Bcl-2和Bax在吗啡依赖雄性大鼠生殖细胞中的表达

目的：探讨B细胞淋巴瘤/白血病-2和人Bcl-2相关x蛋白（Bcl-2、Bax）在吗啡依赖大鼠睾丸生殖细胞中的表达及细胞凋亡可能机制,为治疗阿片类毒品造成的男性性功能减退提供理论依据。方法：以递增法每日给予雄性大鼠皮下注射盐酸吗啡针剂,建立吗啡依赖组。空白对照组注射等量生理盐水。实验成功后将两组大鼠睾丸组织作常规HE染色和免疫组化染色。结果：吗啡依赖组大鼠生精管壁细胞明显地出现上皮层次减少,仅有2~3层,细胞排列疏松,界限模糊,精子细胞和精子数目减少,并发现曲细精管腔内有脱落的生精细胞;免疫组化结果：吗啡依赖组大鼠生殖细胞中bcl-2的阳性表达率明显低于对照组（P〈0.01）,而生殖细胞中bax蛋白的阳性表达率明显高于对照组（P〈0.01）。结论：吗啡依赖可造成雄性大鼠生殖细胞凋亡数量显著增加,其机制可能是通过下调抑凋亡因子Bcl-2,上调促凋亡因子Bax,促进生殖细胞凋亡来实现的。     

鱼藤酮致帕金森大鼠的脑黑质中凋亡相关蛋白Bcl-2、Bax表达改变

目的研究鱼藤酮致帕金森病（PD）大鼠中脑黑质凋亡相关蛋白Bcl-2、Bax表达的改变。方法Wistar大鼠每日颈背部皮下注射鱼藤酮2mg（kg·d）（3～6周）造模,依据所建立的评分体系记录动物行为变化,在行为学有记分并停止给鱼藤酮4、10d时,中脑黑质病理切片免疫组化染色比较黑质区域Bcl-2、Bax的表达。结果在有行为学记分4d时,记4分和8分的大鼠中脑黑质Bcl-2表达均显著减少;所有PD大鼠中脑黑质Bax表达均显著增加;Bcl-2/Bax比率均显著减少;有记分4d时,行为学记分与Bcl-2/Bax比值成负相关性。结论细胞凋亡参与了鱼藤酮帕金森模型大鼠黑质多巴胺神经细胞的损伤。     

双酚A对中国林蛙生精细胞凋亡和Bax、Bcl-2表达的影响

为探讨双酚A(BPA)对两栖动物生精细胞凋亡及相关蛋白Bax和Bcl-2表达的影响.将雄性中国林蛙(Rana chensinensis)分别暴露于10-7、10-6、10-5 mol/L BPA水体中持续3 d、5 d、7 d,取其精巢组织.用原位末端转移酶法(TUNEL)和甲基绿-派诺宁法(Methyl Green-Pyronine)检测生精细胞凋亡,用免疫组织化学方法检测生精细胞的Bax和Bcl-2表达.结果显示,各BPA处理组中国林蛙生精细胞凋亡指数(Apoptotic index,AI)均显著高于对照组,10-6 mol/L与10-7 mol/L BPA处理组生精细胞的AI差异不显著,10-5 mol/L BPA处理组生精细胞的AI与前两组相比显著增高;在同一BPA浓度处理组,生精细胞AI随处理时间的延长而增高.与对照组相比,各BPA处理组Bax表达上调,Bcl-2表达下调,差异均显著;生精细胞AI与Bax/Bcl-2表达呈正相关.这些结果提示,BPA可导致中国林蛙的生精细胞凋亡,而生精细胞凋亡的发生与Bax/Bcl-2表达比值的变化密切相关.     

Phosphorylation of Bcl-2 and Bax Proteins during Hypoxia in Newborn Piglets

Studies indicate that phosphorylated Bcl-2 cannot form a heterodimer with Bax and thus may lose its antiapoptotic potential. The present study tests the hypothesis that graded hypoxia in cerebral tissue induces the phosphorylation of Bcl-2, thus altering the heterodimerization of Bcl-2 with Bax and subsequently leading to apoptosis. Anesthetized, ventilated newborn piglets were assigned to a normoxic and a graded hypoxic group. Cerebral cortical neuronal nuclei were isolated and immunoprecipitated; immune complexes were separated and reacted with Bcl-2 and Bax specific antibodies. The results show an increased level of serine/tyrosine phosphorylated Bcl-2 in nuclear membranes of hypoxic animals. The level of phosphorylated Bcl-2 protein increased linearly with decrease in tissue PCr. The level of phosphorylated Bax in the neuronal nuclear membranes was independent of cerebral tissue PCr. The data shows that during hypoxia, there is increased phosphorylation of Bcl-2, which may prevent its heterodimerization with Bax and lead to increased proapoptotic activity due to excess Bax in the hypoxic brain. Further increased phosphorylation of Bcl-2 may alter the Bcl-2/Bax-dependent antioxidant, lipid peroxidation and pore forming activity, as well as the regulation of intranuclear Ca²⁺ and caspase activation pathways. We speculate that increased phosphorylation of Bcl-2 in neuronal nuclear membranes is a potential mechanism of programmed cell death activation in the hypoxic brain.     

雌激素对大鼠肾上腺皮质中Bcl-2和 Bax蛋白表达的影响

利用免疫组织化学SP法检测摘除卵巢及补充17β-雌二醇后大鼠肾上腺皮质各层Bcl-2和Bax蛋白表达的变化。结果表明,摘除卵巢后肾上腺皮质束状带和网状带Bcl-2和Bax表达都增加;球状带中Bax表达增加,而Bcl-2表达阴性。补充17β-雌二醇后,束状带和网状带中Bcl-2蛋白表达显著降低,Bax在束状带中表达极显著降低,而在网状带中没有阳性产物;在球状带中Bcl-2表达上升,而Bax表达下降。表明雌激素在肾上腺皮质各层发挥着复杂的作用。     

CDC20 regulates the cell proliferation and radiosensitivity of P53 mutant HCC cells through the Bcl-2/Bax pathway

Purpose: The incidence of hepatocellular carcinoma (HCC) is extremely high, and China accounts for approximately 50% of global liver cancer cases. Previous studies reported that CDC20 is involved in the occurrence and progression of a variety of malignant tumors. So, whether CDC20 will affect the development of HCC, we have conducted in-depth research on this.Methods: We selected Hep3B and HepG2 for cell culture, and performed siRNA transfection, lentiviral infection, western blot, MTS determination, cell cycle determination, apoptosis test, immunodeficiency test, clone survival test and subcutaneous parthenogenesis in nude mice.Results: Knockdown of CDC20 greatly enhanced the radiation efficacy on the growth retardation in HepG2, and protein level of CDC20 was decreased for the activation of P53 by radiation. Downregulation of CDC20 combined with radiation can inhibit proliferation, aggravate DNA damage, increase G2/M arrest, and promote apoptosis of HCC cells to a greater extent, and the relative survival fraction of HCC cells was gradually reduced with radiation dose increased in P53 mutated Hep3B cells. After knocking down CDC20 in HCC, Bcl-2 was down-regulated and Bax expression increased. Down-regulation of CDC20 can inhibit further invasion by promoting the radiosensitivity of HCC.Conclusion: In this study, we found that that CDC20 was highly expressed in HCC and participated in radio resistance of HCC cells with P53 mutation Bcl-2/Bax via signaling pathway. This study is the first to present evidence that CDC20 may play a role in improving the efficacy of radiotherapy in HCC.     

妊娠兔胎盘细胞凋亡及凋亡相关基因Bcl-2和Bax表达的动态变化

以新西兰雌兔为动物模型。研究妊娠期间胎盘细胞凋亡及其凋亡调控蛋白Bcl-2和Bax表达的动态变化,基因组DNA凝胶电泳实验检测到妊娠中期和晚期胎盘基因组DNA中出现典型的凋亡特征-DNA梯带,而且DNA断裂值在妊娠早、中、晚期分别为：0.14,0.49和1.43,与妊娠早期相比,妊娠中,晚期胎盘基因组DNA断裂值有显著性增加,TUNEL实验和活化caspase-3的免疫定位实验表明,在妊娠早期胎盘中存在细胞凋亡,而且在各妊娠期中细胞凋亡主要发生于合体滋养层,免疫印迹法分析表明,Bcl-2和Bax随妊娠的进行其表达量明显增加,Bax:Bcl-2比值在妊娠早、中、晚期分别为：0.89,0.91和1.25,呈增加趋势,实验结果说明,在兔正常妊娠中,胎盘合体滋养层细胞发生凋亡,且随妊娠的进行,凋亡细胞数量增多,胎盘细胞凋亡主要与细胞中Bax:Bcl-2的比例相关。     

妊娠兔胎盘细胞凋亡及凋亡相关基因Bcl-2和Bax表达的动态变化

以新西兰雌兔为动物模型,研究妊娠期间胎盘细胞凋亡及其凋亡调控蛋白Bcl-2和Bax表达的动态变化.基因组DNA凝胶电泳实验检测到妊娠中期和晚期胎盘基因组DNA中出现典型的凋亡特征——DNA梯带,而且DNA断裂值在妊娠早、中、晚期分别为：0.14、0.49和1.43,与妊娠早期相比,妊娠中、晚期胎盘基因组DNA断裂值有显著性增加.TUNEL实验和活化caspase-3的免疫定位实验表明,在妊娠早期胎盘中存在细胞凋亡,而且在各妊娠期中细胞凋亡主要发生于合体滋养层.免疫印迹法分析表明,Bcl-2和Bax随妊娠的进行其表达量明显增加,Bax∶Bcl-2比值在妊娠早、中、晚期分别为：0.89,0.91和1.25,呈增加趋势.实验结果说明,在兔正常妊娠中,胎盘合体滋养层细胞发生凋亡,且随妊娠的进行,凋亡细胞数量增多,胎盘细胞凋亡主要与细胞中Bax∶Bcl-2的比例相关.     

镉诱导小鼠睾丸细胞凋亡与Caspase-3、Bcl-2和Bax mRNA的表达

一个月大雄性小鼠24只,随机分为6组,用30 μmol/kg CdCl2作用小鼠睾丸不同的时间(3 h、6 h、12 h、18 h、24 h)后,利用DNA电泳、免疫组化和半定量RT-PCR技术,分析生殖细胞凋亡过程中三种关键物质Caspase-3、Bcl-2、Bax蛋白和mRNA的表达量变化.结果显示:1) DNA各组 (除对照组外)均出现不同程度断裂.2)Caspase-3蛋白表达量一直上升,与对照组相比差异极显著;Bax蛋白在12 h前一直上升,与对照组相比差异显著,12 h后又开始下降,且与对照组相比无显著差异;Bcl-2蛋白在下降,与对照组相比差异显著.3)RT-PCR结果显示Caspase-3基因表达量减少;Bax基因表达量逐渐上升;Bcl-2基因表达量波动很大.综上所述,Caspase-3、Bcl-2和Bax三个基因可能参与了镉应激状态下小鼠睾丸组织细胞的凋亡过程.     

口腔粘膜癌变过程中Bcl-2 及Bax 的表达及临床意义

目的:探讨口腔粘膜癌变过程中口腔粘膜组织中Bcl-2和Bax的表达变化及其临床意义。方法:随机选取我院口腔科就诊的35例口腔粘膜下纤维化(OSF)患者以及15例发生口腔粘膜下纤维化癌变患者为研究对象并分为OSF组和OSF癌变组,并选择15例健康体检者作为对照组。所有研究对象均经统一的口腔科专业医师取口腔内相同部位粘膜组织,通过TUNEL测定其细胞凋亡指数,采用免疫组化法检测其Bcl-2、Bax的表达情况。结果:(1)OSF癌变组、OSF晚期患者的AI均显著高于其他各组(P0.05),OSF癌变组患者AI显著高于OSF晚期组(P0.05),对照组与OSF早期、中期患者AI比较,差异均无统计学意义(P0.05)。(2)OSF癌变组Bcl-2蛋白表达显著高于健康对照组及OSF早、中期患者(P0.05),OSF晚期患者Bcl-2蛋白表达显著高于健康对照组(P0.05);OSF癌变组Bax蛋白表达与健康对照组比较差异有统计学意义(P0.05)。结论:Bcl-2、Bax蛋白的表达均在口腔粘膜癌变过程逐渐上调,可能参与了口腔粘膜癌变的发生和发展。     

Effects of Apigenin on the Expression of LOX-1, Bcl-2, and Bax in Hyperlipidemia Rats

We aimed to investigate the impact of apigenin on LOX-1, Bcl-2, and Bax expression in hyperlipidemia rats and explore the possible molecular pathological mechanism of apigenin in improving hyperlipidemia and preventing atherosclerosis. In hyperlipidemia models, the levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-c) and the LOX-1 protein expression were apparently increased (P<0.01), while the high-density lipoprotein cholesterol (HDL-c) levels and the ratio of Bcl-2/Bax were reduced significantly (P<0.01) in comparison with the standard control group. After the treatment of apigenin, the levels of TC, TG, LDL-c, and the LOX-1 protein expression were noticeably decreased (P<0.01), while the levels of HDL-c and the Bcl-2/Bax ratio were increased (P<0.01). The intima was thickened and had protrusions in the hyperlipidemia model group compared to the normal control group. In comparison with the atherosclerosis model group, the degree of aortic lesions in the low-dose, middle-dose, high-dose groups was alleviated. Apigenin can reduce the level of blood lipid, improve hyperlipidemia, and prevent atherosclerosis in hyperlipidemia rats. The molecular mechanism may be related to inhibiting LOX-1 gene expression and increasing the Bcl-2/Bax ratio.     

Expression of apoptosis-related genes Fas/FasL,Bax/Bcl-2 and Caspase-3 in rat corpus luteum during luteal regression

Using immunohistochemistry, in situ hybridization, Western blot and TUNEL methods, we have studied the expression of Fas/FasL, Bcl-2/Bax and caspase-3 in the corpora lutea (CL) at various stages of pseudopregnant rat induced by injection of PMSF/hCG. The results showed that no apoptotic signal could be observed until Day 14 after hCG injection. Fas weakly expressed in the CL at all the stages increased when luteolysis took place. FasL signal increased dramatically on Day 14 and reached the maximum level on Day 21. The expression of Bcl-2 and Bax was detected in a time-dependent manner. At the early stage of CL development, Bcl-2 expression was stronger, while Bax was low. The expression of Bcl-2 and Bax in the CL was completely reversed. Caspase-3 antigen could be detected throughout all the phases of CL development in a time-dependent fashion, low on Day 2 and reaching the maximum on Day 21. These results suggest that luteal regression at the late phases may be related to cell apoptosis.     

bcl-2核酶与Bax在诱导食管癌细胞凋亡中的协同作用

为了同时调节二种凋亡相关蛋白的表达诱导肿瘤细胞凋亡 ,探索肿瘤基因治疗的可能性 ,同时转入可诱导表达的特异性切割 bcl- 2的核酶基因及 bax基因 ,间接免疫荧光标记法检测 Bcl- 2及Bax蛋白的表达量 ,用 TUNEL、流式细胞术及琼脂糖凝胶电泳检测细胞凋亡 .共转染后 Bcl- 2蛋白表达下降 ,同时 Bax蛋白表达升高 ,导致 30 %左右细胞凋亡 ,并可使细胞对紫杉醇的敏感度增加近4倍 ,使紫杉醇有效作用时间缩短近一倍 .同时调节二个凋亡相关基因可导致细胞凋亡 ,并能有效促进化疗药物诱导的凋亡 .同时校正多个基因的异常表达 ,比仅仅改变单个基因可更有效地达到治疗肿瘤的目的 .     

Expression of apoptosis-related genes Fas/FasL, Bax/Bcl-2 and Caspase-3 in rat corpus luteum during luteal regression

Corpus luteum (CL) is a transient endocrine organ formed from the ovulated follicle. CL produces progesterone and estrogen that are important in preparing the uterine environment for implantation and maintaining gestation. Pregnancy maintains the CL function; otherwise, CL re-gresses rapidly. Cycling formation and regression of CL is essential for initiation of new follicular growth and differentiation, and subsequently ovulation and luteinization[1]. Luteal regression could be divided int…     

Bcl-2/Bax protein expression in heart,slow-twitch and fast-twitch muscles in young rats growing under chronic hypoxia conditions

We have studied the magnitude of apoptosis in heart, slow-twitch skeletal muscle (soleus) and fast-twitch skeletal muscle (gastrocnemius) of rats exposed to 3 weeks in vivo chronic hypoxia. Apoptosis was evaluated biochemically by DNA laddering and by TUNEL and annexin V-staining. The expression of Bax and Bcl-2 proteins was determined by immunohistochemistry and Western blotting.Western blot analysis revealed only a slight difference in Bax expression among the different tissues under normoxic and hypoxic conditions; therefore we can consider that Bax protein is constitutively expressed in muscle tissues. However a singular pattern of Bcl-2 expression was observed among the different tissues under normoxic conditions. Bcl-2 protein was more expressed in fast-twitch glycolytic muscles than in slow-twitch or oxidative muscles with a highest value found in gastrocnemius (4926 ± 280 AU), followed by soleus (2138 ± 200 AU) and a very low expression was displayed in the heart muscle (543 ± 50 AU). After exposure to hypoxia for 21 days (10% O2), Bcl-2 protein expression markedly increased, (44%) in gastrocnemius, (323%) in soleus and (1178%) in heart, with significant differences (p < 0.05 student t-test), reaching a similar threshold of expression in both types of muscles. Furthermore, no sign of apoptosis was detected by TUNEL assay, annexin V-binding assay or DNA electrophoresis analysis. The latter suggested some indiscriminate fragmentations of DNA without apoptosis. In conclusion, we postulate that these protein modifications could represent a adaptative mechanism allowing a better protection against the lack of oxygen in oxidative muscles by preventing apoptosis.     

灯盏花注射液抗新生鼠缺氧缺血性脑损伤的作用及对Bcl-2、Bax蛋白表达的影响

目的：观察灯盏花注射液对新生大鼠缺氧缺血脑损伤（HIBD）的保护作用及对Bcl-2、Bax蛋白表达的影响。方法：采用新生7日龄SD大鼠缺氧缺血性脑损伤模型,设立假手术组、缺氧缺血脑损伤模型组、灯盏花注射液治疗大、中、小剂量组、无菌注射用水对照组。采用硫堇染色、免疫组织化学染色的方法测定各组大鼠海马CA1区神经元密度、组织学分级及凋亡相关基因Bcl-2、Bax蛋白表达情况,并计数各时间点Bcl-2、Bax免疫阳性细胞数目及测定积分光密度值。结果：假手术组,大鼠海马CA1区无锥体细胞缺失,未见明显免疫阳性细胞。与假手术组比较,缺氧缺血脑损伤模型组、无菌注射用水对照组Bcl-2、Bax蛋白表达均于3 d时达到高峰（与其余各时间点比较差别有显著意义P〈0.05）,神经元密度明显降低,组织学分级显著增高,积分光密度值增加。灯盏花治疗组,与无菌注射用水对照组比较,Bcl-2蛋白表达进一步增加,积分光密度值增加;而Bax蛋白表达则减少,积分光密度值降低;神经元密度显著高于对照组,组织学分级明显降低。结论：灯盏花注射液可能是通过上调Bcl-2表达,抑制Bax表达,减轻缺氧缺血引起的神经元凋亡及迟发性神经元死亡。     

白藜芦醇后处理对大鼠脑缺血再灌注损伤海马CA1区Bax、Bcl-2的影响

摘要 目的：探讨白藜芦醇后处理对大鼠脑缺血再灌注损伤Bax、Bcl-2表达的影响。方法：清洁级雄性SD大鼠60只随机分为假手术组（n=12）、I/R组（n=12）、白藜芦醇组（n=36）,白藜芦醇组按不同剂量分为低剂量、中剂量、高剂量组（10 mg/kg、20 mg/kg、40 mg/kg）,每组12只。假手术组：仅暴露大鼠颈外动脉,不做缺血处理;I/R组：采用改良线栓法制备大鼠大脑中动脉缺血再灌注损伤模型（缺血2 h,再灌注24 h）;白藜芦醇组：造模方法同I/R组,在大鼠缺血2h后,将不同剂量白藜芦醇腹腔注射入大鼠体内,比较各组SD大鼠神经功能缺损评分、采用Western blotting法、免疫组化法对大鼠脑组织缺血侧海马CA1区Bax和Bcl-2表达进行比较。结果：白藜芦醇低、中、高剂量组神经功能缺损评分均低于I/R组,随着白藜芦醇剂量的增加,神经功能缺损评分逐渐降低,其中白藜芦醇高剂量组神经功能缺损评分降低最为明显;白藜芦醇组与I/R组相比,不同剂量白藜芦醇组Bax表达逐渐减少,而Bcl-2表达明显增加,其中以白藜芦醇高剂量组改变最为明显。结论：高剂量白藜芦醇可以降低大鼠神经功能缺损评分,减轻脑缺血再灌注损伤,对大鼠脑缺血再灌注损伤具有保护作用,其机制与Bax、Bcl-2的表达有关。     

Altered Bcl-2 and Bax expression is associated with cultured first trimester human cytotrophoblasts apoptosis induced by hypoxia

Preeclampsia is associated with placental hypoxia at early gestation. We therefore investigated the effect of hypoxia on the apoptosis of cultured first trimester human cytotrophoblasts and the expression of apoptosis relevant proteins, Bcl-2 and Bax. First trimester human cytotrophoblasts were isolated and cultured under either standard or hypoxic conditions. Cellular apoptosis was monitored by TUNEL and Annexin V binding, and the expression of Bcl-2 and Bax was determined by Western blot analysis. Apoptosis increased significantly in cytotrophoblasts cultured for 24 h under hypoxic conditions in contrast with those cultured under standard conditions, meanwhile expression of Bcl-2 reduced, and that of Bax increased. These changes suggested that hypoxia induced apoptosis in cultured first trimester cytotrophoblasts with altered Bcl-2 and Bax expression. Further study is needed to explore the role of cytotrophoblasts apoptosis in hypoxia-induced maternal and fetal diseases.     

EPO/EPO-R、Bcl-2、Bax在皮肤血管瘤组织的表达及意义

目的研究促红细胞生成素(erythropoietin,EPO)和促红细胞生成素受体(erythropoietin-receptor,EPO-R)、B细胞淋巴瘤/白血病-2(B-cell lymphoma/Leukemia-2,bcl-2)、促凋亡基因在血管瘤不同时期的表达,探讨其意义及相互关联。方法采用免疫组织化学方法检测人皮肤血管瘤增生期、退化期及正常皮肤组织中EPO/EPO-R、Bcl-2、Bax的表达水平,利用计算机图像分析技术测量不同时期血管瘤组织和正常皮肤组织EPO/EPO-R、Bcl-2、Bax表达的平均光密度和平均阳性面积率。结果 1.EPO/EPO-R在增生期血管瘤内皮细胞的表达明显高于退化期血管瘤内皮细胞和正常皮肤组织血管内皮细胞(P0.01);EPO/EPO-R在退化期血管瘤内皮细胞的表达与正常皮肤组织血管内皮细胞相比,差异无显著性(P0.05)。2.Bcl-2在增生期血管瘤的表达明显高于退化期血管瘤和正常皮肤组织(P0.01);Bcl-2在退化期血管瘤的表达与正常皮肤组织相比,差异无显著性(P0.05)。3.Bax在退化期毛细血管瘤中表达高于增殖期和正常皮肤组织(P0.01),Bax在增殖期中表达高于正常皮肤组织(P0.05)。结论 Bcl-2可能是通过抑制内皮细胞的凋亡,使其增殖;Bax可能通过促进内皮细胞凋亡而抑制血管瘤的增生;EPO/EPO-R可能上调Bcl-2及下调Bax,改变Bcl-2与Bax比值,促进了内皮细胞的增殖。