中草药复方制剂对青脚麻鸡法氏囊免疫功能、抗氧化性及增殖凋亡基因表达的影响

试验旨在研究由金银花、黄连、蒲公英等药材组成的中草药复方制剂,对青脚麻鸡法氏囊组织结构、抗氧化功能、细胞因子和抗体分泌及增殖凋亡相关基因(PCNA和Caspase-3)表达的影响。采用单因子试验设计,将400羽青脚麻鸡按照体重相近原则分为对照组(基础日粮)和试验Ⅰ-Ⅲ组(分别添加0.5%、1.0%和1.5%中草药复方制剂),每组5个重复(n=20),试验期56 d。结果表明,28日龄时,添加1.0%中草药复方制剂可增加青脚麻鸡法氏囊细胞因子(IL-2和IFN-γ)、IBDV抗体和GSH-Px含量及PCNA mRNA和蛋白表达量(P0.05),降低Caspase-3 mRNA和蛋白表达量(P0.05)。56日龄时,添加1.0%中草药复方制剂可增加青脚麻鸡法氏囊IFN-γ、IBDV抗体和GSH-Px含量(P0.05),降低MDA含量及Caspase-3蛋白表达量(P0.05)。显微观察可见,28和56日龄时,添加1.0%中草药复方制剂可显著增加法氏囊小叶和淋巴小结面积(P0.05),降低法氏囊Caspas-3阳性细胞数量,说明日粮中添加1.0%中草药复方制剂对法氏囊组织结构及免疫抗氧化功能均有明显改善,对法氏囊增殖和凋亡相关基因表达有明显调控作用。文章分别从分子和组织水平揭示中草药复方制剂对青脚麻鸡法氏囊生长发育和免疫功能的影响及可能机制,为新型中草药复方制剂研发及其在青脚麻鸡生产中的应用提供科学依据。     

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。     

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。     

高效氯氰菊酯对小鼠卵巢生殖功能的影响及维生素E的干预作用

为探究高效氯氰菊酯 (beta-cypermethrin, β-CP) 对雌性小鼠卵巢生殖功能的影响及维生素E (vitamin E, VE) 的干预作用，将雌性昆白小鼠随机分为6 组:空白对照组 (花生油处理)、β-CP不同剂量 (10、20、40 mg/kg) 处理组、VE保护组 (20 mg/kg β-CP+20 mg/kg VE) 和VE组 (20 mg/kg VE)，连续灌胃10 d。灌胃结束后取小鼠卵巢组织，观察组织结构的病理变化，采用免疫组化法、蛋白免疫印迹试验及RT-PCR方法检测卵巢中StAR蛋白含量及casp-3、casp-8、INHα和INHβB 基因mRNA表达的变化。结果显示:与对照相比，10、20和40 mg/kg的β-CP处理均使小鼠卵巢组织结构发生了损伤，使组织中StAR蛋白的浓度分别降低了18.8%、36.3%和40.3%，casp-3基因的表达分别升高了16.0%、26.7%和52.9%，INHα基因的表达分别升高了34.5%、83.6%和228.7%，INHβB基因的表达分别升高了7.5%、39.2%和52.7%；20和40 mg/kg的β-CP处理使得casp-8基因的表达分别升高了27.1%和36.7%。上述处理组与对照组的差异均达显著水平 (P＜0.05)。与 20 mg/kg β-CP处理组相比，VE 保护组的StAR蛋白含量也显著增多 (P＜0.05)。研究表明，β-CP对小鼠卵巢具有毒性作用，这与β-CP抑制StAR蛋白的合成，上调casp-3、casp-8、INHα及INHβB基因的表达有关；添加VE对小鼠卵巢有一定的保护作用，这与VE可减弱β-CP对StAR蛋白合成的抑制作用有关。     

加味生脉补心丹对2型糖尿病大鼠心肌损伤的保护作用

目的 探讨加味生脉补心丹（以下简称BXD）对自发性非肥胖2型糖尿病Goto-kakisaki （GK）大鼠心肌损伤的保护作用。方法 8~9周龄雄性Wistar大鼠8只作为空白组，另将40只糖尿病模型GK大鼠按随机数字表将大鼠分为5组，分别为：模型组、西药组（格列齐特缓释片组）、BXD低剂量组、BXD中剂量组、BXD高剂量组。各组大鼠按治疗方法给药10周后处死，腹主动脉采血检测空腹血糖（FPG）、糖化血红蛋白（HbAlc）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA），取左心室行心肌组织HE染色、PAS染色观察病理改变，免疫组化测定心肌凋亡相关蛋白Bax、Bcl-2的表达。结果 与空白组比较，模型组FPG、HbAlc显著升高（P<0.01），SOD、GSH-Px均显著降低（P<0.01），MDA显著升高（P<0.01）；HE染色可见心肌组织水肿坏死、纤维溶解断裂；PAS染色可见大量糖原物质沉积；免疫组化示心肌组织Bax表达增多、Bcl-2表达减少（P<0.01）。与模型组比较，BXD低剂量组FPG降低（P<0.05），西药组和BXD中、高剂量组FPG均显著降低（P<0.01）；西药组HbAlc显著降低（P<0.01），BXD高剂量组HbAlc降低（P<0.05）；西药组与BXD高剂量组SOD、GSH-Px均显著升高（P<0.01）、MDA含量均降低（P<0.05）；西药组、BXD高剂量组Bax表达显著减少（P<0.01），西药组及BXD中剂量组、高剂量组Bcl-2表达显著增多（P<0.01）。结论 BXD对GK大鼠心肌损伤具有保护作用，其机制可能是通过增强抗氧化能力、抑制心肌细胞凋亡，从而起到心肌保护作用。     

加味大柴胡汤对阻塞性黄疸老年大鼠TRPC1表达的影响

目的 探讨加味大柴胡汤在临床上辅助治疗老年阻塞性黄疸的理论机制。方法 通过结扎胆总管的外科手术方法建立阻塞性黄疸老年大鼠模型，实验分为假手术组、模型组、加味大柴胡汤组，予以相应干预，分3、7、10 d共3个时间段，每组10只SPF级老年SD大鼠，在各时间段取老年大鼠肝组织及心脏血，检测AST、ALT及TBA；采用钙离子试剂盒检测老年鼠肝组织匀浆钙离子浓度；采用Western Blot法检测老年鼠肝组织TRPC1蛋白的表达，采用Tunel法观察肝细胞凋亡情况。结果 造模成功后各时间点模型组、加味大柴胡汤组与假手术组比较，ALT、AST、TBA和Ca²⁺浓度均升高，且模型组上升更明显（P<0.01），模型组、加味大柴胡汤组的TRPC1的蛋白含量也均高于假手术组（P<0.01），且加味大柴胡汤组肝细胞的凋亡率低于模型组（P<0.01）。结论 梗阻性黄疸老年大鼠TRPC1表达明显升高，加味大柴胡汤下调TRPC1的表达，可能是其降低钙超载，减少肝细胞凋亡的机制之一。     

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.     

Insights into African swine fever virus immunoevasion strategies

African swine fever(ASF) is an acute and highly contagious disease that causes severe economic losses to the swine industry. ASF is caused by infection of African swine fever virus(ASFV) in domestic pigs, leading to almost 100% mortality. However, no effective vaccines and pharmacologic treatment against ASF are available. ASF poses a severe threat to the swine industry and the economy. Here we summarize potential virus-host cell interaction mechanisms involving the suppression of innate and adaptive immune responses to ASFV entry and infection. These mechanisms include modulation of apoptosis, inhibition of inflammatory responses, reduction of IFN production, inhibition of autophagy, and suppression of MHC-I expression. Insights into immunoevasion strategies by ASFV may shed light on the development of vaccines, as well as preventive and therapeutic drugs.     

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.     

三物白散对Survivin-RNA基因沉默转染胃癌SGC-7901细胞凋亡、迁移的影响

目的 探讨三物白散对Survivin-RNA基因沉默转染胃癌SGC-7901细胞凋亡、迁移的影响。方法 将实验分成6组：空白对照组、LV-RNAi 组、Survivin-RNAi-LV组、空白对照+中药组、LV-RNAi+中药组、Survivin-RNAi-LV+中药组；采用流式细胞仪检测各组细胞的凋亡，划痕实验检测各组细胞的迁移能力。结果 细胞凋亡率：Survivin-RNAi-LV组明显高于空白对照组和LV-RNAi组（P<0.05）；Survivin-RNAi-LV+中药组与空白对照+中药组、LV-RNAi+中药组比较明显较高（P<0.05）；Survivin-RNAi-LV+中药组高于Survivin-RNAi-LV组，差异具有统计学意义（P<0.05）。胃癌细胞的24 h迁移率：Survivin-RNAi-LV组明显低于空白对照组和LV-RNAi组（P<0.05）；Survivin-RNAi-LV+中药组与空白对照+中药组、LV-RNAi+中药组比较，其结果明显低于后两组（P<0.05）；Survivin-RNAi -LV+中药组与Survivin-RNAi-LV组比较，细胞迁移率明显降低，差异有统计学意义（P<0.05）。结论 Survivin基因沉默转染，增强了三物白散诱导胃癌SGC-7901细胞凋亡和抑制其迁移的作用。