膜联蛋白B1在猪囊尾蚴免疫逃避作用的研究进展

膜联蛋白是一类有效的内源性调节蛋白,在Ca²⁺存在的条件下与膜磷脂结合,参与细胞活动的多种功能,其与肿瘤发生、自身免疫性疾病、病毒和寄生虫感染等密切相关。作为膜联蛋白家族成员Annexin B1具有独特的氨基酸残基结构,与不同种属的寄生虫入侵特异性宿主密切相关。本文主要阐述了膜联蛋白的种类及膜联蛋白B1在猪囊尾蚴感染宿主机体过程中免疫逃避的作用,旨在探索其猪囊尾蚴感染的免疫逃避机制。深入对膜联蛋白B1和囊尾蚴之间相互关系的进一步了解,以及膜联蛋白B1在寄生虫免疫中更深入的研究,对寄生虫免疫逃避的生物学意义和寄生虫病诊断治疗提供了相关的策略。     

纳米硒对氟化钠诱导小鼠肝组织细胞凋亡的影响

旨在探究纳米硒对氟化钠诱导肝细胞凋亡的影响，40只28日龄昆明小鼠（25 g±2 g）适应性饲养7 d后，随机分为对照组（Control，生理盐水）、氟化钠组（NaF，24 mg·kg^-1）、纳米硒组（Nano-Se，1 mg·kg^-1）、纳米硒治疗组（NaF+Nano-Se，24 mg·kg^-1 NaF+1 mg·kg^-1Nano-Se），每组10只，灌胃给药，试验周期为28 d。通过HE染色评价小鼠肝细胞的病理损伤，Tunel染色评估肝细胞凋亡水平，免疫荧光，Western blot，RT-qPCR检测凋亡相关基因蛋白的表达水平。结果表明：1）纳米硒有效缓解了氟化钠处理引起的肝细胞肿胀、空泡变性以及核碎裂等现象，下调Bax、Caspase3/9及蛋白P53、Caspase3等凋亡发生相关基因的表达，提高了Bcl-2/Bax的表达水平（P<0.05）；2）Tunel染色结果显示，氟化钠处理显著提高了细胞的凋亡率（P<0.01），纳米硒可有效减少细胞凋亡的发生（P<0.05）；3）Cyt-C在氟化钠组表达水平较对照组显著升高（P<0.05），而在纳米硒治疗组的表达呈下降趋势。综上所述，纳米硒通过调控凋亡相关因子的表达能有效缓解氟化钠诱导的小鼠肝细胞凋亡。     

褪黑激素对长毛兔毛囊细胞凋亡的影响

为研究褪黑激素对皖系长毛兔毛囊细胞凋亡的影响及相关机制,选择60只皖系长毛兔,随机均分成4组,1组不埋植褪黑激素(对照),另3组每只兔按25 mg(低剂量组)、40 mg(中剂量组)和55 mg(高剂量组)分别埋植褪黑激素,饲养70 d后,进行组织病理学观察和RNA的转录组高通量测序及q PCR表达分析。结果表明:通过组织切片观察,证实55 mg褪黑激素处理能有效抑制毛囊细胞凋亡,维持细胞正常形态;通过对高剂量组和对照组测序,得到25个差异表达基因,其中有14个基因上调,11个基因下调,涉及通路包括嗜T细胞病毒感染、坏死性凋亡、甲状腺癌、铁死亡、I型糖尿病和胰岛素信号通路,其中坏死性凋亡、铁死亡通路与长毛兔毛囊凋亡相关;选取4个与凋亡相关的差异表达基因进行荧光定量验证,得到新基因MSTRG.3561表达显著上调,SLC25A5表达下调,但差异无统计学意义。     

Identification of a cell-wall peptidase (NlpC/P60) from Nocardia seriolae which induces apoptosis in fathead minnow cells

Nocardia seriolae, a Gram-positive bacterium, is the main pathogen of fish nocardiosis. Protein NlpC/P60 is a cell-wall peptidase and a potential virulence factor of N. seriolae. Subcellular localization research revealed that both NlpC/P60-GFP and NlpC/P60Δsig-GFP fusion proteins were evenly distributed in the whole cell of fathead minnow (FHM) cells. Furthermore, typical apoptotic features, such as nuclear pyrosis and apoptotic bodies, were observed in the transfected FHM cells and grouper spleen cells by the overexpression of protein NlpC/P60. Then, quantitative assays of mitochondrial membrane potential (ΔΨm) value, caspase-3 activity and apoptosis-related gene (Bax, BNIP3, TNF1 and TNF6) mRNA expression were conducted. The results showed that ΔΨm was decreased, caspase-3 was significantly activated, and the mRNA expression of pro-apoptotic genes (Bax and BNIP3) and tumour necrosis factors (TNF1 and TNF6) was up-regulated in NlpC/P60-overexpressed cells. Taken together, the results indicated that the protein NlpC/P60 of N. seriolae might involve in apoptosis regulation. This study may lay the foundation for further study on the function of N. seriolae NlpC/P60 and promote the understanding of the virulence factors and pathogenic mechanism of N. seriolae.     

自噬调节剂对感染日本脑炎病毒小鼠脑部细胞凋亡的影响

旨在研究自噬调控药物对感染日本脑炎病毒（Japanese encephalitis virus，JEV）小鼠脑部细胞凋亡的影响，本试验建立自噬调控药物处理的感染日本脑炎病毒小鼠的动物模型，其中，雷帕霉素为自噬诱导剂，渥漫青霉素及氯喹为自噬抑制剂。实验动物分成8组：DMEM对照组（Control）；JEV感染组（JEV）；JEV+雷帕霉素（Rapamycin）组（JEV+Rapa）；JEV+渥漫青霉素（Wortmannin）组（JEV+Wort）；JEV+氯喹（Chloroquine）组（JEV+CQ）；雷帕霉素组（Rapa）；渥漫青霉素组（Wort）；氯喹组（CQ）。观察不同处理组小鼠的临床症状；透射电镜观察小鼠脑部神经元及胶质细胞的线粒体损伤程度；Tunel染色观察统计小鼠脑部凋亡细胞分布；检测小鼠脑部凋亡因子及凋亡蛋白的表达量。与JEV+Rapa及JEV组相比较，JEV+Wort及JEV+CQ组小鼠出现轻微的神经症状，脑部神经元及胶质细胞线粒体轻度损伤，脑组织较少细胞发生凋亡。不同处理组小鼠脑部凋亡因子及凋亡蛋白的表达量变化差异不显著。综上表明，自噬抑制剂渥漫青霉素和氯喹可以在一定程度上抑制感染日本脑炎病毒小鼠脑组织中细胞凋亡的发生。     

Effects of dissolved oxygen on the growth performance,haematological parameters,antioxidant responses and apoptosis of juvenile GIFT (Oreochromis niloticus)

This study investigated the effects of different dissolved oxygen (DO) levels on the growth performance, antioxidant response and apoptosis of juvenile GIFT (genetically improved farmed tilapia, Oreochromis niloticus). GIFT were fed with five DO levels (1, 2, 3, 4 and 5 mg/L) for 60 days, and the results showed that the final body weight, weight gain rate, specific growth rate and crude protein and crude lipid contents of the fish muscle increased at 5 mg/L DO. The activities of the antioxidant and digestive enzymes were significantly up‐regulated with increasing DO levels. However, the haemoglobin content, number of red blood cells, malondialdehyde content, transaminase activities, glucose content and lactic acid levels decreased at higher DO levels. Furthermore, the cardiomyocyte apoptotic index was significantly decreased with increasing DO levels. Our results show that 5 mg/L DO improved growth performance, promoted antioxidant enzyme activities and reduced liver damage in GIFT.     

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck, one of the most popular edible mushrooms, has attracted great attention due to its delicious taste and healthy properties. However, both its biological effects and the possible mechanism of action have not yet been known. We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck (MPE) against HepG2 human hepatocellular carcinoma cells. Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90% (P<0.01). The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax, Bcl-2 and cleaved caspase-3. On the other hand, MPE might trigger cell cycle arrest at G₀/G₁/S phases by managing p21, Cyclin D1, cyclin-dependent kinases-4 (CDK4) and proliferating cell nuclear antigen (PCNA). Additionally, MPE downregulated TRAF-2 and p-p53, while upregulated p-ASK1 and p-p38. Therefore, it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.     

对INT-FAK信号通路调控脑缺血后神经细胞凋亡的思考

脑缺血性疾病是人类健康的主要杀手之一，相关研究表明，神经细胞的凋亡是造成脑缺血疾病中神经系统损害的主要机制之一，而以整合素-黏着斑激酶（INT-FAK）控制调节的PI3K/PDK/Akt以及Raf/MEK/ERK两条主要信号途径引起的细胞凋亡是其主要作用机制。凋亡过程出现的诸多能加以调控的信号分子，都可以作为治疗脑缺血性损伤的潜在靶点。随着对脑缺血损伤与神经细胞凋亡关联的深入研究，抗凋亡治疗已经成为治疗脑缺血性疾病的重要途径。     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

β-carotene attenuates weaning-induced apoptosis via inhibition of PERK-CHOP and IRE1-JNK/p38 MAPK signalling pathways in piglet jejunum

Weaning may cause oxidative injury, immune response impairment, apoptosis and other injuries in piglets. Oxidative and endoplasmic reticulum stress (ERS) can elicit inflammatory responses, and persistent oxidative and ERS also may lead to apoptotic cascades, which is associated with the pathogenesis of multiple diseases. β-carotene, a natural carotenoid, has potential anti-inflammatory and antioxidant functions. However, the effect of β-carotene on apoptosis in weaned piglets and the detailed molecular mechanism remain unclear. In this study, we found that β-carotene decreased malondialdehyde (MDA) levels and increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in piglet serum. β-carotene could inhibit the mRNA levels of caspase-3 significantly, but had no significant inhibitory effect of the mRNA levels of caspase-9 and caspase-12 in the piglet jejunum. In addition, β-carotene decreased the activation of GRP78, CHOP, and JNK/p38 MAPK and the ratio of Bax/Bcl-2. Furthermore, β-carotene had a significant influence on the activation of ERS and apoptosis-related signals in TG-induced IPEC-J2. In the present study, β-carotene pre-treatment attenuated the ratio of Bax/Bcl-2 and prevented TG-induced increases in the level of PERK-CHOP and IRE1-JNK/p38 MAPK pathway activation in a dose-dependent manner. Overall, these findings indicate that β-carotene may protect weaning-induced apoptosis through inhibiting ERS.