宝乐果提取物8,9-糖基化环烯醚萜苷促进人软骨细胞增殖和抑制凋亡的作用

目的观察宝乐果(Borojo)提取物8,9-糖基化环烯醚萜苷(GIG)对体外培养的人骨关节炎软骨细胞促进增殖和抑制细胞凋亡的作用。方法体外培养人骨关节炎软骨细胞,进行软骨细胞鉴定。用不同浓度(25、50、100、200、400μg/ml)的8,9-糖基化环烯醚萜苷培养人骨关节炎软骨细胞,用硫酸酚嗪甲酯(MTS)法、5-乙炔基-2脱氧尿嘧啶核苷(EdU)法、平板克隆形成实验检测细胞的增殖、流式细胞术观察软骨细胞凋亡,用丙二醛(MDA)法、超氧化物歧化酶(SOD)检测、谷胱甘肽过氧化物酶(GSH-Px)检测观察软骨细胞氧化酶的活性,用Western bolt法检测凋亡相关蛋白表达。结果与对照组相比,随GIG浓度增加,细胞活力升高,差异有统计学意义(P<0.01);细胞核分裂增加,差异有统计学意义(P<0.05);细胞克隆数量增多,差异有统计学意义(P<0.05);细胞周期的S期延长,细胞增殖活跃细胞凋亡率减少;MDA含量下降,差异有统计学意义(P<0.05)。SOD含量升高,差异有统计学意义(P<0.05);GSH-Px含量升高,差异有统计学意义(P<0.05)。结论 GIG可能对软骨细胞具有增殖的作用,而且在观察浓度内,细胞毒性小,并可抑制细胞凋亡,提示其有潜在能力可能成为治疗骨关节炎的一种策略。     

CRIP1a switches cannabinoid receptor agonist/antagonist-mediated protection from glutamate excitotoxicity

A shared pathology among many neurological and neurodegenerative disorders is neuronal loss. Cannabinoids have been shown to be neuroprotective in multiple systems. However, both agonists and antagonists of the CB₁ cannabinoid receptor are neuroprotective, but the mechanisms responsible for these actions remain unclear. Recently a CB₁ receptor interacting protein, CRIP1a, was identified and found to alter CB₁ activity. Here we show that in an assay of glutamate neurotoxicity in primary neuronal cortical cultures CRIP1a disrupts agonist-induced neuroprotection and confers antagonist-induced neuroprotection.     

FVB小鼠骨髓源树突状细胞的培养

目的 建立FVB小鼠骨髓源树突状细胞体外培养和扩增方法,观察其形态和特征.方法 在无菌条件下提取FVB鼠骨髓细胞,分离单个核细胞,用IL-4和GM-CSF联合协同诱导下培养,加用磷酸脂多糖刺激.应用光镜下观察树突状细胞的形态,流色细胞仪检测鉴定其生物学特征.结果 联合培养小鼠骨髓细胞3d后,可见细胞形态发生改变,细胞形...     

Spinal cord histopathology of MOG peptide 35-55-induced experimental autoimmune encephalomyelitis is time- and score-dependent

The neuroprotective effects of Jatrorrhizine from Coptidis Rhizoma against hydrogen peroxide (H(2)O(2))-induced rat pheochromocytoma line PC12 injury and its potential mechanisms were evaluated in the present study. When cells were exposed to H(2)O(2) (200 μM) for 12h, there was a significant reduction in cell survival and activity of antioxidant enzyme (SOD and HO-1) and LDH release. In addition, increased ROS production, declined MMP and increased production of malondialdehyde (MDA) were observed. Preincubation of cells with Jatrorrhizine (0.01-10.0 μM) 24h prior to H(2)O(2) exposure markedly elevated cell viability and activities of antioxidant enzyme (SOD and HO-1), prevented LDH release and lipid peroxidation (MDA) production, attenuated the decrease of MMP and scavenged ROS formation. Jatrorrhizine also attenuated caspase-3 activation of the downstream cascade following ROS. Our results suggest that Jatrorrhizine holds potential for neuroprotective effects against H(2)O(2)-induced injury.     

Nucleosome-induced neutrophil activation occurs independently of TLR9 and endosomal acidification: implications for systemic lupus erythematosus

The nucleosome is a major autoantigen known to activate PMN in systemic lupus erythematosus (SLE). TLR9 recognizes bacterial and even mammalian DNA under certain circumstances. Nevertheless, the role of TLR9 in SLE development is still unclear. Since nucleosomes are composed of DNA, we investigated whether TLR9 is required for nucleosome-induced PMN activation. Isolated neutrophils were cultured with nucleosomes, plasma from lupus patients and other stimuli in the presence/absence of various inhibitors. Cells were analyzed by flow cytometry, ELISA and confocal microscopy. We found that nucleosomes circulating in lupus plasma induce the secretion of pro-inflammatory cytokines by PMN. Nucleosomes activate human PMN independently of unmethylated CpG sequences in nucleosomal DNA, leading to IL-8/IL-6/TNF secretion and CD11b up-regulation. Nucleosomes accumulate in the cytoplasm of PMN upon endocytosis, induce TLR9 up-regulation and act synergistically with TLR9 ligands. Nucleosome-induced activation was not inhibited by polymyxin B (PB), chloroquine (CQ), ammonium chloride (AC) or a TLR9 antagonist. Moreover, both PMN isolated from WT and TLR9-KO mice were activated by nucleosomes, as detected by MIP-2 secretion and CD11b up-regulation. Activation occurred therefore independently of endotoxins, endosomal acidification, TLR9 and CpG motifs. TLR9 may thus be differently required in the triggering of nucleosome-induced innate immunity and anti-nucleosome B-cell autoimmunity.     

Acinetobacter baumannii-induced lung cell death: role of inflammation, oxidative stress and cytosolic calcium

A growing body of evidence supports the notion that susceptible Acinetobacter baumannii strain ATCC 19606 induces human epithelial cells death. However, most of the cellular and molecular mechanisms associated with this cell death remain unknown, and also the degree of the cytotoxic effects of a clinical panresistant strain compared with a susceptible strain has never been studied. Due to the role of proinflammatory cytokine release, oxidative stress and cytosolic calcium increase in the cell death-induced by other Gram-negative bacteria, we investigated whether these intracellular targets were involved in the cell death induced by clinical panresistant 113-16 and susceptible ATCC 19606 strains.     

High GMFG expression correlates with poor prognosis and promotes cell migration and invasion in epithelial ovarian cancer

Objective

The aim of this study was to characterize the clinical significance of GMFG, a novel ADF/cofilin superfamily protein, and investigate its role in cell migration and invasion in epithelial ovarian cancer (EOC).

Methods

The expression of GMFG in EOC tissues and ovarian cancer cell lines was evaluated by immunohistochemistry and immunoblotting respectively. The data were statistically analyzed for the associations of GMFG expression with clinicopathologic parameters and survival. In vitro cell migration and invasion assays were performed to determine the role of GMFG in cell migratory behaviors. The effect of GMFG on reorganization of actin cytoskeleton was investigated by immunostaining.

Results

GMFG was overexpressed in EOC. Up-regulated GMFG expression was closely correlated with advanced FIGO stage and chemoresistance of the disease. EOC patients with higher GMFG expression showed poorer progression-free survival (PFS) and overall survival (OS). In vitro cellular assays revealed that GMFG promoted cell migration and invasion. GMFG expression altered actin cytoskeleton organization probably by interacting with the Arp2/3 complex.

Conclusion

GMFG expression independently predicts poorer prognosis in patients with EOC. Ectopic overexpression of GMFG contributes to the malignant biological behavior of ovarian cancer cells.     

The granulocyte colony-stimulating factor (G-CSF) upregulates metalloproteinase-2 and VEGF through PI3K/Akt and Erk1/2 activation in human trophoblast Swan 71 cells

IntroductionAlthough the expression of the granulocyte colony-stimulating factor (G-CSF) and its receptor (G-CSFR) in placental tissues suggests that the cytokine could play a role in placental development, the relevance of G-CSF:G-CSFR interaction in trophoblast cells remains to be studied. Thus, the possible functional role of G-CSF was examined in a human trophoblast cell line (Swan 71 cells).Methods and resultsThe expression of G-CSFR was detected by immunocytochemistry and Western blot assays. G-CSF treatment exerted neither a proliferative nor a protective effect on H₂O₂-mediated cell death in trophoblast cells. Gelatin zymography of supernatants collected from G-CSF-treated cells showed an increment of metalloproteinase-2 (MMP-2) activity. We also found higher MMP-2 and VEGF expression levels in conditioned medium from cells exposed to G-CSF. In addition, it was demonstrated that G-CSF induced the activation of PI3K/Akt and Erk1/2 pathways, which in turn activated NF-kB. By using selective pharmacological inhibitors, it was showed that these pathways are mediating the biological effects produced by G-CSF in Swan 71 cells.Discussion and conclusionWe have demonstrated for the first time that G-CSF increases MMP-2 activity and VEGF secretion in Swan 71 cells through activation of PI3K/Akt and Erk signaling pathways, both contributing to the translocation of NF-kB to the nucleus. These data suggest that G-CSF is involved in the regulation of trophoblast function, and should be considered as a locally produced cytokine probably contributing to embryo implantation and the development of a functional placenta.     

SDF-1对卵巢癌细胞增殖和侵袭能力作用的相关通路研究

目的:探讨PI3K/Akt信号通路抑制剂LY294002、MEK通路抑制剂PD98059对基质衍生因子-1(SDF-1)作用下卵巢癌细胞SKOV3的增殖、侵袭能力的影响,初步探讨SDF-1在卵巢癌中的相关作用通路。方法:细胞免疫化学法检测SDF-1不同作用时间及不同作用浓度对卵巢癌细胞中磷酸化Akt(p-Akt)和ERK(p-ERK)表达的影响。选取对数生长期细胞,分别加SDF-1 100ng/ml、CXCR4中和抗体10μg/ml、CXCR4抑制剂AMD3100 1μg/ml、LY294002 50μmol/L、PD98059 50μmol/L。MTT、Transwell细胞侵袭实验检测细胞的增殖、侵袭能力的变化。结果:随着SDF-1作用时间的延长,p-Akt、p-ERK蛋白表达水平增高;Akt、ERK蛋白的最佳活化时间为30min。作用30min时,随着SDF-1作用浓度的增加,p-ERK1、p-Akt蛋白表达水平增高,差异具有统计学意义(P0.05)。SDF-1可明显促进SKOV3细胞的增殖、侵袭能力(P0.05);但加入CXCR4中和抗体、CXCR4抑制剂AMD3100、通路抑制剂PD98059、LY294002后,SKOV3细胞的增殖、侵袭能力无明显变化(P0.05)。结论:SDF-1对卵巢癌SKOV3细胞的增殖、侵袭能力的增强作用可被PI3K/Akt信号通路抑制剂、MEK通路抑制剂所阻断。SDF-1可能通过Ras/ERK信号转导通路、PI3K/Akt信号通路发挥作用。