共查询到19条相似文献，搜索用时 218 毫秒
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旨在研究伪狂犬病病毒(PRV)感染后细胞自噬和凋亡情况,进一步分析鉴定PRV感染条件下与Beclin1互作的猪宿主凋亡家族蛋白。利用免疫印迹(WB)方法检测PRV感染细胞的自噬水平;利用流式细胞术探讨凋亡水平;最后利用免疫共沉淀试验分析PRV感染PK-15细胞中Beclin1与Bcl-2和Bcl-xL的结合水平。结果显示,PRV感染导致LC3-Ⅱ的表达明显增多,而P62显著下降,且AKT(S473)磷酸化水平显著减少;其次,PRV感染诱导细胞凋亡且显著上调剪切型Caspase-9/8的表达水平;最后,免疫共沉淀结果显示PRV感染条件下Beclin1能与Bcl-2和Bcl-xL结合,但结合的水平明显降低。PRV感染诱导的细胞自噬可能依赖于PI3K-Ⅰ/AKT信号通路,PRV感染可能同时激活了线粒体凋亡途经和死亡受体途经从而诱导细胞凋亡,而PRV感染诱导的细胞自噬和细胞凋亡可能通过抑制Beclin1和Bcl-2、Bcl-xL结合而相互关联。 相似文献
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《中国兽医学报》2017,(1):107-111
为研究雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路在镉诱导神经细胞凋亡和相关蛋白Bcl-2、Bax表达中的作用,用醋酸镉和/或雷帕霉素(rapamycin,Rap)染毒PC12细胞24h,流式细胞术检测细胞凋亡率,Hoechst33258荧光染色法检测细胞凋亡形态学变化,Western blot法检测Bcl-2和Bax蛋白表达水平。结果显示:与对照组相比,染毒组细胞凋亡和Bax蛋白表达水平升高,Bcl-2蛋白表达水平降低,Bcl-2/Bax极显著降低(P<0.01);与镉染毒组相比,镉与Rap联合作用组细胞凋亡和Bax蛋白表达水平降低,Bcl-2蛋白表达水平升高,Bcl-2/Bax显著升高(P<0.05)。结果表明:镉可能通过激活mTOR信号通路调节Bcl-2和Bax的表达,从而诱导神经细胞凋亡。 相似文献
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采用Hoechst 33258染色和流式细胞术观察了流感病毒诱导A549细胞凋亡的情况,同时应用Western-blot方法研究了MDM2和p53等蛋白表达的情况.结果表明,100 TCID50/0.1 mL剂量病毒感染A549细胞后,细胞表现出典型的凋亡特征,死亡细胞比例随感染进程逐渐增加.在流感病毒诱导细胞凋亡过程中,MDM2蛋白表达下降,p53和p-p53表达上升,其下游p21蛋白也被激活.可见,p53及其下游p21蛋白参与了流感病毒诱导的A549细胞凋亡,同时通过下调MDM2蛋白表达促进了p53蛋白诱导的细胞凋亡. 相似文献
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为探讨自噬在玉米赤霉烯酮(ZEA)诱导睾丸支持细胞株TM4凋亡中的作用,用不同浓度ZEA(0、5、10、20μmol/L)处理细胞24 h后,流式细胞术检测细胞凋亡率;Western blot检测Bax等凋亡相关蛋白的表达情况;同时检测20μmol/L ZEA联合自噬抑制剂氯喹(CQ)或促进剂雷帕霉素(RAP)后对TM4细胞凋亡率和凋亡相关蛋白及自噬关键蛋白LC-3的变化。结果显示:与对照组相比,5μmol/L ZEA以上染毒组Bax/Bcl-2的比值、cleaved-caspase-3和cleaved-caspase-9蛋白表达量均显著或极显著增加。20μmol/LZEA+c Q可使ZEA诱导的TM4细胞的凋亡率上升,Bax/Bcl-2的比率以及cleaved-caspase-3、cleaved-caspase-9蛋白表达显著下降而自噬标志蛋白LC-3的表达上升;20μmol/LZEA+RAP,可使ZEA诱导的TM4细胞的凋亡率下降;Bax/Bcl-2的比率,cleaved-caspase-3和cleaved-caspase-9蛋白表达及LC-3的表达显著上升。结果表明:在ZEA诱导的TM4细胞发生凋亡的过程中,细胞凋亡率与自噬水平的高低有明显的关系,抑制自噬,ZEA诱导的TM4细胞凋亡率明显升高;促进自噬,ZEA诱导的TM4细胞凋亡率明显降低。研究表明,自噬能够延迟TM4细胞发生凋亡,自噬在ZEA对TM4细胞的毒性损伤中发挥保护作用。 相似文献
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雌激素诱导小鼠胸腺萎缩过程中细胞因子及凋亡相关蛋白基因表达的变化 总被引：1，自引：0，他引：1
本试验应用光镜、电镜、流式细胞术、实时定量PCR技术研究了雌激素诱导小鼠胸腺萎缩过程中胸腺指数、胸腺显微和超微结构、胸腺细胞凋亡率及部分细胞因子和凋亡相关蛋白基因表达的变化。结果注射雌激素后,小鼠胸腺指数显著下降,胸腺中凋亡细胞数量显著上升,细胞因子TGF-β1表达量显著升高,IL-6表达量略为升高,而IL-7表达量显著降低;凋亡相关蛋白Caspase-3、Caspase-9、FADD表达量显著上升,FasL表达量略有上升,而Bcl-2表达量显著降低。表明雌激素可能一方面通过调节胸腺细胞因子抑制胸腺上皮细胞的增殖,另一方面通过激活Caspase级联程序的启动和执行阶段及Fas/FasL凋亡信号通路促进胸腺细胞的凋亡,从而诱导小鼠胸腺萎缩。 相似文献
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《中兽医医药杂志》2019,(3)
目的:探讨钩吻素子(koumine)对H_2O_2诱导的猪肠道上皮细胞(IPEC-J2)损伤的保护作用及其机制。方法:通过MTT法检测koumine对IPEC-J2细胞增殖的影响,并用H_2O_2诱导IPEC-J2细胞建立细胞损伤模型,通过MTT和LDH比色法检测分析koumine保护IPEC-J2细胞免受H_2O_2损伤的效果;生化方法检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;蛋白免疫印迹(western blot)技术检测凋亡相关蛋白bax和bcl-2的表达情况;用流式细胞仪检测氧自由基(ROS)释放量和细胞凋亡率。结果:检测浓度范围内koumine对IPEC-J2细胞没有毒性;koumine可以通过调控IPEC-J2细胞中SOD活性、LDH漏出率、MDA水平和ROS的产生,保护H_2O_2对IPEC-J2细胞的氧化损伤,抑制H_2O_2引起的IPEC-J2细胞凋亡;koumine在抑制H_2O_2引起的细胞凋亡过程中,减少了促凋亡蛋白Bax表达,增加了抑凋亡蛋白Bcl-2表达,使得Bax/Bcl-2的比值在药物作用时随剂量的增加逐渐降低。结论:koumine能够保护IPEC-J2细胞对抗H_2O_2诱导的氧化损伤,具有一定的时间和剂量依赖关系,其作用机制与抑制细胞凋亡有关。 相似文献
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酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白（Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxy genase ActivationProtein,YWHA）家族具有多种生理功能,主要通过调控靶蛋白活性参与调节细胞周期、信号转导、细胞凋亡和细胞代谢等重要生命活动。YWHA家族蛋白高度保守,广泛表达于从植物到动物的各种真核生物中。哺乳动物中存在7种YWHA亚型,分别由不同的基因编码。本文综述了YWHA蛋白结构、在哺乳动物生殖方面以及生殖系统疾病中的功能,为阐明YWHA蛋白在哺乳动物配子发生和生殖过程中的功能提供参考和借鉴。 相似文献
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WANG Ying-qun SHI Bo-mei LI Xun XU Chun-rong LI Mei-zhen TANG Rong-fu WANG Xiao-ye HU Chuan-huo 《中国畜牧兽医》2017,44(7):1967-1974
In this article,firstly,the structure and functions of Bcl-xL protein is briefly introduced. It is believed that Bcl-xL protein as a member of the Bcl-2 family,has the same head folding structures as Bcl-2 family,which plays an important role in suppressing apoptosis of cell and protecting various cells from injury. Then the author introduces the biological properties,functions, mechanisms and clinical application of PTD-FNK protein (a gain-of-function mutant of Bcl-xL). It is thought that PTD-FNK protein is able to rapidly penetrate into cell in vitro and in vivo, and can protect various types of cells from many damage through the inhibition of apoptosis in the path of mitochondrion,so being a potent protein with various therapeutic applications. Finally,the author points out that the difficulty of mass production and practical application of two proteins is that prokaryotic expression of the proteins is inclusion body,which need the complex refolding process in order to obtain soluble proteins,and proteins' loss in the refolding process is great. The priorities and direction of future research are to realize the soluble expression and purify the two proteins in large quantities by optimizing expression conditions of induction,laying the foundation of their application to clinical medicine and cell preservation under room temperature, low temperature and frozen condition. 相似文献
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PTD-FNK蛋白对雄性小鼠生理功能的影响 《畜牧与饲料科学》2022,43(5):13-21
[目的] 探究PTD-FNK蛋白对雄性小鼠生理功能的影响。[方法] 将16只8周龄健康雄性C57BL/6J小鼠随机分为对照组（n=8）和PTD-FNK蛋白组（n=8）,对照组腹腔注射生理盐水,PTD-FNK蛋白组腹腔注射300 μg/（kg·BW）PTD-FNK蛋白,每天腹腔注射给药1次,连续给药7 d;给药期间每天记录小鼠的体重、体温、血糖、采食量、饮水量,计算试验期间总采食量、总饮水量、体增重;于试验第1天和最后1天测量鼻肛距,计算试验前后Lee's指数;试验结束立即处死小鼠,称量心脏、肾脏、肝脏、脾脏和睾丸重量,计算脏器指数;分离小鼠附睾,制备精子悬液,测定精子活力和质膜完整率;利用qPCR法检测睾丸组织中细胞凋亡相关基因Bax和Caspase-3、氧化应激相关基因SOD和GPX1、内分泌相关基因STAR、17β-HSD的mRNA相对表达量。[结果] 与对照组相比,PTD-FNK蛋白组小鼠总采食量极显著（P<0.001）下降,总饮水量显著（P<0.05）下降;Lee's指数、体温、体重、血糖无显著（P>0.05）变化。肝脏指数显著（P<0.05）下降,其他脏器指数无显著（P>0.05）变化。精子活力极显著（P<0.01）升高,精子质膜完整率无显著（P>0.05）变化。睾丸组织中Bax基因mRNA相对表达量极显著（P<0.01）升高,Caspase-3基因mRNA相对表达量无显著（P>0.05）变化,SOD基因mRNA相对表达量极显著（P<0.01）升高,GPX1基因mRNA相对表达量无显著（P>0.05）变化,17β-HSD、STAR基因的mRNA相对表达量无显著（P>0.05）变化。[结论] PTD-FNK蛋白可以抑制雄性小鼠的采食、饮水及肝脏的生长发育,提高小鼠的精子质量。 相似文献
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为研究断喙应激对雏鸡胸腺细胞凋亡及相关凋亡蛋白表达的影响,采用电子显微镜技术、流式细胞术和免疫组织化学技术分析断喙对鸡胸腺淋巴细胞的细胞增殖、分化、凋亡及相关凋亡蛋白(Bcl-2和Bax)表达的影响.结果表明:采用电子显微镜技术观察到断喙对胸腺淋巴细胞产生明显的影响,断喙组胸腺细胞内可看到细胞核明显的应激反应,细胞核凝集,较致密,部分细胞核即将溶解,并出现一定数量的凋亡和坏死细胞;流式细胞仪检测表明对照组和断喙组的胸腺内淋巴细胞都是以G1期为主,但断喙组G1期淋巴细胞数量比试验组高;对照组和断喙组胸腺淋巴细胞凋亡率在断喙后5d时差异显著(P＜0.05);免疫组化结果表明断喙应激没有影响Bcl-2和Bax蛋白在免疫器官内的表达部位;但有降低Bcl 2在胸腺细胞内表达量的趋势;有提高Bax在胸腺细胞内表达量的趋势.结果表明断喙应激促进了雏鸡胸腺淋巴细胞凋亡. 相似文献
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M Colitti 《Anatomia, histologia, embryologia》2012,41(4):237-247
This review focuses on the pro-apoptotic and anti-apoptotic Bcl-2 family members involved in apoptosis, which is the predominant process controlling cell remodelling during post-lactational mammary gland involution. The members of the Bcl-2 protein family, whose expression levels are under the control of lactogenic hormones, internally control this mechanism also during lactation. They can physically interact with each other, sometimes in an antagonistic manner. Mammary glands undergo repeated cycles of structural development, functional differentiation and regression, therefore provide a unique model for investigating this family of proteins that regulate the fate of the secretory cells and consequently milk yield. The involvement of Bcl-2 family members is reviewed in mammary tissue during morphogenesis, at different stages of lactation cycle and in comparison with dairy and laboratory animals. 相似文献
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Madewell BR Gandour-Edwards R Edwards BF Walls JE Griffey SM 《Veterinary pathology》1999,36(6):565-573
The bcl-2 family of genes encodes proteins that influence apoptosis. In the present immunohistochemical study, the topographic distribution of bcl-2 protein was examined in healthy feline fetal, neonatal, and adult tissues, a feline renal cell line, and feline tumors obtained from a veterinary hospital. The topographic distribution of bcl-2 in healthy tissues was similar to that described in human tissues. In lymphoid tissues, follicular mantle cells strongly expressed bcl-2. In complex and differentiating epithelium, bcl-2 expression was detected in stem cell and proliferation zones. Bcl-2 expression was also detected in lower crypts of the intestine and in skin basal layers. The feline Crandell kidney cells expressed bcl-2 diffusely throughout the cytoplasm. Of 180 tumors examined, bcl-2 was expressed almost uniformly in cutaneous basal cell tumors, thyroid adenomas, and mammary carcinomas and in 50% of the lymphomas examined. Bcl-2 may play a role in blocking apoptotic cell death in a broad range of normal feline tissues, whereas dysregulated bcl-2 may extend the life of certain tumors or render certain tumors resistant to therapy because most chemotherapeutic and radiotherapeutic agents eliminate tumor cells by triggering apoptosis. 相似文献
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Expression of Bcl-2 in feline lymphoma cell lines 总被引：1，自引：0，他引：1
Kano R Sato E Okamura T Watanabe S Hasegawa A 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2008,37(1):57-60
BACKGROUND: The Bcl-2 gene is a member of the rapidly expanding Bcl-2 family of genes that regulate apoptosis. Bcl-2 has been shown to repress cell death triggered by a diverse array of stimuli, including chemotherapy and gamma irradiation. OBJECTIVE: The purpose of this study was to determine feline Bcl-2 expression level in feline lymphoma cells using an immunoblot assay with anti-human and anti-canine Bcl-2 monoclonal antibodies. METHODS: About 708 base pairs containing the coding sequence of the feline Bcl-2 gene were transformed into Escherichia coli. The recombinant Bcl-2 was used as a positive control for an immunoblot assay using mouse monoclonal antibodies against human and canine Bcl-2. An immunoblot assay using the monoclonal antibodies was carried out to determine the level of feline Bcl-2 expression in lymphoma and lymphocytic leukemia cell lines. RESULTS: The recombinant feline Bcl-2 protein produced in E. coli had a molecular weight of about 26 kDa and was detected by immunoblot assay by using anti-human Bcl-2 mouse monoclonal antibody. Feline Bcl-2 expression was high in lymphoma cell lines (FL-74-UDC-1 and FT-1) and low in the cell line from peripheral blood mononuclear cells from a healthy cat (FeTJ-1) but not low in freshly isolated peripheral blood mononuclear cells from a healthy cat. The anti-human Bcl-2 mouse monoclonal antibody was found to cross-react with feline Bcl-2. CONCLUSIONS: These results confirm the expression of Bcl-2 in T-cell lymphoma cell lines and indicate that it is suitable to detect feline Bcl-2 using an immunoblot assay. Pending further evaluation, Bcl-2 expression might be useful in the differential diagnosis of feline tumors. 相似文献