温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA,TNF-α及肝细胞凋亡的影响

目的:研究Toll样受体4(TLR4) mRNA及其下游炎性因子肿瘤坏死因子-α(TNF-α)与肝衰竭肠源性内毒素血症(IETM)大鼠肝细胞凋亡的关系,并探索温阳解毒化瘀颗粒对内毒素介导的肝细胞凋亡的调控机制。方法:SPF级雄性SD大鼠85只,随机分为正常组,模型组,TLR4单克隆抗体组和温阳解毒化瘀颗粒组(8.925 g·kg^-1)。采用D-半乳糖胺(D-Gal)腹腔注射建立肝衰竭IETM模型,TLR4单克隆抗体组和温阳解毒化瘀颗粒组在造模前5 d给予温阳解毒化瘀颗粒溶液灌胃,正常组、模型组以等容积蒸馏水代替,直至处死。各组分别在24,48,72 h随机处死大鼠并采集标本。检测24,48,72 h各组时间点血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST)水平,苏木素-伊红(HE)染色观察肝组织病理变化,实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织TLR4 mRNA表达,酶联免疫吸附测定法(ELISA)检测肝组织TNF-α表达,流式细胞仪检测肝细胞凋亡率。结果:与正常组比较,模型组ALT,AST升高,肝组织病理损伤程度明显加重,TLR4mRNA,TNF-α表达均增高(P<0.05,P<0.01),肝细胞凋亡率明显上升(P<0.05,P<0.01);与模型组比较,温阳解毒化瘀颗粒组ALT,AST显著降低(P<0.01),肝组织病理损伤程度明显减轻(P<0.05),TLR4 mRNA,TNF-α表达显著下降(P<0.01),肝细胞凋亡率亦显著降低(P<0.01)。结论:TLR4 mRNA,TNF-α在肝衰竭时与肝细胞凋亡呈正相关,温阳解毒化瘀颗粒能够改善肝衰竭IETM大鼠肝功能,减轻肝细胞损伤,减少肝细胞凋亡,其机制可能与其下调肝脏TLR4 mRNA表达,抑制TNF-α释放,降低肝细胞凋亡率有关。     

Kombucha tea ameliorates experimental autoimmune encephalomyelitis in mouse model of multiple sclerosis

Experimental autoimmune encephalomyelitis (EAE) is a mouse model for multiple sclerosis (MS), in which an inflammatory demyelination and axonal damage occurs. Kombucha tea is a fermented beverage made from kombucha mushroom, brewed tea, and sugar. In recent years kombucha tea has attracted interest due to its pharmacological properties like antioxidant effects. The aim of the present research was to test the therapeutic effect of kombucha tea in EAE. We induced EAE model in 18 female C57BL/6 mice by inoculation of myelin oligodendrocyte glycoprotein-35-55 (MOG_35-55) in complete Freund’s adjuvant emulsion. Then, in order to ameliorate EAE symptoms, we used kombucha tea. During the course of study clinical evaluation was assessed, and on the day 21 post-immunization, for evaluation of nitric oxide (NO), total antioxidants capacity and tumor necrosis factor-alpha (TNF-α), blood samples were taken from the heart of mice. The mice were sacrificed and brains and cerebellums of mice were removed for histological analysis. Our findings demonstrated that kombucha tea had beneficial effects on EAE by lower incidence, attenuation in the severity, and also a delay in the onset of disease. Histological analysis showed that inflammatory criteria including the number of infiltrated immune cells and plaques as well as demyelination in kombucha tea dosed mice were significantly lower than the control group. Also, in comparison with control mice, the serum levels of NO and TNF-α in kombucha tea-treated mice were significantly decreased. Kombucha tea with its potential therapeutic effects and immunomodulatory properties might be proposed, after additional necessary tests and trials, for treatment of MS.     

子宫内膜异位症患者血清TNF-α和TNF-β的测定

目的:了解子宫内膜异位症（简称内异症）患者血清中肿瘤坏死因子-α(TNF-α)和肿瘤坏死因子-β（TNF-β）的水平以及在腹腔镜保守性手术治疗前后的变化。方法:采用酶联免疫吸附法检测82例内异症患者（内异症组）和68例非内异症妇女（对照组）血清中TNF-α和TNF-β的含量及49例手术前后两者水平的变化。结果：内异症组血清TNF-α和TNF-β含量均显著高于对照组（Ｐ<0.01），且二者表达量随病情加重有上升趋势（Ｐ<0.05）。手术后Ⅲ～Ⅳ期患者血清中的TNF-α和Ⅰ～Ⅳ期患者血清TNF-β的含量随着内异灶的清除逐渐下降。结论:检测患者血清中TNF-α和TNF-β的含量，对术后随访、监测及手术效果的评价具有重要意义。     

Anti-inflammatory effect of erythropoietin pretreatment on cardiomyocytes with hypoxia/reoxygenation injury and the possible mechanism

Objective： To investigate the anti-inflammatory effect of erythropoietin （EPO） pretreatment on cardiomyocytes exposed to hypoxialreoxygenation injury （H/R） and explore the possible mechanism.
Methods： The cultured neonatal rats＇ ventricular cardiomyocytes were divided randomly into 4 groups, control group （C group）, EPO pretreatment group （E group）, EPO and pyrrolidine dithiocarbamate （PDTC） pretreatment group （EP group） and PDTC pretreatment group （P group）. After 24 hours＇ pretreatment, the cardiomyocytes were exposed to H/R. After pretreatment and H/R, the expression of tumor necrosis factor- α （TNF- α ） gene in all the groups was detected by RT-PCR and Western blot. The nuclear factor- κ B （NF- κB） activity was detected by electrophoretic mobility shift assay （EMSA） and the inhibitor- κB α （Ⅰ- κB α） protein level was detected by Western blot.
Results： The decrement of Ⅰ- κB a protein and the increasing NF- KB activity were found in cardiomyocytes pretreated with EPO before H/R compared to other groups （t=3.321, 4.183, P〈0.01）. However, after H/R, NF- κB activity and expression of TNF- α gene were significantly reduced, Ⅰ- κB a protein expression was increased in cardiomyocytes of E group compared to other groups （t=-3.425, 3.687, 3.454, P〈0.01）. All theses changes caused by EPO pretreatment were eliminated by the intervention of PDTC （an antagonist to NF- κB） during pretreatment.
Conclusions： EPO pretreatment can inhibit the activation of NF- κB and upregulation of TNF- α gene in cardiomyocytes exposed to H/R through a negative feedback of NF- κB signaling pathway, and thus produces the anti-inflammatory effect. This might be one of the ways EPO produces the anti-inflammatory effect.     

精神分裂症患者致炎性细胞因子和酪氨酸羟化酶mRNA表达水平的研究

目的探讨精神分裂症的外周神经免疫机制及其与临床症状的关系。方法检测精神分裂症患者致炎性细胞因子白介素-1β（IL-1β）、肿瘤坏死因子-α（TNF—α）以及酪氨酸羟化酶（TH）的mRNA表达水平，采用逆转录-聚合酶链反应及半定量检测技术，分别检测39例精神分裂症患者（患者组）、25例同胞（同胞组）及30名正常对照（对照组）外周血单个核细胞（PBMC）IL-1β、TNF-α及TH基因表达水平，同时应用阳性和阴性症状量表（PANSS）评定精神分裂症患者临床症状。结果患者组、同胞组及对照组IL-1β的mRNA表达水平分别为1．52±1．01、1．52±1．09和0．74±0．38；TNF—α的mRNA表达水平分别为1．18±0．99、1．01±0．87和0．70±0．29；TH的mRNA表达水平分别为0．55±0．33、0．61±0．32和0．28±0．20。患者组和同胞组的IL-1β、TNF—α、TH的mRNA表达水平均明显高于对照组（P〈0．05或P〈0．01）。患者组IL-1β（r=0．420）、TNF—α（r=0．430）的mRNA表达水平与PANSS的-般病理症状分呈正相关（P〈0．01）。同胞组与对照组合并统计，IL-1β与TNF-α的mRNA表达水平呈正相关（r=0．847，P〈0．01）；IL-1β与TH的mRNA表达水平呈正相关（r=0．666，P〈0．01）。患者组仅IL-1β与TNF—α的mRNA表达水平呈正相关（r=0．942，P〈0．01）。结论精神分裂症患者PBMC细胞TH、IL-1β和TNF—α的mRNA表达水平高于正常，且与精神分裂症的-般病理症状显著相关。     

Are cytokines possible mediators of cancer cachexia?

The possible role of cytokines in the development of cancer cachexia was reviewed from the literature. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, IL-6, interferon (IFN)-gamma and leukemia inhibitory factor (LIF) can elicit many but not all host changes seen in cancer cachexia, including loss of appetite, loss of body weight, and the induction of acute-phase protein synthesis. However, these cytokines are not always demonstrated in the circulation of the cancer patients. The inability to detect circulating cytokines may be due to their low rate of production, their short half-life and rapid clearance from plasma, or their mode of action (autocrine or paracrine). Different cytokines are induced to stimulate the same response. This is very different from hormonal regulation, where a hormone acts on a cell directly through a specific receptor without depending on other mediators. Specific antibodies including anti-IFN-gamma, anti-TNF and anti-IL-6 antibodies, as well as the cyclooxygenase inhibitor indomethacin, have been used to reverse cancer cachexia. Overlapping physiologic activities make it unlikely that a single substance is the sole cause of cancer cachexia. It is hoped that further investigation on other cytokines and their possible relationships with hormones will help to clarify the mechanisms of cancer cachexia in the near future.This work was supported by a grant from the Japan-Sweden Foundation in 1991.     

Monocyte cytokine secretion induced by chemically-defined derivatives of Klebsiella pneumoniae.

The capacity of a K. pneumoniae membrane proteoglycan (Kp-MPG) and four of its chemically defined derivatives to activate human monocytes was studied by measuring immunoreactive IL-1 beta, IL-6 and tumour necrosis factor-alpha (TNF-alpha) in culture supernatants. Monocyte culture supernatants were also tested for their comitogenic activity on concanavalin A-stimulated thymocytes and for their cytotoxic activity on the mouse fibroblastic L929 cell line. The four Kp-MPG derivatives were: (i) an acylpoly(1-3)galactoside (APG); (ii) an APG preparation submitted to acid hydrolysis which removed all fatty acids but left intact the galactose chain of APG (GC-APG); (iii) a preparation obtained by mild alkaline hydrolysis, containing additional ester-linked C14 and C16 fatty acids bound to the APG molecule (EFA-APG); and (iv) a polymer of the latter compound (APG pol). Kp-MPG induced the synthesis of IL-1 beta, IL-6 and TNF-alpha with dose-responses and kinetics similar to those of Salmonella minnesota lipopolysaccharide (Sm-Re-LPS). APG pol and EFA-APG induced the secretion of the three cytokines with lower potency than Kp-MPG or Sm-Re-LPS. APG did not trigger any detectable cytokine production and GC-APG induced only borderline and inconsistent responses. Our data demonstrate the critical role of ester-linked C14 and C16 fatty acids in the triggering of monocyte response to Kp-MPG derivatives.     

A tumour necrosis factor-alpha (TNF-α) promoter polymorphism is associated with chronic hepatitis B infection

Cytokines such as TNF-α and interferon gamma (IFN-γ) are important for the elimination of infected hepatocytes during acute hepatitis B virus (HBV) infection. Two G versus A transitions in the TNF-α promoter region at positions ?308 and ?238 possibly influence TNF-α expression. We investigated these TNF-α polymorphisms in 71 patients with chronic HBV infection, in 32 subjects that had spontaneously recovered from acute HBV infection, and in 99 healthy controls. The ?238 A promoter variant was present in 18 (25%) of 71 patients with chronic HBV infection compared with two (6%) of 32 subjects with acute infection (P < 0.04), and seven (7%) of 99 controls (P < 0.003). By contrast, the prevalence of the variant at position ?308 was similar in all investigated groups. The observed differences could not be explained by linkage disequilibrium to HLA-B or -DRB1* alleles. These findings suggest an association between the TNF-α promoter polymorphism at position ?238 and the development of chronic HBV infection. This promoter variant appears to be linked to defective viral clearance.     

肿瘤坏死因子 α抑制剂对妊娠的影响

 肿瘤坏死因子 α（TNF-α）是类风湿关节炎（RA）发病和维持关节慢性滑膜炎症反应的最重要的致炎性细胞因子之一，它在 T 细胞依赖的炎症性肠病（IBD）发病机制中也起着十分重要的致炎作用。大量临床研究证实，TNF-α抑制剂（TNFAI）可改善 RA 患者关节功能，减少 RA临床活动性，并延缓关节损坏的进展^[1]。各种 TNFAI 也逐渐用于治疗其他风湿性疾病，如银屑病、幼年特发性关节炎、强直性脊柱炎等。某些 TNFAI 被证明对难治性 IBD 有效。目前经美国 FDA 批准上市的 TNFAI 类药物共有 3 种，它们是依那西普（Etanercept，商品名Enbrel），英利西单抗（Infliximab，商品名 Remicade）和阿达木单抗（Adalimumab，商品名Humira）。依那西普是 II 型TNF受体（TNFR-II）与 IgG1-Fc 的融合蛋白，用药方式为皮下注射。英利西单抗是由人 IgG1-Fc 和鼠 Ig 可变区组成的嵌合体 TNF-α 单抗，阿达木单抗是人源 IgG1 型 TNF-α 单抗，这2 种药物均经静脉注射给药[1]。TNFAI的主要不良反应有注射部位反应、感染、肿瘤、淋巴增殖性疾病、神经脱髓鞘病变以及狼疮样综合征^[1]。由于风湿性疾病多累及生育年龄女性，同时以 TNFAI 为代表的生物制剂应用越来越广泛，TNFAI 对妊娠是否有影响日益受到临床关注。我们复习了2007 年 2 月1日以前 PubMed 中关于上述 3 种 TNFAI对妊娠影响的临床观察文献，现综述如下。......