相比,A/R组H9C2心肌细胞存活率和细胞浆中I-κBα蛋白表达水平明显降低,凋亡率、细胞SIRT6-mRNA和蛋白、细胞核中NF-κBp65蛋白表达水平、细胞培养液中IL-6和TNF-α水平及Caspase-3活性明显升高（P<0.05）。与A/R组比较,shRNA组H9C2心肌细胞存活率、细胞SIRT6-mRNA和蛋白及细胞浆中I-κBα蛋白表达水平明显降低,细胞凋亡率、细胞核中NF-κBp65蛋白、细胞培养液中IL-6和TNF-α水平及Caspase-3活性明显升高(P<0.05)。结论 抑制H9C2心肌细胞内SIRT6基因表达能促进炎症因子的分泌,激活NF-κB信号通路,诱导细胞凋亡,加重A/R诱导的心肌细胞损伤。     

Sequential expression of protooncogenes during lectin-stimulated mitogenesis of normal human lymphocytes.

The proliferation of non-neoplastic T lymphocytes is regulated, in part, by the coordinated expression of genes encoding T-cell growth factor (interleukin 2, IL2), IL2 receptors (IL2R), and transferrin receptors (TFR). In addition to growth factors and their receptors, protooncogenes may regulate lymphocyte proliferation. We used cloned cDNAs homologous to 21 different protooncogenes to screen for their expression at the mRNA level in human peripheral blood mononuclear cells (PBMC) stimulated with the mitogenic lectin phytohemagglutinin (PHA), and we compared the time course of accumulation of mRNAs for these protooncogenes to that of mRNAs for the IL2, IL2R, TFR, and histone H3 genes. mRNAs for c-abl, c-ets, c-yes, and N-ras were present in unstimulated PBMC. After stimulation of PBMC by PHA, we detected marked increases within 10 min in the levels of mRNA for c-fos and c-myc; within 6 hr for IL2 and IL2R mRNAs; within 14 hr for c-myb, p53, N-ras, and TFR mRNAs; and within 24-36 hr for H3 mRNA. Expression of c-abl, c-ets, and c-yes increased gradually following stimulation with PHA. None of the other protooncogenes tested was expressed in PBMC. Addition of the protein synthesis inhibitor cycloheximide, before the addition of PHA to cultures, abolished the PHA-induced accumulation of mRNAs for c-myb, N-ras, and TFR, but not of mRNAs for c-fos, c-myc, IL2, and IL2R. These data indicate that c-fos, c-myc, IL2, and IL2R belong to a group of genes expressed early, whereas c-myb, N-ras, and TFR belong to a group of genes expressed later in PHA-activated PBMC, and that the products of the c-fos and c-myc protooncogenes are not required for expression of IL2 or IL2R genes. Addition of purified IL2 augmented the expression of the later-expressed genes c-myb, p53, N-ras, and TFR in PHA-stimulated cultures of PBMC, as well as of the early genes c-myc and IL2R, but not of c-fos and IL2, thus suggesting that PHA and IL2 stimulate the expression of overlapping, but nonidentical, sets of genes in PBMC.