坐骨神经损伤对背根节细胞凋亡及细胞凋亡相关蛋白表达的影响

目的：观察大鼠坐骨神经条件性损伤后对背根节细胞凋亡及细胞凋亡相关蛋白表达的影响。方法：将45只成年Wistar大鼠,按手术的不同随机分为3组;正常对照组（A组,n=5）;坐骨神经压榨伤组（B组,n=20);坐骨神经切断组（C组,n=20)。B、C两组又按术后3d,2周,1个月和2个月取材时间分为4个时间组,每组5只大鼠。各时间组取鼠的L5背根神经节后,以TUNEL法检测细胞的凋亡数,以免疫组化技术检测Bcl-2和Bax的表达。结果：正常组及B、C两组伤后3d时未检出凋亡细胞;B、C两组的细胞凋亡率在伤后2周达到高峰,以后随时间的推移而逐渐降低。C组的细胞凋亡率明显高于B组。背根节细胞凋亡相关蛋白Bcl-2及Bax的表达,伤后2周达到高峰。B组中Bcl-2和Bax的表达呈现从低到高而后逐渐恢复正常的规律;而C组的表达始终保持在高水平。结论：细胞凋亡相关蛋白Bcl-2和Bax参与外周神经损伤后背根节细胞凋亡的调节。外周神经的不同损伤方式对背根节细胞凋亡及细胞凋亡相关蛋白的表达影响不同。     

大剂量甲基强的松龙对大鼠急性脊髓损伤后神经细胞Bcl-2表达及细胞凋亡的影响

目的：观察大剂量甲基强的松龙（MP）治疗对大鼠急性脊髓损伤（ASCI）后神经细胞凋亡及凋亡基因Bcl-2的影响。方法：选取48只雌性SD大鼠随机等分为2组,对照组与治疗组,按Nystrom法制备大鼠急性脊髓损伤模型。治疗组伤后30min经腹膜腔注入MP30mg／kg,以后每小时腹膜腔注入MP5．4mg／kg,维持24h;对照组应用生理盐水替代MP,处理方法同治疗组。两组分别于伤后4、8h及1、3、7、14d灌注固定后取材。免疫组织化学检测损伤段脊髓内Bcl-2蛋白表达,TUNEL检测细胞凋亡,染色结果应用图像分析仪进行半定量分析。结果：大鼠ASCI后4h即可见脊髓内TUNEL阳性细胞,8h表达达高峰,此后表达量逐渐下降,14d时仍可见少量阳性细胞。凋亡相关蛋白Bcl-2在伤后4h即可见表达,伤后1d达高峰,伤后14d仍有表达,与对照组相比,治疗组伤后8h、1d和3d时凋亡细胞数减少有统计学意义,伤后8h和1d Bcl-2蛋白表达增高有统计学意义。结论：大剂量甲基强的松龙治疗可抑制大鼠ASCI后神经细胞凋亡,并增加凋亡相关蛋白Bcl-2的表达：     

氯胺酮对慢性坐骨神经压迫性损伤大鼠脊髓背角神经元凋亡及Bax和Bcl-2的表达影响

目的观察氯胺酮（Ket）对慢性坐骨神经压迫性损伤（CCI）大鼠脊髓背角神经元凋亡及凋亡相关蛋白Bax和Bcl-2表达的影响。方法选择雄性SD大鼠40只，随机分为4组，分别为空白对照组，假手术组，CCI组和腹腔注射Ket 50mg／kg治疗组。假手术组，CCI组和Ket治疗组又按术后取材时间的不同分别分为3个亚组，即术后3d组、术后9d组和术后14d组。用TUNEL标记法标记脊髓背角浅层凋亡细胞。应用免疫组织化学方法和免疫印迹（Western-blot）法检测大鼠脊髓背角浅层神经元凋亡相关蛋白Bax和Bcl-2表达情况。结果与空白对照组和假手术组各亚组比较，CCI3d，9d，14d亚组和Ket9d，14d亚组大鼠脊髓背角的凋亡细胞明显增多（P〈0．01）；与CCI3d亚组比较，Ket3d亚组大鼠脊髓背角的凋亡细胞明显减少（P〈0．01）。与空白对照组和假手术各亚组比较，CCI3d，9d，14d亚组大鼠脊髓背角浅层Bax阳性神经元和Bax的表达显著增加（P〈0．01），CCI9d，14d亚组大鼠脊髓背角浅层Bcl-2阳性神经元和Bcl-2的表达明显增加（P〈0．01）。与CCI3d亚组比较，Ket3d亚组脊髓背角浅层Bax阳性神经元和Bax的表达显著减少（P〈0．01），而Bcl-2阳性神经元和Bax的表达显著增加（P〈0．01）。结论Ket能抑制脊髓背角浅层神经元Bax的表达，促进Bcl-2的表达，提示Ket对神经病理性疼痛状态下脊髓背角神经元的凋亡具有保护作用。     

烧伤后大鼠不同骨骼肌凋亡相关蛋白的转录表达变化

目的:通过检测烧伤大鼠不同骨骼肌组织中凋亡相关蛋白的转录变化,探讨细胞凋亡对烧伤后骨骼肌高消耗的影响机制。方法:180只大鼠随机分为5组,通过建立大鼠烧伤模型,分别在烧伤后6h、1天、3天、7天及14天切取大鼠伸趾长肌和比目鱼肌,采用RT-PCR方法测定骨骼肌组织Akt、Bcl-2、Caspase-3的转录表达。结果:伸趾长肌Akt转录表达在伤后前期变化不明显,在第14天显著下降;Bcl-2转录表达在初期轻度上升,在后期下降;Caspase-3转录表达在伤后第7天和第14天显著增高。比目鱼肌Akt转录表达在烧伤后6h、1天、4天、7天无明显变化,在烧伤后14天显著下降;Bcl-2转录表达在烧伤后6h、1天和4天呈逐渐表达显著增强趋势,而后逐渐下降,;Caspase-3转录表达各个时间点无显著变化。结论:在严重烧伤大鼠,快白肌组织凋亡明显,而且随着病程的延长,凋亡越显著;慢红肌组织凋亡不明显,尽管随着病程的延续,有上升的趋势。     

阿魏酸钠抑制缺血性急性肾衰竭肾小管上皮细胞凋亡

目的：探讨阿魏酸钠对缺血性急性肾衰竭大鼠肾脏小管上皮细胞凋亡的影响及机制。方法：将雄性SD大鼠随机分为假手术组、缺血再灌注组、阿魏酸钠冶疗组、SOD冶疗对照组。观察肾组织病理改变；免疫组化、RT—PCR检测Bax，Bcl-2的表达；采用原位DNAg-断末端标记法（TUNEL）检测肾组织细胞凋亡。结果：大鼠肾脏缺血45min再灌注24h，大鼠肾功能损害明显，肾脏组织病理学改变明显，肾小管上皮细胞凋亡明显，肾组织Bax蛋白、mRNA的表达上调，Bcl-2蛋白、mRNA的表达下调。应用阿魏酸钠干预后。改变与上述相反。结论：在缺血性急性肾衰竭中，阿魏酸钠减轻肾脏缺血再灌注桶伤．减轻肾脏细胞凋亡．缓解肾功能桶害．奠作用至少部分通过调控凋亡相关基因Bcl-2家族基因的表达。     

梗阻性黄疸大鼠肾细胞凋亡及bcl-2表达的作用

目的探讨梗阻性黄疸引起肾损伤过程中细胞凋亡的意义。方法32只雄性SD大鼠随机均分为2组，假手术组（SO组）只暴露腹腔但不结扎胆管，胆总管结扎组（BDL组）以双重结扎胆管中间离断方法制成梗阻性黄疸大鼠模型。于术后行光镜下肾脏病理学检查，琼脂糖凝胶电泳检测肾细胞DNA的断裂形式，测定血清中D-Bil、TBA、GOT、GPT、Cr和BUN的含量，检测尿β2-微球蛋白，流式细胞术检测肾脏细胞凋亡率，免疫组化法检测肾组织凋亡抑制基因bcl-2蛋白的表达。结果BDL组大鼠肾脏细胞凋亡率明显高于SO组（P〈0．05），出现细胞凋亡特征性DNA梯状带；其肾脏细胞bcl-2蛋白的表达阳性细胞率明显高于SO组（P〈0．01），并于术后7d达高峰，14d时bcl-2表达阳性细胞率开始下降。结论梗阻性黄疸大鼠肾功能损害过程中存在肾脏细胞凋亡，梗阻性黄疸可反馈性启动肾小管上皮细胞凋亡抑制基因bcl-2的表达。     

肿瘤坏死因子相关凋亡诱导配体及其受体在烧伤大鼠胸腺组织中的表达

目的 了解肿瘤坏死因子（TNF）相关的凋亡诱导配体（TRAIL）及其受体在严重烧伤大鼠胸腺组织细胞异常凋亡中的作用。方法 将50只Wistar大鼠随机分为假伤组（模拟烧伤）lO只和烧伤组40只（设伤后4、12、24、48h 4个时相点）。应用膜联蛋白A5-异硫氰酸荧光素／碘化丙啶双染法，观察大鼠胸腺组织中细胞凋亡的情况；反转录-聚合酶链反应和蛋白质印迹法检测TRAIL的死亡受体5（DR5）、DR4、诱骗受体1（DcR-1）、DcR2在大鼠胸腺组织中的表达。结果 与假伤组大鼠细胞凋亡率[（6．7±0．8）％]比较，烧伤组于伤后4h[（17．1±0．4）％]起增高，12h时[（25．2±1．1）％]达高峰，48h时仍明显高于假伤组（P〈0．05）。烧伤组大鼠胸腺组织中DR5的表达显著高于假伤组，DcR2的表达则显著低于假伤组；其余受体的表达组间相似。结论 严重烧伤后早期大鼠胸腺组织的细胞凋亡明显增加，且胸腺组织中DR5和DcR2的表达异常，提示TRAIL凋亡途径可能参与了病理性细胞凋亡过程。     

IL-6影响骨髓基质细胞凋亡机理的初步探讨

目的通过观察IL-6对体外培养骨髓基质细胞凋亡的影响，探讨骨髓基质细胞凋亡的机理。方法取1月龄SD大鼠的骨髓基质细胞进行体外培养，通过透射电子显微镜观察、bax、bcl-2蛋白免疫组化染色、流式细胞仪检测凋亡细胞周期变化及线粒体跨膜电位改变、RT—PCR法检测凋亡细胞bax、bcl-2 mRNA表达等指标进行观察。结果IL-6组细胞G1期、凋亡率和线粒体膜电位改变均非常显著高于对照组；随着诱导时间的延长，Bcl-2 mRNA表达呈逐渐下降趋势，BaxmRNA表达呈逐渐升高趋势。Bcl-2／Bax：随着诱导时间的延长呈下降趋势。结论IL-6使大量细胞停留在G1期，阻滞细胞进入S期，使DNA合成受阻；IL-6促进凋亡的作用是通过1促进bax从胞浆中移至线粒体膜上而使bax在与bcl-2形成的异二聚体中占据优势来促进线粒体上的PT孔道开放，使内膜离子通道改变，线粒体内膜电位下降或丧失，导致Cyto c等蛋白的释放来调节细胞凋亡。     

FK506对大鼠离体肾细胞凋亡及bcl-2蛋白、bcl-2 mRNA表达的影响

目的观察在含FK506的肾保存液保存的大鼠离体肾脏中bcl-2蛋白、bcl-2mRNA表达及肾细胞凋亡。探讨FK506对离体肾的保护的作用。方法建立离体大鼠肾脏模型，试验组以含FK506的4℃uw保存液保存大鼠离体肾脏，采用免疫组织化学、逆转录-聚合酶链反应（RT-PCR）和细胞凋亡原位末端标记技术，分别检测肾脏保存2、4、8、16h组bcl-2蛋白、bcl-2-mRNA表达。肾细胞凋亡，对照组以不含FK506的4℃ UW保存液保存离体肾脏。结果FK506保存组4、8、16h组bcl-2蛋白、bcl-2mRNA表达明显高于对照组（P〈0．05）；FK506保存组8、16h组肾细胞凋亡指数明显低于对照组（P〈0．05）。结论含FK506肾保存液保存的离体肾脏中bcl-2蛋白、bcl-2mRNA表达增强。肾细胞凋亡指数降低。FK506对肾脏保存过程中的损伤具有保护作用。     

辣椒素对肝星状细胞增殖凋亡的影响

目的 探讨辣椒素(capsaicin)对大鼠肝星状细胞(hepatic stellate cell,HSC) T6细胞系生长的影响及其机制.方法 用CCK-8(Cell Counting Kit-8)法检测终浓度50、100、150、200 μmol/L的辣椒素对大鼠肝星状细胞T6细胞株生长的影响;采用流式细胞仪检测辣椒素对大鼠肝星状细胞T6细胞凋亡率的影响;Western blotting法检测Bcl-2、Bax、细胞色素酶c(Cyt c)蛋白的表达.结果 辣椒素能显著抑制大鼠肝星状细胞T6细胞株增殖,呈时间浓度依赖性(P<0.05).辣椒素作用24 h后T6细胞凋亡率与对照组相比明显升高(P<0.05),与对照组相比,辣椒素作用组Bcl-2蛋白表达量明显下降,bax蛋白的表达明显上升,Cyt c蛋白的表达明显上升,Bcl-2/Bax灰度值明显下降(P<0.05).结论 辣椒素抑制T6细胞增殖并诱导其凋亡,其机制可能是与下调Bcl-2蛋白的表达,上调Bax蛋白的表达,促进Cyt c蛋白的释放有关.     

Adenoviral bcl-2 transfer improves survival and early graft function after ischemia and reperfusion in rat liver transplantation

BACKGROUND: Primary graft dysfunction due to ischemia and reperfusion injury represents a major problem in liver transplantation. The related cell stress may induce apoptosis, which can be suppressed by bcl-2. The purpose of the study was to investigate the effect of adenoviral bcl-2 gene transfer on early graft function and survival in rat liver transplantation. METHODS: An adenoviral construct that transfers bcl-2 under the control of a tetracycline inducible promoter was generated (advTetOn bcl-2) and used with a second adenovirus that transfers the repressor protein (advCMV Rep). Forty-eight hours before explantation, donor rats were treated with advTetOn bcl-2/ advCMV Rep (n=7) and doxycyclin, with the control adenoviral construct advCMV GFP (n=8) or with doxycyclin alone (n=8). Liver transplantation was performed following 16 hours of cold storage (UW). Bcl-2 expression and intrahepatic apoptosis was assessed. Bile flow was monitored 90 min posttransplantation. The endpoint for survival was 7 days. RESULTS: Bcl-2 was expressed in hepatocytes and sinusoidal lining cells. This was associated with a significant reduction of apoptotic sinusoidal lining cells and hepatocytes after 24 hours and 7 days. Bile production was significantly higher following bcl-2 pretreatment. Furthermore, bcl-2 transfer resulted in significantly improved survival (100% vs. 50% both control groups). CONCLUSIONS: Adenoviral bcl-2 transfer results in protein expression in hepatocytes and sinusoidal lining cells resulting in early graft function and survival enhancement after prolonged ischemia and reperfusion injury. The inhibition of apoptosis in the context of liver transplantation might be a reasonable approach in the treatment of graft dysfunction.     

Progesterone differentially regulates pro- and anti-apoptotic gene expression in cerebral cortex following traumatic brain injury in rats

Although the administration of progesterone has been shown to be neuroprotective in experimental models of traumatic brain injury (TBI), the mechanisms for this beneficial effect are still poorly understood. The present study examined the effects of progesterone on mRNA and protein levels of the Bcl-2 apoptosis regulatory genes, bax, bad, bcl-2, and bcl-x(L), in cerebral cortex after TBI. Male Sprague-Dawley rats were subjected to either sham surgery or lateral fluid percussion brain injury of moderate severity (2.4-2.6 atm). Within 1 h post-surgery, progesterone (4 mg/kg) or vehicle (corn oil) administration was initiated for 1-7 days postoperatively. Our results indicate that bax and bad mRNA levels and Bax and Bad protein expression in the ipsilateral, injured cerebral cortex were significantly elevated post-TBI, while mRNA levels of bcl-2 and bcl-x(L) or Bcl-2 and Bcl-x(L) protein expression were not changed. Under the sham-treated condition, progesterone significantly increased mRNA levels of the anti-apoptotic gene, bcl-2, but down-regulated pro-apoptotic gene expression (bax and bad) in cerebral cortex. After TBI, progesterone treatment reduced bax and bad mRNA levels in the ipsilateral cerebral cortex of TBI rats, and decreased Bax and Bad protein levels. In addition, bcl-2 and bcl-x(L) mRNA levels, as well as Bcl-2 and Bcl-x(L) protein expression, were increased by progesterone in TBI injured cortex. These data indicate that one of the neuroprotective mechanisms of progesterone may be related to its differential regulation of apoptotic signals.     

Expression of BCL-2 in liver grafts after adenoviral transfer improves survival following prolonged ischemia and reperfusion in rat liver transplantation

Apoptosis represents a crucial mechanism of ischemia-reperfusion injury after liver transplantation. Bcl-2 may inhibit apoptosis. This study investigates the effect on ischemia/reperfusion injury and survival after rat liver transplantation of adenoviral bcl-2 transfer into donor livers. METHODS: A nonreplicative adenovirus, expressing bcl-2 under control of a tetracyclin-inducible promoter (adv TetOn bcl-2) was used to treat male Lewis rats in combination with a second adenovirus transferring the TetOn repressor protein under control of a cytomegalovirus promoter (advCMVRep). Virus induction was achieved by addition of doxycyclin to the drinking water. Controls were pretreated with a control adenovirus (advCMV GFP) or with doxycycline. Liver transplantations were performed after 16-hour graft storage. Bcl-2 expression was evaluated by Western blot and immunohistology. Survival was monitored for 7 days, and tissue specimens were collected at 24 hours and 7 days post reperfusion. RESULTS: After pretreatment with advTetOn bcl-2/adv CMVRep, intrahepatic bcl-2 expression was evident at 24 hours and 7 days but was absent among controls. Bcl-2 expression was detected in hepatocytes and, to a high degree, in sinusoidal lining cells. TUNEL-positive sinusoidal lining cells were strikingly reduced after bcl-2 transfer (0.1 +/- 0.3 cells/hpf, mean +/- SD) compared to control virus (4.8 +/- 2.3) or doxycyclin-treated grafts (1.3 +/- 0.2); P < .05. After bcl-2 treatment, survival after transplantation was 100%, whereas it was 50% in both control groups (P = .035). CONCLUSION: The study shows the feasibility of transient, doxycyclin-controlled adenoviral gene transfer in a transplantation model. Bcl-2 expression increased survival after ischemia/reperfusion in rat liver transplantation, potentially through protection of sinusoidal lining cells.     

Adenovirus-Mediated bcl-2 Gene Transfer Inhibits Renal Ischemia/Reperfusion Induced Tubular Oxidative Stress and Apoptosis

Ischemia/reperfusion induces oxidative injury to proximal and distal renal tubular cells. We hypothesize that Bcl-2 protein augmentation with adenovirus vector mediated bcl-2 (Adv-bcl-2) gene transfer may improve ischemia/reperfusion induced renal proximal and distal tubular apoptosis through the mitochondrial control of Bax and cytochrome C translocation. Twenty-four hours of Adv-bcl-2 transfection to proximal and distal tubular cells in vitro upregulated Bcl-2/Bax ratio and inhibited hypoxia/reoxygenation induced cytochrome C translocation, O(2) (-) production and tubular apoptosis. Intra-renal arterial Adv-bcl-2 administration with renal venous clamping augmented Bcl-2 protein of rat kidney in vivo in a time-dependent manner. The maximal Bcl-2 protein expression appeared at 7 days after Adv-bcl-2 administration and the primary location of Bcl-2 augmentation was in proximal and distal tubules, but not in glomeruli. With a real-time monitoring O(2) (-) production and apoptosis analysis of rat kidneys, ischemia/reperfusion increased renal O(2) (-) level, potentiated proapoptotic mechanisms, including decrease in Bcl-2/Bax ratio, increases in caspase 3 expression and poly-(ADP-ribose)-polymerase fragments and subsequent proximal and distal tubular apoptosis. However, Adv-bcl-2 administration significantly enhanced Bcl-2/Bax ratio, decreased ischemia/reperfusion induced O(2) (-) amount, inhibited proximal and distal tubular apoptosis and improved renal function. Our results suggest that Adv-bcl-2 gene transfer significantly reduces ischemia/reperfusion induced oxidative injury in the kidney.     

异丙酚对大鼠脑缺血再灌注后脑细胞凋亡和Bcl-2、Caspase-3蛋白表达的影响

脑缺血再灌注后脑损伤的发病机制较复杂,其所致病理损伤包括细胞坏死和凋亡。细胞凋亡与脑缺血再灌注损伤关系密切,是神经元迟发性死亡的主要形式。研究提示与凋亡密切相关的Bcl-2、Caspase-3蛋白等参与了这种迟发性死亡,它们表达的量决定了细胞的生存。本实验采用流式细胞仪、     

Human neuronal apoptosis secondary to traumatic brain injury and the regulative role of apoptosis.related genes

Objective: To observe human neuronal apoptosis secondary to traumatic brain injury, and to elucidate itsregulative mechanism and the change of expression of apoptosis-related genes. Methods: Specimens of brain were collected from cases of traumatic brain injury in humans. The histological and cellular morphology was examined by light and electron microscopy. The extent of DNA injury to cortical neurons was detected by using TUNEL. By in situ hybridisation and immunohistochemistry the mRNA changes and protein expression of Bcl-2, Bax, p53, and caspase 3 p20 subunit were observed. Results: Apoptotic neurons appeared following traumatic brain injury, peaked at 24 hours and lasted for 7 days. In normal brain tissue activated caspase 3 was rare, but a short time after trauma it became activated. The activity peaked at 20-28 hours and remained higher than normal for 5-7 days. There was no expression of Bcl-2 mRNA and Bcl-2 protein in normal brain tissue but 8 hours after injury their expression became evident and then increased, peaked at 2-3 days and remained higher than normal for 5-7 days. The primary expression of Bax-mRNA and Bax protein was high in normal brain tissue. At 20-28 hours they increased and remained high for 2-3 days; on the 7th days they returned to a normal level. In normal brain tissue, p53mRNA and P53 were minimally expressed. Increased expression was detected at the 8th hour, and decreased at 20-28 hours but still remained higher than normal on the 5th day. Conclusions- Following traumatic injury to the human brain, apoptotic neurons appear around the focus of trauma. The mRNA and protein expression of Bcl-2, Bax and p53 and the activity of caspase 3 enzyme are increased.     

Optimal treatment timing to attenuate neuronal apoptosis via Bcl-2 gene transfer in vitro and in vivo

Although Bcl-2 gene transfer can rescue cells from neuronal apoptosis, the temporal relationship between treatment initiation time and effectiveness is unknown. The purpose of present study is to investigate the optimal treatment timing of Bcl-2 gene transfer in saving cells after neural insults. Bcl-2 gene transfer was mediated by recombinant adenovirus carrying human bcl-2 oncogene (Adv-Bcl-2). Adenovirus carrying beta-galactosidase gene (Adv-Bgal) served as a control. A serum withdrawal model of NSC-19 cell culture was used to induce apoptosis in vitro. At various time points before or after serum withdrawal, the motor neuron cells (NSC-19 cells) were infected with either Adv-Bcl-2 or Adv-Bgal. At 72 h after serum withdrawal, the number of apoptotic cells and DNA fragmentation were examined to evaluate the effect of Bcl-2 gene transfer. A weight-drop spinal cord injury model in rats was used as in vivo model. At various time points before or after experimental spinal injury, virus solution, including Adv-Bcl-2 or Adv-Bgal, was injected at the spinal cord in injured rats. The degree of cord injury was measured at 72 h after injury. TUNEL staining was performed to count cells that have undergone DNA damage in sections. Bcl-2 protein overexpression was confirmed by immunostaining both in vitro and in vivo model. In vitro, Adv-Bcl-2 infection produced a less prominent DNA laddering pattern. Adv-Bcl-2 infection between 24 h before and 4 h after serum withdrawal significantly reduced the apoptotic cell death. In vivo Adv-Bcl-2 injection immediately after injury effectively suppressed the injury lesion by blocking DNA fragmentation and irreversible cellular injury. Our data demonstrate that earlier initiation of Bcl-2 gene transfer can produce improved neural cell rescue following neural insults. These results stress important temporal considerations in future gene therapy strategies for spinal cord injury.     

Inhibition of apoptosis reduces immunogeneic potential of adenoviral-treated syngeneic liver grafts

Effects of adenoviral therapy and reduced apoptosis on immune response were investigated in a rat liver transplantation model after prolonged ischemia-reperfusion. Liver donors were treated i.v. either with an adenoviral construct, expressing bcl-2, green-fluorescent-protein, or doxycyclin. Intrahepatic apoptosis was assessed by terminal transferase dUTP nick end labeling assay. The intrahepatic presence of CD4, CD8a, CD163, immunoglobulin (Ig)beta, tumor necrosis factor (TNF)-alpha and myeloperoxidase (MPO) was quantified by realtime polymerase chain reaction at 24 hours and seven days after transplantation. Bcl-2 expression abrogated the TNF-alpha elevation and reduced apoptosis of hepatocytes and sinusoidal endothelial cells as compared to advCMV green fluorescent protein. No effects on CD4, CD8a, CD163 and MPO expression were noticed in bcl-2 pretreated livers, whereas Igbeta was slightly enhanced compared to controls. Adenoviral infected liver grafts trigger an immune response but reduced apoptosis resulted in down-regulation of TNF-alpha. Thus, bcl-2 transfer might simultaneously reduce graft ischemia reperfusion injury and immunogenicity.     

尼卡地平对大鼠Thy1系膜增生性肾小球肾炎的作用及其对肾内Bcl-2/BaX表达的影响

目的：研究尼卡地平诱导系膜细胞调亡在大鼠Thy1系膜增生性肾小球肾炎模型（Thy1模型）中的作用及其与Bcl-2/Bax的表达的关系。方法：（1）应用兔抗大鼠Thy1.1血清建立大鼠Thy1模型;（2）建立模型第7d,腹腔注射尼卡地平（6mg/kg),以等量生理盐水为对照;（3）应用浸条法测定尿蛋白;（4）应用原位末端标记法检测细胞凋亡;（5）应用免疫组化检测Bcl-2/Bax蛋白的表达。结果：注射尼卡地平组尿蛋白程度明显减轻,恢复较生理盐水组快,常规HE染色显示尼卡地平组肾小球内有核细胞数较生理盐水组少,同时肾小球内凋亡细胞数也增多,尼卡地平组肾小球内Bcl-2蛋白阳性细胞数减少,而Bax蛋白阳性细胞数无明显变化,且Bcl-2阳性细胞数与肾小球内细胞数呈负相关。结论：尼卡地平通过诱导系调亡减轻大鼠Thy1模型的肾小球损害,其机制可能与Bcl-2蛋白表达减少有关。