贝那普利对不同血糖状态糖尿病大鼠肾脏细胞凋亡及相关基因表达的影响

腹腔注射链脲佐菌素建立SD大鼠糖尿病模型，TUNEL、免疫组化检测结果显示，糖尿病持续高血糖大鼠贝那普利处理组较未处理组。肾小管凋亡细胞明显减少，Bax表达减弱，肾小球Bcl-2蛋白表达增强（均P〈0．01）；糖尿病血糖波动大鼠贝那普利处理组较未处理组。肾小管凋亡细胞数、Bax和Bcl-2蛋白表达差异均无统计学意义。     

缬沙坦对血管衰老中凋亡调控基因Bcl-2、Bax表达的影响

目的探讨缬沙坦对血管衰老中凋亡调控基因Bcl-2、Bax表达的影响。方法健康Wistar大鼠分为青年组、衰老组及缬沙坦组，测定血浆丙二醛（MDA）、超氧化物歧化酶（SOD）水平，同时采用恒速注人流体方法测定各组大鼠颈动脉血管的顺应性，利用免疫组织化学染色法、RT—PCR法和Western印迹法分析各组大鼠凋亡调控基因Bcl-2、Bax的mRNA及蛋白表达水平。结果与衰老组相比，缬沙坦组MDA浓度显著降低（P〈0．05），SOD浓度显著升高（P〈0．05），颈动脉血管的顺应性增高，其中弹性面积有显著性差异（P〈0．05），Bcl-2的mRNA及蛋白表达水平明显增高（P〈0．05），Bax的mRNA及蛋白表达水平降低（P〈0．05）。结论血管衰老有其特征性生理改变，Bcl-2、Bax的mRNA及蛋白表达的失衡可能是血管衰老的重要分子机制之一，缬沙坦有一定的逆转血管衰老的作用。     

糖尿病大鼠肾脏细胞凋亡与Bax和Bcl-2基因表达

目的　观察糖尿病大鼠肾脏细胞凋亡、Bax和 Bcl- 2表达及二者的相关性。　方法　单侧肾切除大鼠腹腔注射链脲佐菌素诱发糖尿病 ,采用原位末端标记法检测肾脏细胞凋亡 ;流式细胞术和免疫组化检测肾皮质 Bax和 Bcl- 2表达水平 ;原位杂交检测 Bax和 Bcl- 2 m RNA表达 ,并观察尿蛋白、BU N、尿肌酐等反映肾功能的有关指标。　结果　在制模后 2、4、8、12周时 ,糖尿病组大鼠较对照组肾小球、肾小管凋亡细胞数明显增多 ,Bax、Bcl- 2蛋白和 m RNA的表达显著增强 (P<0 .0 5 )。随着大鼠糖尿病病程延长 ,肾功能恶化 ,肾脏凋亡细胞数逐渐增多 ,Bax表达亦逐渐增强 ,Bax/Bcl- 2比增加 ,且肾脏凋亡细胞数与 Bax及 Bax/Bcl- 2比具有相关性 (P<0 .0 5 )。　结论　肾脏凋亡细胞的不断增加可能是糖尿病肾病发生、发展的原因之一 ,Bax和 Bcl- 2可能参与肾脏细胞凋亡的调控。     

依达拉奉对大鼠心肌再灌注抗凋亡作用的机制

目的：观察大鼠心肌缺血再灌注时依达拉奉抗凋亡作用的机制。方法：采用体外结扎大鼠左前降支制备大鼠心肌缺血再灌注模型。将Wistar大鼠随机分为对照组、缺血再灌注组、依达拉奉组（再灌注即刻经尾静脉注射依达拉奉3mg／kg）。分别用TUNEL染色法及免疫组化法检测心肌细胞凋亡、心肌凋亡蛋白Bcl-2、Bax的表达。结果：（1）凋亡检测结果示,对照组无细胞凋亡,依达拉奉组较缺血再灌注组凋亡细胞数明品减少[（13．11±1．58）比（24．33±1．38）,P〈0．01];（2）免疫组化结果示,依达拉奉组与缺血再灌注组Bcl-2、Bax蛋白表达均明显高于对照组（P〈0．01）;依达拉奉组较缺血再灌注组Bcl-2蛋白表达明显升高[（0．2300±0．0218）比（0．0924±0．0152）],而Bax蛋白表达明显降低[（0．0249±0．0042）比（O．1312±0．0173）],P均〈0．01;缺血再灌注组Bcl-2／Bax比值较对照组明显降低[（0．6906±0．0035）比（1．1632±0．0212）],依达拉奉组Bcl-2／Bax比值（7．4752±0．5573）较缺血再灌注组和对照组明显升高（P〈0．01）。结论：依达拉奉能够减少心肌细胞凋亡,心肌细胞凋亡的减轻与下调Bax蛋白表达,上调Bcl-2蛋白表达以及Bcl-2／Bax比值有关。     

甘草酸对糖尿病大鼠肾脏细胞凋亡的影响及防治糖尿病肾病的临床意义

雄性Wistar大鼠30只,随机平均分为3组,正常对照组、糖尿病组、糖尿病甘草酸（30mg／kg／d）治疗组,腹腔注射链脲佐菌素（60mg／kg）诱发糖尿病。16周后,处死大鼠,分离肾脏,观察Bcl-2、Bax的表达,取部允肾皮质电镜病理观察。结果①糖尿病组肾脏Bcl-2蛋白表达减少、Bax蛋白表达增加,甘草酸治疗组的Bcl-2蛋白表达较糖尿病组增多,而Bax蛋白表达较糖尿病组减少,电镜观察糖尿病组呈明显细胞凋亡形态学改变,甘草酸治疗组减轻。②糖尿病肾小球基底膜增厚,系膜区域扩大,甘草酸治疗组病变减轻。结论①糖尿病血糖升高可能调节Bcl-2、Bax蛋白的表达诱导细胞凋亡,而参与DN的发生与发展。②甘草酸可调查Bax、Bcl-2的表达抑制细胞凋亡,明显改善糖尿病大鼠肾脏结构与功能,延缓DN的发展。     

石斛合剂对衰老糖尿病大鼠胰腺组织凋亡相关基因Bax,Bcl-2mRNA及蛋白表达的调控

目的观察石斛合剂对衰老及衰老糖尿病（DM）大鼠胰腺组织凋亡相关基因Bax、Bcl-2mRNA及蛋白表达的影响。方法采用D-半乳糖致衰，继而加高脂高糖及注射低剂量链脲佐菌素（STZ）制备衰老DM模型，测定血清中SOD、MDA浓度，并利用RT-PCR法和Western-Blot法检测大鼠胰腺组织凋亡相关基凼Bax、Bcl-2mRNA及蛋白表达水平。结果与衰老及衰老DM组相比，石斛合剂各治疗组MDA浓度明显降低（P〈0．05），SOD浓度明显升高（P〈0．05～0．01），衰老DM石斛合剂治疗组胰腺组织BaxmRNA及蛋白水平明显降低（P〈0.05～0．01），Bcl-2mRNA及蛋白水平明显升高（P〈0．05）。结论①衰老大鼠呈现Bax／Bcl-2比值显著升高，衰老DM大鼠Bax／Bcl-2比值升高更为显著，即衰老大鼠Bax和Bcl-2mRNA及蛋白表达失衡，且这种失衡在衰老DM大鼠显得更为突出；②石斛合剂降低血糖，显著升高衰老及衰老DM大鼠SOD水平，降低MDA水平，其分子机制之一与其降低衰老DM大鼠胰腺组织Bax、增加Bcl-2 mRNA及蛋白水平，即调整Bax和Bcl-2mRNA及蛋白表达的失衡有关。     

IGF-1对MC3T3-E1细胞凋亡及凋亡调控蛋白Bax、Bcl-2表达的影响

目的 探讨胰岛素样生长因子-1（IGF-1）对小鼠前成骨样MC3T3-E1细胞凋亡基因相关蛋白Bax／Bcl-2表达的影响。方法 MC3T3-E1细胞分组培养，应用4个浓度的IGF-1（1ng／mL、10ng／mL、30ng／mL、50ng／mL）干预24h。观察MC3T3-E1细胞动态生长状况：流式细胞技术检测细胞凋亡率；免疫组织化学法检测凋亡蛋白Pax／Bcl-2的表达水平。结果 与正常血清对照组相比，无血清对照组MC3T3-E1细胞凋亡率增加（P〈0．05）；与无血清组相比，IGF-1组MC3T3-E1细胞凋亡率降低（P〈0．05），Bax表达减弱（P〈0．05），Bcl-2表达增强（P〈0．05）；各浓度组间比较，（1～50）ng／mL浓度范围内MC3T3-E1细胞凋亡率随IGF-1浓度加大而逐渐降低（P〈0．05），Bax表达逐渐减弱（P〈0．05），50ng／mL浓度组Bcl-2表达明显增强（P〈0．01），Bcl-2／Bax灰度比与细胞凋亡率呈显著正相关（r=0.917，P〈0．01）。结论 一定浓度的IGF-1同时调节Bax和Bcl-2表达，抑制无血清培养诱导的细胞凋亡。并存在剂量依赖性效应。     

川芎嗪对糖尿病鼠醛糖还原酶活性及视网膜细胞凋亡影响

雄性Wistar大鼠40只，随机平均分为4组，正常对照组、糖尿病组、糖尿病依帕司他（10mg／kg／d）治疗组、糖尿病川芎嗪（100mg／kg／d）治疗组，蝮腔注射链脲佐菌素（65mg／kg）诱发糖尿病。16周后，处死大鼠，分离晶体，测定组织AR活性，观察视网膜Bcl-2、Bax的表达。结果：1．与正常对照组相比，糖尿病组AR活性明显升高（P〈0．001），依帕司他治疗组、川芎嗪治疗组AR活性较糖尿病组明显降低（P〈0．01），而血糖无明显变化。2．糖尿病组视网膜Bcl-2、Bax蛋白表达明显增加，依帕司他治疗组、川芎嗪治疗组的Bcl-2、Bax蛋白表达较糖尿病组明显减少。结沦：1．AR过度激活通过促进Bcl～2、Bax蛋白的表达诱导细胞凋亡，而参与DR的发生与发展。2．ARts通过抑制醛糖还原酶活性，调节Bax、Bcl-2的表达抑制细胞凋亡，延缓DR的发展。3．川芎嗪对AR抑制作用与典型ARIs依帕司他相似，且价格低、副作用少，值得临床推荐应用。     

氯沙坦对糖尿病大鼠肾脏细胞凋亡及bax／bcl—2表达的影响

目的：探讨血管紧张素受体1拮抗剂（AT1Ra)对糖尿病肾脏细胞凋亡及凋亡相关基因的影响。方法：单侧肾切除大鼠腹腔注射STZ诱发糖尿病模型,每日灌胃给予AT1Ra氯沙坦（40mg/kg),共12周,采用原位末端标记法检测肾脏细胞凋亡情况,流式细胞术和免疫组化检测肾皮质Bax和Bcl-2表达,原位杂交检测bax和bcl-2 mRNA表达。结果：糖尿病组较对照组肾小球、肾小管凋亡细胞数明显增多,bax和bcl-2蛋白及mRNA表达增强,Bax/Bcl-2增高;氯沙坦治疗组较糖尿病组凋亡细胞数减少,bax表达减弱,bcl-2表达增强,Bax/Bcl-2降低。结论：氯沙坦可能通过影响凋亡相关基因bax和bcl-2表达而抑制肾脏细胞凋亡,从而发挥肾脏保护作用。     

洛沙坦对高血压大鼠血管平滑肌细胞凋亡及凋亡基因Bax和Bcl-2的影响

目的研究血管紧张素Ⅱ ATI-R阻滞剂洛沙坦对自发性高血压大鼠（SHR）血管平滑肌细胞（VSMC）凋亡及凋亡基因Bax、Bcl-2的影响。方法末端标记法（TUNEL）检测细胞凋亡及RT—PCR、免疫组化方法分别检测Bax、Bcl-2基因及其蛋白的表达。结果从（12～24）周龄，SHR VSMC凋亡逐渐减低，24周龄时，凋亡指数（APOI）低于同周龄正常血压大鼠（WKY）（P〈0.05）；洛沙坦应用8周和12周。使APOI增加71％、35％（P〈0．05）。随大鼠周龄增加，SHR胸主动脉Bax基因表达逐渐下调。与APOI变化趋势一致。洛沙坦降压治疗可使SHR胸主动脉Bax蛋白表达上调。Bcl-2蛋白下调。结论洛沙坦长期降压治疗促进Bax蛋白表达和／或抑制Bcl-2蛋白表达，其机制可促进VSMC凋亡。     

Classification and characterization of hereditary types 2A, 2B, 2C, 2D, 2E, 2M, 2N, and 2U (unclassifiable) von Willebrand disease.

All variants of type 2 von Willebrand disease (VWD) patients, except 2N, show a defective von Willebrand factor (VWF) protein (on cross immunoelectrophoresis or multimeric analysis), decreased ratios for VWF:RCo/Ag and VWF:CB/Ag and prolonged bleeding time. The bleeding time is normal and FVIII:C levels are clearly lower than VWF:Ag in type 2N VWD. High resolution multimeric analysis of VWF in plasma demonstrates that proteolysis of VWF is increased in type 2A and 2B VWD with increased triplet structure of each visuable band (not present in types 2M and 2U), and that proteolysis of VWF is minimal in type 2C, 2D, and 2E variants that show aberrant multimeric structure of individual oligomers. VWD 2B differs from 2A by normal VWF in platelets, and increased ristocetine-induced platelet aggregation (RIPA). RIPA, which very likely reflects the VWF content of platelets, is normal in mild, decreased in moderate, and absent in severe type 2A VWD. RIPA is decreased or absent in 2M, 2U, 2C, and 2D, variable in 2E, and normal in 2N. VWD 2M is usually mild and characterized by decreased VWF:RCo and RIPA, a normal or near normal VWF multimeric pattern in a low resolution agarose gel. VWD 2A-like or unclassifiable (2U) is distinct from 2A and 2B and typically featured by low VWF:RCo and RIPA with the relative lack of high large VWF multimers. VWD type 2C is recessive and shows a characteristic multimeric pattern with a lack of high molecular weight multimers, the presence of one single-banded multimers instead of triplets caused by homozygosity or double hereozygosity for a mutation in the multimerization part of VWF gene. Autosomal dominant type 2D is rare and characterized by the lack of high molecular weight multimers and the presence of a characteristic intervening subband between individual oligimers due to mutation in the dimerization part of the VWF gene. In VWD type 2E, the large VWF multimers are missing and the pattern of the individual multimers shows only one clearly identifiable band, and there is no intervening band and no marked increase in the smallest oligomer. 2E appears to be less well defined, is usually autosomal dominant, and accounts for about one third of patients with 2A in a large cohort of VWD patients.     

Structure-function of Hb Marseille-Long Island [alpha 2 beta 2 N-methionyl-2(NA2) His----Pro

Suspensions of red cells containing Hb Marseille-Long Island showed decreased oxygen affinity and low interaction with 2,3-diphosphoglycerate. Oxygen equilibrium studies of the purified component confirmed these abnormalities. Oxidation rate measurements of carbonmonoxy-Hb Marseille and carbonmonoxy-Hb A by ferricyanide showed an increased rate for the former, suggesting an increased dissociation constant for carbon monoxide. Nuclear Magnetic Resonance spectra in the high field region revealed small changes in the proximal region of the heme pocket. These results indicated that the mutation causes a perturbation at a distance from the mutation site.     

Different RBC aggregating properties of the Aalpha2Bbeta2gammaA2 and Aalpha2Bbeta2gammaAgamma' subpopulations of human fibrinogen

Human fibrinogen (TF) has been separated into two fractions: F1 - homodimers with respect to the gamma chain, and F2 - heterodimers composed of gammaA and gamma' polypeptides. Their rouleaux-inducing properties were as follows: (1) both, at the same concentration of 0.8%, were less effective than TF; (2) F1 produced larger rouleaux even under static conditions of a hemocytometer where F2 was silent; (3) F2 induced the process when a suspension was gently sheared between microscopic slides. Since the synthetic peptide gamma'(414-427) inhibited the rouleau formation in a mixture with F2, the C-terminal amino acids of the gamma' polypeptide probably bind the molecule to the cell. The inhibition was feebly visible in the native ratio of F1/F2, implicating a compensatory effect of F1.     

Identification of SH2B2beta as an inhibitor for SH2B1- and SH2B2alpha-promoted Janus kinase-2 activation and insulin signaling

The SH2B family has three members (SH2B1, SH2B2, and SH2B3) that contain conserved dimerization (DD), pleckstrin homology, and SH2 domains. The DD domain mediates the formation of homo- and heterodimers between members of the SH2B family. The SH2 domain of SH2B1 (previously named SH2-B) or SH2B2 (previously named APS) binds to phosphorylated tyrosines in a variety of tyrosine kinases, including Janus kinase-2 (JAK2) and the insulin receptor, thereby promoting the activation of JAK2 or the insulin receptor, respectively. JAK2 binds to various members of the cytokine receptor family, including receptors for GH and leptin, to mediate cytokine responses. In mice, SH2B1 regulates energy and glucose homeostasis by enhancing leptin and insulin sensitivity. In this work, we identify SH2B2beta as a new isoform of SH2B2 (designated as SH2B2alpha) derived from the SH2B2 gene by alternative mRNA splicing. SH2B2beta has a DD and pleckstrin homology domain but lacks a SH2 domain. SH2B2beta bound to both SH2B1 and SH2B2alpha, as demonstrated by both the interaction of glutathione S-transferase-SH2B2beta fusion protein with SH2B1 or SH2B2alpha in vitro and coimmunoprecipitation of SH2B2beta with SH2B1 or SH2B2alpha in intact cells. SH2B2beta markedly attenuated the ability of SH2B1 to promote JAK2 activation and subsequent tyrosine phosphorylation of insulin receptor substrate-1 by JAK2. SH2B2beta also significantly inhibited SH2B1- or SH2B2alpha-promoted insulin signaling, including insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1. These data suggest that SH2B2beta is an endogenous inhibitor of SH2B1 and/or SH2B2alpha, negatively regulating insulin signaling and/or JAK2-mediated cellular responses.