17-β雌二醇对去卵巢大鼠垂体中Bcl-2和Bax表达的影响

为了探讨雌激素对垂体细胞发育和凋亡的影响,利用免疫组化SP法检测摘除卵巢及补充外源性17-β雌二醇后的不同时间大鼠垂体中Bcl-2和Bax表达的变化。结果显示：假去卵巢组（SHAM）Bcl-2和Bax的表达在不同给药时间无显著变化（P〉0.05）;同时间去卵巢（OVX）组与SHAM、去卵巢并注射17-β雌二醇组（OVX＋E2）相比,差异均显著（P〈0.05）;去卵巢给药后6周,OVX组垂体内Bcl-2的表达与之前相比有所增加,而Bax则减少,给药后8周,各组与6周后相比无显著差异（P〉0.05）;同组Bcl-2的表达随时间逐渐减少,4周后最少,Bax的表达则逐渐增加,4周后达到该组的峰值。结果表明,雌激素在垂体的结构维持和功能发挥中起一定的调节作用。     

Inhibitory effect of geinistein on apoptosis in hUVECs induced by MCP-1

AIM: To study the inhibitory effect of genistein on apoptosis in human umbilical vein endothelial cells (hUVECs) induced by monocyte chemotactic protein-1 (MCP-1). METHODS: The hUVECs were cultured in vitro and identified. Growth-arrested hUVECs were stimulated with genistein at different concentrations (0.1 μmol, 1.0 μmol, 10 μmol, 100 μmol) and co-treated with MCP-1 (10 μg/L). The survival rates of hUVECs were detected by MTT assay. The cell cycle and DNA content were detected by flow cytometry. To explore the possible mechanism of the genistein interventions, the expressions of Bcl-2, Fas and Bax proteins were detected by flow cytometry and Western blotting.RESULTS: Genistein increased the survival rate and the level of Bcl-2, inhibited Fas and Bax, decreased the ratios of apoptosis compared with MCP-1-induced hUVECs apoptotic group in a dose-dependent-manner. CONCLUSION: Genistein inhibits the apoptosis induced by MCP-1 and the inhibitory effect was relative to the dose of genistein. Its mechanism might be involved in the down-regulation of Fas and Bax expressions and the up-regulation of Bcl-2.     

Apoptosis and Bcl-2/Bax expression in the early follicles of reproductive women

AIM: To investigate the apoptosis and Bcl-2/Bax expression in the early follicles of women at reproductive age. METHODS: 12 ovarian specimens were collected from reproductive women (aged 23-38 years) undergoing gynaecological operation. Histopathological examination of these specimens was performed to confirm its' morphological normalities. Using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay and immunohistochemistry method, cell apoptosis and Bcl-2/Bax expression were examined in the early follicles including mainly primordial, intermediary and primary follicles. RESULTS: 18.75% of the oocytes were found TUNEL positive in the early follicles, but no granulosa cells in these follicles were found TUNEL positive. Bax expression was detected in 76.07% of the oocytes in the early follicles, but Bcl-2 expression was negative in these oocytes. In addition, Bcl-2/Bax expression were not present in the granulosa cells in early follicles. CONCLUSION: The oocyte apoptosis occurs in the early follicles of reproductive woman, and pro-apoptotic protein Bax may play a role in regulating this process. It suggests that Bax mediated oocyte apoptosis may be the molecular mechanism of the early follicle atresia in the ovaries of reproductive woman.     

银杏叶提取物对大鼠脾脏免疫细胞凋亡相关蛋白Bcl-2和Bax表达的影响

24只SD雌性大鼠随机分为假手术组(A组)、卵巢摘除组(B组)、卵巢摘除并银杏叶提取物(EGb)治疗组(C组),利用免疫组化染色检测脾脏中Bcl-2蛋白和Bax蛋白的表达.结果发现,B组大鼠脾脏内Bcl-2蛋白表达显著减少,而Bax蛋白表达显著增加;C组Bcl-2蛋白表达明显回升(P＜0.01),而Bax蛋白表达明显回落(P＜0.05).表明银杏叶提取物可通过上调Bcl-2蛋白表达,下调Bax蛋白表达,改变Bcl-2与Bax的比值(Bcl-2/Bax)而维护脾脏的免疫自稳状态.     

雌激素对大鼠小脑中Bcl-2和Bax蛋白表达的影响

采用免疫组织化学超敏SP法检测摘除卵巢及用外源性17β-雌二醇治疗后SD大鼠小脑中Bcl-2和Bax蛋白表达的变化.结果显示,摘除双侧卵巢后,SD大鼠小脑皮质及深层核团中Bcl-2的表达有不同程度的降低,而Bax的表达出现不同程度的上升;给予外源性17β-雌二醇治疗后,2种蛋白的比例基本恢复正常.表明,雌激素能够下调小脑皮层及小脑深层核团中Bax蛋白的表达,上调Bcl-2蛋白的表达,对小脑神经元具有保护作用.     

Effect of Vaccinium vitis procyanidin on regulation of glioma cell growth

AIM: To explore the effect of Vaccinium vitis procyanidin on the growth of glioma cells. METHODS: Glioma C6 cells were cultured and divided into control and 10, 20 and 40 μg/L Vaccinium vitis procyanidin groups. The influence of Vaccinium vitis procyanidin on the growth of C6 cells was measured by MTT assay and the observation under inverted microscope. The apoptotic rate was detected by Annexin V/PI staining. The protein expression of Bcl-2 and Bax was determined by immunocytochemistry. The protein levels of Bcl-2, Bax and caspase-3 were also examined by Western blotting. RESULTS: The growth of C6 glioma cells was inhibited by Vaccinium vitis procyanidin at concentrations of 10, 20 and 40 μg/L. The growth was significantly inhibited in 40 μg/L Vaccinium vitis procyanidin group at 24 h and 48 h, and in 20 and 40 μg/L Vaccinium vitis procyanidin groups at 72 h (P<0.01). The density of the cells was decreased when the concentration of Vaccinium vitis procyanidin increased. The apoptotic rate was increased when the concentration of Vaccinium vitis procyanidin increased either. The expression of Bcl-2 was decreased and Bax was increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments. The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased (P<0.05 or P<0.01). The expression of Bcl-2 was decreased (P<0.01), and Bax and caspase-3 were increased after 10, 20 and 40 μg/L Vaccinium vitis procyanidin treatments. The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased (P<0.01). CONCLUSION: Vaccinium vitis procyanidin inhibits the growth of glioma cells by down-regulating Bcl-2 protein and up-regulating Bax protein to activate caspase-3, thus inducing apoptosis.     

Effects of tetrahydroxystilbene-2-O-β-D-glucoside on apoptosis and expression of bcl-2/bax/caspase-3 in HUVECs treated with homocysteine

AIM：To explore the effects of tetrahydroxystilbene-2-O-β-D-glucoside (TSG) from Polygonum multiflorum on the apoptosis and the mRNA expression of bcl-2, bax and caspase-3 in human umbilical vein endothelial cells (HUVECs) treated with homocysteine (Hcy). METHODS：Cultured HUVECs were treated with Hcy (3 mmol/L) to establish a Hcy-damaged model. HUVECs in TSG treated groups were pre-incubated with TSG at concentrations of 1 μmol/L and 10 μmol/L for 2 h before treated with Hcy. Cell nuclear damage was detected by Hoechst 33342 staining. Cell apoptosis was determined by flow cytometry. The mRNA expression of bcl-2, bax and caspase-3 was measured by real-time fluorescence quantitative RT-PCR. RESULTS： After treatment with Hcy at concentration of 3 mmol/L, the nuclear damage and apoptotic rate of HUVECs were higher than that in normal group. The expression of bcl-2 was lower, and the expression of Bax and caspase-3 was higher than that in normal group. On the other hand, pre-incubation with TSG at concentrations of 1 μmol/L and 10 μmol/L decreased the nuclear damage and cell apoptosis, increased the expression of bcl-2, and decreased the expression of bax and caspase-3 as compared with the cells only treated with Hcy. CONCLUSION：TSG reduces the apoptosis of HUVECs induced by Hcy, and the mechanism might be associated with regulating the expression of bcl-2, bax and caspase-3.     

氟对体外培养绵羊成骨细胞Bcl-2和Bax基因表达的影响

目的:探讨氟对体外培养成骨细胞凋亡相关基因Bcl-2和Bax基因表达的影响。方法:采用酶完全消化法分离2～3月龄胎羊颅盖骨成骨细胞,加入不同浓度(0～10-3mol/L)NaF,作用24h后,提取细胞总RNA并反转录成cDNA,通过实时荧光定量PCR方法,检测成骨细胞Bcl-2和Bax基因表达的变化。结果:1.0×10-5mol/LNaF组Bcl-2基因表达水平较对照组高,其他组Bcl-2表达水平均比对照组低;各NaF浓度组Bax基因表达量均比对照组高,且5.0×10-4mol/L组最高;除1.0×10-5mol/LNaF组外,Bcl-2/Bax比值均比对照组低,且5.0×10-4mol/L组最低。结论:高浓度NaF可能增加成骨细胞线粒体膜的通透性,下调Bcl-2 mRNA表达,上调Bax mRNA的表达,可能启动了线粒体凋亡信号转导途径。     

不同能量蛋白水平对绵羊睾丸生精细胞凋亡及相关基因表达的影响

为研究不同能量蛋白对绵羊睾丸生精细胞凋亡及相关基因的影响,选择健康状况良好体重相似的绵羊公羔,将其随机分配到不同能量蛋白水平的3组日粮:100%自由采食组、65%采食组和35%采食组,进行饲喂,当100%自由采食组体重达到35kg后各组取样检测。结果表明,35%采食组中的Bcl-2,Bax mRNA表达量和生精细胞凋亡数量均显著高于其它两组(P<0.05)。65%采食组Bax mRNA表达量和生精细胞凋亡数目均高于100%自由采食组,但是差异均不显著(P>0.05)。100%自由采食组中Bcl-2mRNA表达量高于65%采食组,但是差异不显著(P>0.05)。说明不同能量蛋白水平日粮对绵羊生精细胞凋亡是有影响的,而Bcl-2,Bax参与了此凋亡调控过程。     

黄花补血草多糖的抗肿瘤作用研究

【目的】研究黄花补血草多糖对小鼠肝癌H22细胞生长的抑制作用及对Bcl-2和Bax蛋白表达的影响。【方法】制备小鼠H22实体瘤模型。提取黄花补血草多糖,研究其对小鼠H22实体瘤的抑瘤率,并通过病理切片观察肿瘤细胞的形态学变化。用免疫组化SP法检测黄花补血草多糖对H22肝癌细胞Bcl-2和Bax蛋白阳性细胞数的影响,探讨其抗肿瘤机理。【结果】以阴性对照组为基数,黄花补血草多糖低(120mg/kg)、高(240mg/kg)剂量组及环磷酰胺(CTX,25mg/kg)阳性对照组实体瘤各组抑瘤率分别为31.32%,40.52%和56.61%,与阴性对照组瘤体质量相比显著或极显著减轻。与阴性对照组相比,黄花补血草多糖低、高剂量组Bcl-2蛋白表达分别显著和极显著下调,而Bax蛋白表达显著上调。黄花补血草多糖高剂量组肿瘤细胞表现出坏死如核固缩深染、碎裂或溶解,甚至出现红染区,个别细胞出现染色质浓缩、细胞皱缩、胞膜空泡化和膜周围凋亡小体的形成等凋亡现象。【结论】黄花补血草多糖对小鼠肝癌H22的生长有一定的抑制作用,其机制可能与抑制Bcl-2和提升Bax蛋白表达,从而促进肿瘤细胞凋亡有关。