Anti-inflammatory Activity of 1-docosanoyl Cafferate Isolated from Rhus verniciflua in LPS-stimulated BV2 Microglial Cells

Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as protection of neuronal cells against excitotoxicity, the biological activity of 1-docosanoyl cafferate (DC) has not been examined. The objective of the present study was to evaluate the anti-inflammatory effects of DC, isolated from the stem bark of Rhus verniciflua, on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Pretreatment of cells with DC significantly attenuated LPS-induced NO production, and mRNA and protein expression of iNOS in a concentration-dependent manner. DC also significantly suppressed LPS-induced release of cytokines such as TNF-α and IL-1β . Consistent with the decrease in cytokine release, DC dose-dependently and significantly attenuated LPS-induced mRNA expression of these cytokines. Furthermore, DC significantly suppressed LPS-induced degradation of IKB, which retains NF-kB in the cytoplasm. Therefore, nuclear translocation of NF-kB induced by LPS stimulation was significantly suppressed with DC pretreatment. Taken together, the present study suggests that DC exerts its anti-inflammatory activity through the suppression of NF-kB translocation to the nucleus.     

Evaluation of hydro-alcoholic extract of Eclipta alba for its anticancer potential: an in vitro study

Ethnopharmacological relevance

Eclipta alba is traditionally used as hepatoprotective agent. The study was designed to explore its antiproliferative activity on liver and other related cancer.

Aim of the study

The present study was designed to assess and establish the role of Eclipta alba as anti-cancer agent using HepG2, C6 glioma and A498 cell lines as model system.

Materials and methods

Antiproliferative and cytotoxic effects of the Eclipta alba hydroalcoholic extract (EAE) was determined using MTT assay. The expression level of NF-kB was analysed by western blotting and RT PCR. Gelatin zymography was done for gelatinase matrix metalloproteinases (MMP-2 and 9) analysis.

Results

EAE inhibited the cell proliferation in dose dependent manner in HepG2, A498 and C6 glioma cell lines with an IC50 of 22 ± 2.9, 25 ± 3.6 and 50 ± 8.7 μg/ml, respectively. The expression of MMP (2 and 9) was down-regulated with EAE treatment. DNA damage was observed following 72 h of extract treatment, leading to apoptosis. Additionally, the expression level of NF-kB was evaluated with western blotting and RT-PCR and was found to be down-regulated/inactivated.

Conclusions

The data establish the existence of anti-proliferative, DNA damaging and anti-metastasis properties in EAE which is yet unexplored and hold high therapeutic impact.     

Characterization of a TnMAVS protein from Tetraodon nigroviridis

A growing family of cellular proteins encoding for caspase activation and the recruitment domain (CARD) plays a crucial role in immunity by sensing viral infections and signaling antiviral immune defenses. We obtained a MAVS-like protein (named TnMAVS) from Tetradon nigroviridis, which contains a CARD domain, a pro-rich domain, and a TM domain similar to human MAVS. A fluorescence assay showed that TnMAVS was located in the cytoplasm and near by the membrane, and not the mitochondria in FHM cells. As such, it was considered as a new member of MAVS. The TnMAVS was highly expressed in the liver and muscle of T. nigroviridis. In the spleen, TnMAVS was down-regulated when the fish was treated with polyinosinic:polycytidylic acid or challenged with ISKNV, but was not affected by PGN or LPS. The dual luciferase reporter assay revealed that TnMAVS overexpression resulted in the activation of the interferon-sensitive response element and NF-κB signal pathways. In addition, a characteristic TRAF3-associated peptide PVQD was found in the TnMAVS sequence. Co-immunoprecipitation assays indicated that TnMAVS could interact with zfTRAF3 in eukaryotic cells.     

HSP72过度表达对脂多糖刺激的巨噬细胞中NF-kB活性的影响

目的研究过度表达的热休克蛋白72(heat shock protein,HSP72)对脂多糖(1ipopolysaccharide、LPS)刺激的巨噬细胞中NF-KB活性的影响。方法构建pCD-hsp72重组质粒,转染小鼠巨噬细胞RAW264．7,G418筛选稳定转染hsp72基因的细胞,Westem印迹检测转染细胞HSP72的表达水平及LPS刺激后RAW264．7细胞内HSP72的表达水平、LPS刺激后转染细胞中IKBot的含量和胞核中NF-KB的含量的变化。结果构建pCD-hsp72重组质粒并转入巨噬细胞,获得稳定转染hsp72基因的细胞株,LPS刺激细胞内HSP72表达增加；HSP72可通过减少LPS刺激的巨噬细胞中IkBα的降解而抑制NF-kB活性。结论HSP72能抑制LPS激活的巨噬细胞中NF-kB的活化。     

核因子κB表达与胰腺癌转移及与MMP-2表达的关系

目的： 探讨核因子κB (NF-κB,p65)在胰腺癌（PC）中的表达与淋巴结转移及MMP-2表达的关系。方法：免疫印迹法检测45例PC(淋巴结转移30例，非转移15例)和9例正常胰腺组织(NP)中p65蛋白的表达；RT-PCR检测MMP-2mRNA的表达。 结果：p65，MMP-2在45例 PC组织中的阳性表达率分别为66.7%（30/45）和57.8%（26/45），在30例转移组织中的阳性表达率分别为83.3%（25/30）和73.3%(22/30)，在非转移组织中的阳性表达率分别为33.3%（5/15）和26.7%（4/15）， p65与MMP-2表达呈明显正相关（r=0.743,P<0.05）。结论：NF-κB异常表达与胰腺癌淋巴结转移有关，并与MMP-2正相关，NF-κB 可能通过对下游基因表达的调控，参与了胰腺癌的转移过程。     

YTHDF2 inhibition potentiates radiotherapy antitumor efficacy

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