Microglial Cell Activation Increases Saturated and Decreases Monounsaturated Fatty Acid Content,but Both Lipid Species are Proinflammatory

Neuroinflammation is a component of age-related neurodegenerative diseases and cognitive decline. Saturated (SFA) and monounsaturated (MUFA) fatty acids are bioactive molecules that may play different extrinsic and intrinsic roles in neuroinflammation, serving as exogenous ligands for cellular receptors, or endogenous components of cell structural, energetic and signaling pathways. We determined the fatty acyl profile of BV2 microglial cells before and after acute activation with lipopolysaccharide (LPS). We also investigated the effect of SFA and MUFA pretreatment on the production of an invasive, neurotoxic phenotype in BV2 cells. Acute activation of BV2 microglia resulted in an increase in the relative content of SFA (12:0, 16:0, 18:0, 20:0, 22:0, and 24:0 increased significantly), and a relative decrease in the content of MUFA (16:1n7, 18:1n7, 18:1n9, 20:1n9, 24:1n9 decreased significantly). In agreement, the major stearoyl-CoA desaturase (SCD) isoform in BV2 cells, SCD2, was significantly down-regulated by LPS. We next treated cells with SFA (16:0 or 18:0) or MUFA (16:1n7 or 18:1n9), and found that levels of secreted IL6 were increased, as was secreted MMP9-mediated proteolytic activity. To test the functional significance, we treated SH-SY5Y neuronal cells with conditioned medium from BV2 cells pretreated with fatty acids, and found a small but significant induction of cell death. Our findings suggest differential intrinsic roles for SFA and MUFA in activated microglial cells, but similar extrinsic roles for these fatty acid species in inducing activation. Expansion of SFA is important during microglial cell activation, but either supplemental SFA or MUFA may contribute to chronic low-grade neuroinflammation.     

3-<i>epi</i>-bufotalin suppresses the proliferation in colorectal cancer cells through the inhibition of the JAK1/STAT3 signaling pathway

Traditional Chinese medicine (TCM) has been increasingly employed in the last decades in China for both preventing and treating a variety of cancers. 3-epi-bufotalin is an active ingredient of TCM “Chanpi” with anti-tumor potential. However, the effect and mechanism of 3-epi-bufotalin on colorectal cancers were not well disclosed. The present study demonstrated that 3-epi-bufotalin could reduce viability, trigger apoptosis, and block the cell cycle at the G₂/M stage in colorectal cancer cell lines HT29, RKO, and COLO205 in vitro. Moreover, 3-epi-bufotalin inhibited the JAK1/STAT3 signaling pathway. These results indicated the anti-proliferation ability of 3-epi-bufotalin in colorectal cancer cells.     

Hydroxyoctadecadienoic Acids Regulate Apoptosis in Human THP‐1 Cells in a PPARγ‐Dependent Manner

Macrophage apoptosis, a key process in atherogenesis, is regulated by oxidation products, including hydroxyoctadecadienoic acids (HODEs). These stable oxidation products of linoleic acid (LA) are abundant in atherosclerotic plaque and activate PPARγ and GPR132. We investigated the mechanisms through which HODEs regulate apoptosis. The effect of HODEs on THP‐1 monocytes and adherent THP‐1 cells were compared with other C18 fatty acids, LA and α‐linolenic acid (ALA). The number of cells was reduced within 24 hours following treatment with 9‐HODE (p < 0.01, 30 μM) and 13 HODE (p < 0.01, 30 μM), and the equivalent cell viability was also decreased (p < 0.001). Both 9‐HODE and 13‐HODE (but not LA or ALA) markedly increased caspase‐3/7 activity (p < 0.001) in both monocytes and adherent THP‐1 cells, with 9‐HODE the more potent. In addition, 9‐HODE and 13‐HODE both increased Annexin‐V labelling of cells (p < 0.001). There was no effect of LA, ALA, or the PPARγ agonist rosiglitazone (1μM), but the effect of HODEs was replicated with apoptosis‐inducer camptothecin (10μM). Only 9‐HODE increased DNA fragmentation. The pro‐apoptotic effect of HODEs was blocked by the caspase inhibitor DEVD‐CHO. The PPARγ antagonist T0070907 further increased apoptosis, suggestive of the PPARγ‐regulated apoptotic effects induced by 9‐HODE. The use of siRNA for GPR132 showed no evidence that the effect of HODEs was mediated through this receptor. 9‐HODE and 13‐HODE are potent—and specific—regulators of apoptosis in THP‐1 cells. Their action is PPARγ‐dependent and independent of GPR132. Further studies to identify the signalling pathways through which HODEs increase apoptosis in macrophages may reveal novel therapeutic targets for atherosclerosis.     

低剂量率γ射线杀伤肿瘤细胞机制的实验研究

用60Co源以1Gy/min剂量率照射Hela细胞,剂量分别为1、2、5、10、15Gy.用AnnexinV和PI双染法观察凋亡细胞形态;DNA梯形条带证实凋亡存在;克隆形成分析细胞增殖能力.结果显示:(1)Hela细胞凋亡率随照射剂量和时间的增加呈上升趋势,照射后168h组各剂量点凋亡率高于其他各时间组.2Gy以下时凋亡率改变不大,达5Gy时凋亡率显著增加,且达峰值(72.57±2.04)%(P＜0.001).(2)早期凋亡细胞,PS外翻,胞膜呈绿色荧光圈.凋亡晚期出现Annexin V-FITC及PI染色均阳性的外绿内红的细胞图像.坏死细胞则为红色.(3)凋亡细胞碎片呈"梯状"条带.(4)1Gy/min的剂量率照射,剂量为2-15Gy,克隆形成率由(58.95±0.36)%降至(1.67±0.35)%(P＜0.001).表明低剂量率γ射线照射可诱导Hela细胞凋亡,其凋亡率与照射剂量相关,在5Gy时凋亡率最高.     

Polyphenol-rich seaweed (Eucheuma cottonii) extract suppresses breast tumour via hormone modulation and apoptosis induction

The edible red seaweed Eucheuma cottonii is abundantly cultivated for carrageenan production. This study investigated the effects of dietary E. cottonii polyphenol-rich extract (ECME) on breast cancer. In vitro assays showed that ECME was antiproliferative against oestrogen-dependent MCF-7 and oestrogen-independent MB-MDA-231 human breast-cancer cells (IC₅₀ values of 20 and 42 μg/ml, respectively) but was non-toxic to normal cell lines. The ECME (150 and 300 mg/kg BW) was fed to female rats and, after 4 weeks, rat mammary tumour was induced using LA7 cells (inoculated subcutaneously). The ECME inhibited tumour development and erythrocyte lipid peroxidation in the cancer-induced rats, dose-dependently. It showed anti-oestrogenic effects on the rat estrous cycle and serum hormone levels. Electron microscopy and histopathology observations confirmed apoptosis in the rat mammary tumours. The polyphenol-rich ECME was tumour-suppressive via apoptosis induction, downregulating the endogenous oestrogen biosynthesis, and improving antioxidative status in the rats.     

姬松茸多糖对糖尿病大鼠脑组织保护作用及其机制

目的:探讨姬松茸多糖对糖尿病大鼠脑组织保护作用及其作用机制。方法:将Wistar大鼠随机分为5组,A:正常对照组;B:模型组;C:2%姬松茸多糖组;D:4%姬松茸多糖组;E:8%姬松茸多糖组。8周后,HE染色观察脑组织病理变化,琼脂糖凝胶电泳检测细胞凋亡,RT-PCR观察p53基因的表达。结果:与模型组相比,姬松茸多糖治疗组脑组织形态明显改善,脑组织细胞凋亡量明显减少,p53基因的表达明显减少。结论:姬松茸多糖对糖尿病大鼠脑组织有明显保护作用,其机制可能与抑制细胞凋亡及p53基因的表达有关。     

Cancer chemoprevention and chemotherapy: dietary polyphenols and signalling pathways

Prevention of cancer through dietary intervention recently has received an increasing interest, and dietary polyphenols have become not only important potential chemopreventive, but also therapeutic, natural agents. Polyphenols have been reported to interfere at the initiation, promotion and progression of cancer. They might lead to the modulation of proteins in diverse pathways and require the integration of different signals for the final chemopreventive or therapeutic effect. Polyphenols have been demonstrated to act on multiple key elements in signal transduction pathways related to cellular proliferation, differentiation, apoptosis, inflammation, angiogenesis and metastasis; however, these molecular mechanisms of action are not completely characterized and many features remain to be elucidated. The aim of this review is to provide insights into the molecular basis of potential chemopreventive and therapeutic activities of dietary polyphenols with emphasis in their ability to control intracellular signalling cascades considered as relevant targets in a cancer preventive approach.     

Induction of cell apoptosis in 3T3-L1 pre-adipocytes by flavonoids is associated with their antioxidant activity

Obesity is biologically characterized at the cellular level by an increase in the number and size of adipocytes differentiated from fibroblastic pre-adipocytes in adipose tissue. In this study, we focused on the relationship between the influence of flavonoids on cell population growth and their antioxidant activity. The results showed that the inhibition of flavonoids (naringenin, rutin, hesperidin, resveratrol, naringin and quercetin) on 3T3-L1 pre-adipocytes was 28.3, 8.1, 11.1, 33.2, 5.6 and 71.5%, respectively. In oxygen radical absorbance capacity (ORAC) assay, quercetin had the highest ORAC(ROO) value among the six flavonoids tested. Apoptosis assays showed that quercetin increased apoptotic cells in time- and dose-dependent manner. Treatment of cells with quercetin decreased the mitochondrial membrane potential in the courses of time and dose. The cell apoptosis/necrosis assay showed that quercetin increased the number of apoptotic cells, but not necrotic cells. Quercetin treatment of cells caused a significant time- and dose-dependent increase in the caspase-3 activity. Western analysis indicated that treatment of quercetin markedly down-regulated PARP and Bcl-2 proteins, and activated caspase-3, Bax, and Bak proteins. These results indicate that quercetin efficiently inhibits cell population growth and induction of apoptosis in 3T3-L1 pre-adipocytes.     

细胞凋亡的研究

细胞凋亡是一种进化保守的细胞死亡形式,在细胞正常的生理状态中具有极其重要的作用。本文综合概述了细胞凋亡的特点、机理及其检测方法。