Expression of apoptosis regulators Bcl-2 and Bax in childhood acute lymphoblastic leukemia

Twenty-five children (19 M:6 F) with newly diagnosed ALL with median age of 5.5 years (1 month-12 years) were enrolled in the study. Apoptosis regulator proteins bcl-2 and bax were measured in all patients using alkaline phosphatase anti-alkaline phosphatase method. Twenty-one patients were positive for bcl-2 and 23 cases for Bax, although expression levels varied. Patients who presented with splenomegaly or hepatomegaly < 5 cm expressed significantly higher levels of bcl-2 and bax protein expression. Neither of age ( < or >10 years), sex, generalized lymphadenopathy, WBC ( < or >50,000/mul) or FAB subtype was associated with high levels of bcl-2 or bax protein expression. Patients with higher mean hemoglobin levels (p = 0.009), high blast % in bone marrow (p = 0.02), immature immunophenotype (p = 0.001) exhibited signifxicantly higher bcl-2 levels. Bcl-2/bax ratio correlated inversely with TLC at presentation (p = 0.022; r = - 0.456) and in B-lineage leukemic cells as compared to T-lineage cells (p = 0.002). Bcl-2/bax ratio did not correlate with any other variable measured. Bcl-2 and bax protein co-express in ALL and high bcl-2/bax ratio correlates with good prognosis features.     

Immunohistochemical expression of bcl-2, bcl-xL, bax, p53 proteins in gastric adenoma and adenocarcinoma]

BACKGROUND/AIMS: The aim of this study was to investigate the immunohistochemical expression of bcl-2, bcl-xL, bax, and p53 proteins according to the pathological parameters such as grade of dysplasia, histological type, depth of invasion, lymph node metastasis, and TNM stage in the gastric adenoma and gastric adenocarcinoma. METHODS: Immunohistochemical staining using monoclonal bcl-2, bcl-xL, bax, p53 antibodies were performed on paraffin embedded specimens from forty-one gastric adenomas and 100 gastric adenocarcinomas. RESULTS: The expression rate of bcl-2 was higher in adenomas (34.2%), especially in high grade dysplasia (52.4%), than adenocarcinomas (2.0%). The expression rate of bcl-xL was higher in adenocarcinomas (55.0%) than adenomas (22%). The expression rate of the bax was higher in adenocarcinomas (58.0%) than adenomas (14.6%). In the adenocarcinoma, the bax expression was significantly related with the depth of invasion, lymph node metastasis, and TNM stage. The expression rate of p53 was higher in adenocarcinomas (64.0%) than adenomas (14.6%). CONCLUSIONS: Bcl-2 protein would be related with the development of gastric adenoma, especially with high grade dysplasia. Bcl-xL and p53 proteins would be involved in the development of relatively early stage of gastric adenocarcinoma but not in tumor progression. Bax protein would be involved in the development of gastric adenocarcinoma and related with depth of invasion, lymph node metastasis, and TNM stage.     

Amount of spontaneous apoptosis detected by Bax/Bcl-2 ratio predicts outcome in acute myeloid leukemia (AML)

The inability to undergo apoptosis is a crucial mechanism of multidrug resistance in acute myeloid leukemia (AML), and the analysis of mitochondrial apoptotic proteins may represent a significant prognostic tool to predict outcome. Bcl-2 and Bax oncoproteins were evaluated in 255 de novo AML patients (pts) by flow cytometry using an anti-bcl-2 monoclonal antibody (MoAb) and an anti-bax MoAb. The results were expressed as an index (bax/bcl-2) obtained by dividing bax mean fluorescence intensity (MFI) and bcl-2 MFI. Lower bax/bcl-2 ratio was associated with French-American-British (FAB) M0-M1 classes (P =.000 01) and CD34 more than 20% (P <.000 01). There were striking inverse correlations between CD34 or CD117 MFI and bax/bcl-2 values (r = -.40, P <.000 001 and r = -.29, P =.000 002), confirming that immaturity is consistent with this index. Moreover, lower bax/bcl-2 levels were correlated with poor-risk cytogenetics (P =.0002). A significant higher complete remission (CR) rate was found in pts with higher bax/bcl-2 levels (79% versus 45%; P =.000 01). Also, both a longer overall survival (OS) and disease-free survival (DFS) were observed in pts with higher bax/bcl-2 levels (P =.000 01 and =.019). Noteworthy, bax/bcl-2 levels accurately predicted the clinical response and outcome of pts with normal or unknown cytogenetics. Indeed, within this subset of 147 pts, higher bax/bcl-2 ratio was significantly associated both with a higher CR rate (86% versus 42%; P <.000 01) and a longer OS (P =.0016). The independent prognostic value of bax/bcl-2 ratio was confirmed in multivariate analysis. Therefore, mitochondrial oncoproteins, such as bcl-2 and bax, represent both sensitive indicators of clinical outcome and potential targets of novel proapoptotic molecules in order to circumvent chemoresistance.     

温阳活血退黄方对阴黄证大鼠肝细胞凋亡及bcl-2和bax蛋白表达的影响

目的：观察温阳活血退黄方对阴黄证大鼠肝细胞凋亡，相关调控基因bcl—2、bax蛋白表达的影响。方法：72只大鼠随机分为6组：正常对照组，阴黄证模型组，阳黄对照组，阴黄证模型加温阳活血退黄方高、低剂量组，菌陈术附汤组。采用TUNEL和免疫组化法检测大鼠肝细胞凋亡和bcl—2、bax蛋白表达。结果：阴黄模型组大鼠肝细胞凋亡程度明显高于阳黄模型组及正常对照组(P＜0．01)，温阳活血退黄方高剂量组肝细胞凋亡程度明显低于阴黄模型组(P＜0．05)。温阳活血退黄方高、低剂量组肝组织bcl—2蛋白表达明显高于阴黄模型组(P＜0．0l，＜0．05)，且其bax蛋白表达明显低于阴黄模型组(P＜0．01，＜0．05)，体现较好量效关系。结论：温阳活血退黄方可能通过促进bcl—2蛋白表达抑制bax蛋白表达阻断阴黄证大鼠肝细胞凋亡，可能是其治疗阴黄证的机制之一。     

Calbindin D28k对帕金森病模型小鼠纹状体凋亡相关蛋白表达的影响

目的观察1-甲基4-苯基1,2,3,6四氢吡啶(MPTP)致帕金森病(PD)模型小鼠黑质多巴胺能神经细胞过表达Calbindin D28k(CB)时,纹状体细胞抗损伤作用机制。方法选择C57BL/6小鼠连续5 d腹腔注射MPTP,构建成功PD模型小鼠30只,随机分为模型组,人类免疫缺陷病毒(HIV)Ⅰ组(注射HIV Ⅰ)和CB-HIV-Ⅰ组(注射CB-HIV-Ⅰ),每组10只,连续6周对各组小鼠行为学检测,Western blot法检测各组小鼠CB,Bcl 2和Bax的表达变化。结果与模型组和HIV-Ⅰ组比较,CB-HIV-Ⅰ组小鼠各时间点移动格子次数,第1、2、5和6周站立次数,第6周时游泳和悬挂时间,差异有统计学意义(P0.05,P0.01);CB-HIV Ⅰ组小鼠中脑黑质中CB的表达量显著升高(P0.05),纹状体细胞中Bcl-2的表达量亦明显升高(P0.01),而Bax的表达量明显降低(P0.01)。模型组和HIV-Ⅰ组上述指标差异无统计学意义(P0.05)。结论黑质多巴胺能神经细胞过表达CB时,纹状体细胞Bcl-2/Bax表达上调,提示其与纹状体细胞抗凋亡能力增强有关。     

Markers of cell proliferation, apoptosis, and angiogenesis in thyroid adenomas: a comparative immunohistochemical and genetic investigation of functioning and nonfunctioning nodules.

OBJECTIVE: To perform (i) an immunohistochemical investigation of cell proliferation, apoptosis, angiogenesis, and malignancy markers in 15 functioning and 15 nonfunctioning thyroid adenomas, and in normal adjacent tissue, and (ii) a genetic analysis of thyroid-stimulating hormone receptor (TSH-r), Gsalpha, and RAS mutations in the same group of adenomas, in order to describe their expression within tissues and to correlate them with the hormonal functioning. DESIGN: Thirty patients who underwent surgery for a solitary thyroid nodule were included in the study. Adenomas and normal adjacent tissues were evaluated by immunohistochemistry using the following antibodies: MIB-1 for proliferative activity, bcl-2 and mutant p53 for apoptosis control, vascular endothelial growth factor-A (VEGF-A) for angiogenic activity, and galectin-3 as a marker for malignancy. To calculate microvascular density, "hot spots" were selected and defined by cells positive for CD34 staining. Genetic analysis for TSH-r, Gsalpha, and H-, K-, and N-RAS mutations was performed on adenoma specimens. MAIN OUTCOMES: Our results evidenced that a proportion of both functioning and nonfunctioning adenomas showed immunohistochemical phenotypes similar to normal adjacent tissue. No differences were found between functioning and nonfunctioning thyroid adenomas with regard to the expression of markers associated to angiogenesis (VEGF-A, microvascular density) and apoptosis control (mutant p53, bcl-2). All adenomas resulted negative for galectin-3 immunostaining. MIB-1 was the only marker showing a substantial difference of expression between the two groups of adenomas. TSH-r mutations were found in 12 out of 15 functioning adenomas, whereas the absence of Gsalpha and H-, K-, and N-RAS mutations was demonstrated in all adenomas. CONCLUSIONS: Our data suggest that the differences between functioning and nonfunctioning thyroid adenomas are restricted to the genetic mutations of the TSH-r, to the hormonal status of tumors, and to the proliferative activity, not involving markers of apoptosis control and angiogenesis.     

Loss of Bcl-2 expression in Barrett''s dysplasia and adenocarcinoma is associated with tumor progression and worse survival but not with response to neoadjuvant chemoradiation

Esophageal adenocarcinoma arising on a background of Barrett's esophagus is increasing in incidence. A molecular understanding of both the progression of Barrett's esophagus and the factors determining the response of adenocarcinoma to neoadjuvant therapy is required, and this study focused on the role of proteins regulated by the bcl-2 family of genes, which are important regulators of programmed cell death (apoptosis). In total, 48 patients (36 men, 12 women) with Barrett's adenocarcinoma were studied. All patients received preoperative chemoradiotherapy followed by surgery. Bcl-2, bax and bcl-x protein expression were detected by standard avidin-biotin peroxidase method. Bcl-2, bax and bcl-x expression were detected in 84%, 80%, and 76%, respectively, of normal squamous mucosa. An increasing degree of dysplasia in Barrett's mucosa both before and after chemoradiotherapy was significantly associated with a reduction of bcl-2 expression (P = 0.03 and 0.009, respectively). Bcl-2 expression was significantly associated with tumor differentiation (P = 0.03) and a trend towards earlier T stage (P = 0.08), but not with nodal status. Pre-therapeutic bcl-2, bax and bcl-x protein expression (27%, 75%, and 87.5%, respectively) were not associated with tumor response or resistance to therapy. Bcl-2-positive patients had a significantly improved survival compared with bcl-2-negative tumors. A significant reduction of bcl-2 expression is associated with the progression of Barrett's mucosa to adenocarcinoma. Bcl-2 expression was associated with improved survival. Preoperative chemoradiotherapy induces expression of bax and bcl-x protein. The pretreatment expression of bcl-2 and related proteins did not predict response or resistance to neoadjuvant chemoradiotherapy, suggesting that regulators of apoptosis alone do not determine the response of Barrett's adenocarcinoma to neoadjuvant therapy.     

螺内酯和缬沙坦对高血压大鼠心肌细胞凋亡相关蛋白的影响

目的探讨螺内酯和缬沙坦对肾血管性高血压大鼠左心室细胞凋亡调控相关蛋白P53、Bax及Bcl-2表达的影响。方法选取SD大鼠46只制成肾血管性高血压大鼠模型后,随机分为高血压组(N组,12只)、缬沙坦组(V组,11只)、螺内酯组(S组,12只)、螺内酯加缬沙坦组(S+V组,11只),另选10只未造模大鼠为假手术组(C组),各组治疗10周后行超声心动图检查,测定大鼠颈动脉压,应用免疫组织化学法检测P53、Bax及Bcl-2蛋白的表达。结果治疗10周后,与C组比较,N组大鼠收缩压明显升高(P0.01);与N组比较,V组、S+V组大鼠收缩压明显降低(P0.05);与C组比较,N组大鼠Bax、Bcl-2和Bax/Bcl-2明显升高,S组大鼠Bax和Bax/Bcl-2明显升高(P0.05,P0.01);与N组比较,S组大鼠Bax、Bax/Bcl-2和V组、S+V组大鼠Bax、Bcl-2、Bax/Bcl-2均明显下降(P0.05,P0.01);与S组比较,V组、S+V组大鼠Bax、Bax/Bcl-2明显下降(P0.05,P0.01)。结论两肾一夹肾血管性高血压大鼠左心室肥厚形成过程中,Bax过表达不完全依赖P53调节。缬沙坦可以通过与血管紧张素Ⅱ1型受体结合,抑制心肌细胞凋亡相关蛋白P53、Bax及Bcl-2的表达及左心室肥厚的发生。螺内酯可以通过与醛固酮受体结合明显降低P53及Bax的表达,降低Bax/Bcl-2比值,部分地逆转左心室肥厚。     

促红细胞生成素预处理在心肌缺氧复氧损伤中对凋亡相关基因表达影响的研究

目的:观察促红细胞生成素(erythropoietin,EPO)预处理在大鼠心肌缺氧复氧损伤(hypoxia/reoxygenation,H/R)中的抗凋亡效应以及对凋亡相关基因bcl-2及bax表达的影响。方法:Wistar成年雄性大鼠60只,分为2组,分别为对照组,EPO预处理组(EPO组),每组30只,EPO组大鼠经腹腔注射5000U/kg重组人促红细胞生成素(Recombinant human erythropoietin,RHuEPO),对照组注射同体积生理盐水。2组各15只大鼠于给药24h后,检测各项指标,其余15只大鼠置于常压缺氧环境中(O27%)12h后,移至常压常氧环境中2h,予H/R损伤,之后采取心肌标本,DNA末端转移酶标记法(TUNEL法)检测心肌细胞凋亡,免疫组化检测凋亡效应酶胱天蛋白酶-3(caspase-3)、凋亡相关基因B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia-2,bcl-2)及B细胞淋巴瘤/白血病相关x蛋白(Bcl-associated x protein,bax)蛋白表达水平,计算bcl-2/bax比值。结果:H/R损伤前2组心肌细胞凋亡率,caspase-3以及bax蛋白表达水平无显著性差异(P>0.05),EPO组bcl-2蛋白表达水平、bcl-2/bax比值显著高于对照组(P<0.01)。H/R损伤后2组心肌细胞凋亡率,caspase-3、bax及bcl-2蛋白表达水平显著高于损伤前(P<0.01),而bcl-2/bax比值显著低于损伤前(P<0.01),EPO组的心肌细胞凋亡率、caspase-3蛋白表达水平显著低于对照组(P<0.05,P<0.01),而bcl-2蛋白表达水平以及bcl-2/bax比值显著高于对照组(P<0.05,P<0.01),2组bax蛋白表达水平差异无显著性(P>0.05)。结论:EPO预处理在心肌H/R损伤中具有显著的抗凋亡效应;这一效应与其上调抗凋亡基因bcl-2的表达有关。     

Detection of bcl-2 and bax expression and bcl-2／JH fusion gene in intrahepatic cholangiocarcinoma

AIM: To investigate the relationship between bcl-2 gene and its related protein bax and intrahepatic cholangiocellular carcinoma (CCC). METHODS: Semi-nested in situ PCR (SNISPCR) and immunohistochemistry were performed to detect bcl-2/JH fusion gene and bcl-2, bax protein expression in 29 cases of CCC. RESULTS: No bcl-2/JH fusion gene was found in all cases of CCC, 72.4% of 29 cases expressed bcl-2 protein. Bcl-2 protein expression was related to histopathological grades (P<0.05). There was no corresponding relationship between bcl-2/JH fusion gene formation and bcl-2 protein expression in CCC (P<0.05). Bax was expressed in 10.3% of 29 cases. The ratio of bcl-2 to bax in normal liver tissues (3.5 to 1) was different from that in tumor tissues (7.0 to 1). CONCLUSION: It is suggested that bcl-2/JH fusion gene formation is not a frequent event and may not play an important role in the pathogenesis of CCC. However, aberrant ratio of bcl-2 to bax protein expression may be involved in the course of tumorigenesis of CCC. Abnormal bcl-2 protein expression may not be solely resulted from bcl-2/JH fusion gene.     

重度妊娠高血压综合征胎盘组织中细胞凋亡的临床观察与研究

目的研究细胞凋亡及其凋亡基因Bax、bcl-2和FAS在重度妊娠高血压综合征(妊高征)患者发生、发展中的作用和意义。方法选择2010年4月—2011年4月我院40例分别为正常晚孕妇女(对照组20例)和重度妊高征患者(观察组20例)的胎盘绒毛膜组织进行分析,采用流式细胞计量术检测方法检测细胞凋亡,促进和抑制凋亡基因(bax、bcl-2)及FAS表达意义。结果对照组胎盘绒毛中胎盘细胞滋养细胞、合体滋养细胞中凋亡指数分别为(1.2±0.9)%、(41.9±1.8)%;bax阳性率分别为(1.1±0.8)%、(29.1±9.8)%;bcl-2阳性率分别为(2.1±0.9)%、(22.9±0.9)%;FAS的表达指数是(5.61±2.79)%。观察组胎盘细胞滋养细胞、合体滋养细胞的凋亡指数分别为(4.4±1.3)%、(44.3±1.5)%;bax阳性率分别是(2.3±0.8)%、(41.5±11.8)%;bcl-2阳性率分别是(3.1±0.9)%、(23.4±7.9)%;FAS的表达指数是(4.01±1.83)%。观察组中细胞凋亡明显增高(P<0.01),bax表达也明显增强(P<0.01)。结论细胞凋亡及其调节基因的表达间具有一致性;在妊高征中起重要的作用;细胞凋亡、bax、bcl-2表达间的变化影响妊娠妇女与重度妊高征患者胎盘的结构与功能,同时我们通过对细胞凋亡的调节而对妊高征的演进过程产生影响。     

Expression of Bcl-2 and Bax in the human corpus luteum during the menstrual cycle and in early pregnancy: regulation by human chorionic gonadotropin

To investigate the relationship between apoptosis and the Bcl-2/ Bax system in the human corpus luteum (CL), the frequency of apoptosis and expression of Bcl-2 and Bax were examined in the CL during the menstrual cycle and in early pregnancy. In situ analysis of DNA fragmentation showed that the number of apoptotic cells was much greater in the regressing CL than that in the midluteal phase CL, whereas there were almost no apoptotic cells in the CL of early pregnancy. Immunohistochemistry revealed that Bcl-2 expression was observed in the luteal cells in the midluteal phase and early pregnancy, but not in the regressing CL. In contrast, Bax immunostaining was observed in the regressing CL, but not in the midluteal phase and early pregnancy. bcl-2 messenger ribonucleic acid (mRNA) levels in the CL during the menstrual cycle were highest in the midluteal phase and lowest in the regressing CL. In the CL of early pregnancy, bcl-2 mRNA levels were significantly higher than those in the midluteal phase. In contrast, bax mRNA levels were highest in the regressing CL and remarkably low in the CL of early pregnancy. Western blot analyses revealed that Bcl-2 expression was significantly lower in the regressing CL than in the midluteal phase and early pregnancy, and that Bax expression was, in contrast, significantly higher in the regressing CL than in the midluteal phase and was remarkably low in the CL of early pregnancy. When corpora lutea of the midluteal phase were incubated with hCG, hCG significantly increased the mRNA and protein levels of Bcl-2 and significantly decreased those of Bax. In conclusion, Bcl-2 and Bax may play important roles in the regulation of the life span of the human CL by controlling the rate of apoptosis. hCG may act to prolong the life span of the CL by increasing Bcl-2 expression and decreasing Bax expression when pregnancy occurs.     

自身免疫性甲状腺疾病甲状腺组织中bcl-2家族蛋白的表达

目的研究凋亡相关基因bcl鄄2家族蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在自身免疫性甲状腺疾病(AITD)甲状腺组织中的表达特征及与AITD发病机制之间的内在联系。方法以甲状腺腺瘤旁正常甲状腺组织为对照(C组,20例),采用免疫组织化学ElivisionTM二步染色法检测凋亡相关蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在桥本甲状腺炎(HT组,33例)和Graves病(GD组,28例)患者甲状腺组织中的表达与分布。结果bcl鄄2蛋白表达强度GD组>C组>HT组(P<0.01);mcl鄄1蛋白表达强度GD组>C组>HT组(P<0.01);bcl鄄XL蛋白表达强度HT组和GD组强于C组(P<0.01),但HT组和GD组间差异无统计学意义(P>0.05);bax蛋白的表达强度HT组>GD组和C组(P<0.01),但GD组和C组间差异无统计学意义(P>0.05);HT组中,在淋巴细胞浸润区域附近的甲状腺滤泡上皮细胞(TEC)bcl鄄2表达弱,bax和mcl鄄1表达强;远离淋巴细胞浸润区域的TECbcl鄄2表达强,bax和mcl鄄1表达弱(P<0.05)。结论(1)抗凋亡bcl鄄2和mcl鄄1蛋白在HT中表达的减弱以及在GD中表达的增强对于HT甲状腺滤泡细胞凋亡的增加和GD甲状腺滤泡细胞的增殖可能起一定作用;(2)bax蛋白在HT中表达增强所起的促凋亡的作用对疾病的发生发展起一定作用;(3)bcl鄄2与bax表达强度的比值对于凋亡的调控起重要作用;(4)bcl鄄2家族蛋白bc     

大肠肿瘤的基因表达及与细胞凋亡的抑制关系

目的探讨 bcl-2和 P53蛋白在大肠肿瘤中的表达及与细胞凋亡关系。方法用免疫组化方法观察了45例大肠腺瘤和61例大肠癌中 bcl-2和 P53蛋白的表达。结果正常大肠粘膜中 bcl-2和 p53均未见表达,而大肠腺瘤及大肠癌阳性率均较正常明显增加(P<0.01)。大肠腺瘤 p53表达随腺瘤大小增加而增加,其中≥20 mm 组阳性率(77.8%)显著高于<10 mm 组(35.0%,P<0.05)。P53蛋白阳性率也随不典型增生程度增加而增高。p53表达与大肠癌分化程度及 Duke 分期有关。大肠癌细胞凋亡指数与 bcl-2阳性表达呈负相关。大肠腺瘤中 bcl-2和 P53蛋白的表达也呈负相关。结论 bcl-2蛋白表达对大肠癌前病变.腺瘤的增殖有一定意义,p53在大肠腺瘤癌变和大肠癌进展中起重要作用,它们是参与细胞凋亡的良好指标。     

凋亡相关基因caspase-9,bax及bcl-2在大肠癌中的表达及其意义

目的探讨凋亡相关基因easpase-9,bax和bel-2在大肠癌中的表达及其在大肠癌发生、发展中的可能作用及相互关系。方法应用免疫组化S-P法检测20例正常大肠黏膜、48例大肠腺瘤及56例大肠癌中的caspase-9、bax和bcl-2蛋白的表达。用TUNEL 法检测细胞凋亡。结果正常大肠黏膜、大肠腺瘤和大肠癌中caspase-9的阳性表达率分别为5．00％、33．33％、64．29％,其表达率在三者间差异有显著性(P<0．01)。bax在三种组织中的表达率分别为5．00％、35．42％、62．50％,三者间差异有显著性(P<0．01)。bcl-2 在三者中的阳性表达率分别为15．00％、87．50％、60．71％,三者间差异亦有显著性(P<0．01)。caspase-9,bax和bcl-2的表达与大肠癌的分化程度有关(P<0．01),与Dukes分期无关(P>05)。正常大肠黏膜、腺瘤和大肠癌中细胞凋亡指数差异有显著性(P<0．01),细胞凋亡指数与肿瘤的分化程度有关(P<0．01),与Dukes分期无关(P>0．05)。caspase-9、bax和bcl-2的表达与细胞凋亡指数有密切联系 (P<0．01)。结论肿瘤早期阶段的细胞凋亡异常,可能是大肠癌的发病原因之一。bcl-2和bax通过调节caspase-9参与大肠癌的发生。     

白藜芦醇对实验性兔骨关节炎软骨细胞凋亡及bcl-2 bax表达的影响

目的 研究白藜芦醇对兔实验性骨关节炎(OA)软骨细胞凋亡及凋亡调控基因bcl-2、bax表达的影响,探讨其治疗OA的机制.方法 30只新西兰大白兔随机平分为5组,A组(健康对照组)、B组(模型对照组)、C组(白藜芦醇高剂量干预组)、D组(白藜芦醇中剂量干预组)、E组(白藜芦醇低剂量干预组).除A组外,其他各组均以Hulth法复制膝OA模型.术后第4周开始,A组、B组每天以5 ml含0.1%二甲基亚砜的蒸馏水灌胃;白藜芦醇干预各组每天以相应剂量白藜芦醇溶液(浓度为60 mg/ml)灌胃,C、D、E组日剂量分别为120、60、30mg·kg-1·d-1,连续6周.第10周处死大白兔,取右膝关节股骨内髁内侧软骨,软骨组织切片用脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法观察软骨细胞凋亡,免疫组织化学法观察软骨细胞bcl-2和bax的表达.结果 ①模型对照组关节软骨细胞凋亡率明显高于健康对照组;白藜芦醇干预各组可不同程度地降低OA软骨细胞凋亡率,差异有统计学意义(P＜0.05),且呈剂量依赖性.②与健康对照组相比,模型对照组关节软骨细胞bcl-2和bax阳性率均升高(P＜0.01),bcl-2/bax的比值降低;白藜芦醇干预后,bcl-2阳性率升高更为明显(P＜0.01);而bax阳性率有不同程度降低(P＜0.01);bcl-2/bax的比值均不同程度提高(P＜0.01).结论 白藜芦醇通过上调bcl-2的表达,下调bax的表达,提高bcl-2/bax的比值,以抑制兔实验性OA中软骨细胞的过度凋亡,达到保护软骨,防治OA的目的 .     

β1肾上腺素能受体阻断剂与β2肾上腺素能受体激动剂联用对心力衰竭大鼠心功能及心肌细胞凋亡的影响

目的 在一定范围内激活β2肾上腺素能受体（AR）可以在不加重心室重构的前提下改善心力衰竭（心衰）大鼠心功能,推测β1AR阻断剂联合β2AR激动剂有可能进一步改善心衰大鼠心功能并减轻心肌细胞凋亡,该实验对此进行了研究并探讨了其机制。方法 随机选取9只雄性Wistar大鼠为对照组。将异丙基肾上腺素诱导的心衰大鼠随机分为美托洛尔组（n=11）,联合治疗组（n=11）,安慰剂组（n=10）。美托洛尔组给予美托洛尔50mg／kg,一日两次灌胃。联合治疗组给予非诺特罗125μg／kg,美托洛尔50mg／kg一日两次灌胃。安慰剂组给予等量生理盐水一日两次灌胃。对照组不予处理。治疗8周后应用超声心动图评价心功能,TUNEL法检测心肌细胞凋亡指数,测定Caspase-3酶活性,Westernblot测定bcl-2及bax蛋白质表达,测定脏器重量／体重,组织病理学测定胶原容积分数（CVF）。结果 （1）美托洛尔组及联合治疗组均较安慰剂组左室舒张末期直径（LVEDd）、左室收缩末期直径（LVESd）、E峰A峰比值（E／A）明显下降,短轴缩短率（FS）、射血分数（EF）则有明显增高。联合治疗组较美托洛尔组LVEDd、LVESd有进一步降低（均为P〈0．05）,FS、EF则有进一步增高（均为P〈0．01）。（2）美托洛尔组及联合治疗组较安慰剂组左室重量体重比（LVW／BW）、肺脏重量体重比（PW／BW）及CVF明显降低（均为P〈0．01）。联合治疗组LVW／BW及PW／BW较美托洛尔组进一步降低（P〈0．01）,但两组之间CVF无显著差异。（3）美托洛尔组及联合治疗组较安慰剂组心肌细胞凋亡指数（AI）及Caspase．3活性均有明显减低。联合治疗组较美托洛尔组有进一步减低（均为P〈0．01）。（4）与安慰剂组相比,美托洛尔组及联合治疗组bax蛋白表达有明显下降而bcl-2／bax显著升高,并且以联合治疗组改善更为显著（均为P〈0．01）。结论 β1AR阻断剂联合β2AR激动剂较单用β1AR阻断剂进一步改善心衰大鼠的心功能,减轻心室重构。明显降低bax蛋白表达及bcl-2／bax,减轻心肌细胞凋亡很可能是其疗效提高的机制之一。     

小鼠感染日本血吸虫后肝组织Bcl-2、Bax的表达及己酮可可碱对它的作用

目的 观察凋亡相关基因Bcl?鄄2、Bax在小鼠感染血吸虫后肝组织中的表达情况及己酮可可碱（pentoxi-fylline，PTX）对其作用。 方法 将40只小鼠随机分为4组，其中3组每只小鼠人工感染日本血吸虫尾蚴25条，1组感染后继续喂养10周，不作治疗，作为感染对照组；2组分别于感染后2周用PTX 360 mg/（kg·d）和180 mg/（kg·d）灌胃治疗8周；另1组不感染、也不接受药物治疗，与上述3组同步喂养10周，作为正常对照组。10周后将上述4组小鼠分别剖杀取肝组织，光镜观察肝组织病变；用免疫组化染色方法检测小鼠肝组织中Bcl-2、Bax的水平。 结果 感染对照组中Bcl?鄄2和Bax的表达水平较正常对照组明显增加（P＜0.05）。高剂量PTX治疗组Bcl-2水平明显高于低剂量PTX治疗组和感染对照组（P＜0.05）。而Bax的表达水平在感染对照组、低剂量PTX治疗组、高剂量PTX治疗组等3组间差异无统计学意义（P＞0.05）。同时高剂量PTX治疗组肝组织变性坏死及纤维化程度较低剂量PTX治疗组和感染对照组轻。 结论 高剂量PTX可能通过促进Bcl-2表达，减少肝细胞的变性坏死，阻断血吸虫肝纤维化的发生。     

Expression of Leukemia Inhibitory Factor in Human Pituitary Adenomas: A Morphologic and Immunocytochemical Study

Leukemia inhibitory factor (LIF) is a pleiotropic, neuropoietic cytokine found in bovine, murine, and human fetal and adult pituitary cells, mostly in corticotrophs and somatotrophs. In addition, it has been found in a few GH- and ACTH-producing human pituitary adenomas. The aim of our study was to investigate the presence of LIF in various morphologic types of human pituitary adenomas. Ninety-eight operated pituitary adenomas diagnosed by light microscopy and classified by pituitary hormone immunocytochemistry and electron microscopy were studied. Sixty-eight tumors were functioning and included 15 densely granulated (DG) and 10 sparsely granulated (SG) somatotroph (SM) adenomas, 5 lactotroph (LT), 7 mixed SM-LT and 31 corticotroph (CRT) adenomas. The remaining 30 nonfunctioning tumors included 11 gonadotroph (GON) and 19 null cell (NULL) adenomas. For immunocytochemical demonstration of LIF, a specific polyclonal antiserum was applied on formalin-fixed, paraffin-embedded tissues. The immunohistoscore ranging from 1 to 64 grades was determined by multiplying the immunostaining grade (1–4) by the staining intensity grade (1–4), and by the heterogeneity grade (1–4). Ninety adenomas (92%) were variably immunopositive for LIF. LIF was expressed in 77.5% of CRT, 81.8% of GON, 93.3% of DG-SM, and in all SG-SM, LT, SM-LT and NULL adenomas. LT adenomas showed the highest immunostaining grade, followed by SG-SM, GON, NULL, SM-LT, DG-SM, LT and CRT adenomas. GON adenomas showed the highest immunohistoscore, followed by NULL, DG-SM, SM-LT, SG-SM, LT and CRT adenomas. Nonfunctioning tumors showed a significantly higher immunohistoscore compared to functioning adenomas (p < 0.01). We conclude that LIF expression is frequent in all types of functioning and nonfunctioning pituitary adenomas.     

熊果酸对肝星状细胞增殖与凋亡的影响

目的 体外观察熊果酸对肝星状细胞增殖与凋亡的影响,探讨熊果酸诱导肝星状细胞凋亡的可能作用机制. 方法 将不同浓度熊果酸作用于肝星状细胞HSC-T6及肝细胞L02,分别在药物作用24、48、72 h后用四甲基偶氮唑盐法检测熊果酸对HSC-T6及L02细胞增殖的影响;流式细胞仪检测熊果酸对HSC-T6凋亡的影响;光学显微镜观察熊果酸作用后细胞形态学变化情况;免疫细胞化学法检测HSC-T6中Bcl-2、Bax和Caspase-3蛋白的表达情况. 结果 各种浓度的熊果酸均可抑制HSC-T6细胞的增殖,且呈剂量-时间依赖性;当熊果酸浓度为25、50、75μmol/L时可促进L02细胞增殖,浓度＞75μmol/L则表现为抑制L02细胞增殖.在病理形态学方面,熊果酸作用HSC-T6细胞48 h后,光学显微镜下可见细胞缩小变圆、核浓缩等.25、50、75 μmol/L熊果酸作用HSC-T6细胞48 h后,流式细胞仪检测显示细胞凋亡率分别为10.30%±3.85%、21.87%±4.46%、31.33%±6.18%,比对照组(2.93%±1.60%)明显升高(P＜0.01).免疫细胞化学显示Bax及Caspase-3蛋白表达较对照组升高(P＜0.05),且呈剂量依赖性,而Bcl-2蛋白表达水平与对照组无明显差异(P＞0.05).结论 在体外熊果酸可较明显地抑制HSC-T6细胞增殖,诱导其凋亡;对L02细胞的生长具有双向调节作用.熊果酸诱导HSC-T6细胞凋亡可能与降低Bcl-2/Bax比值、激活Caspase-3蛋白有关.