Effects of rhein on human articular chondrocytes in alginate beads

This study was designed to investigate the effects of rhein, the active metabolite of diacerhein, on the metabolic functions of human chondrocytes cultured in alginate beads.Enzymatically isolated osteoarthritic (OA) chondrocytes were cultured in alginate beads in a well-defined culture medium for 12 days. Rhein was tested in a range of concentrations comprised between 10(-7) and 4 x 10(-5)M, in the presence or absence of 10(-10)M IL-1beta. Interleukin (IL)-6 and -8, macrophage inflammatory protein (MIP-1beta), stromelysin-1 (MMP-3), aggrecan (AGG), tissue inhibitor of metalloproteinases-1 (TIMP-1), prostaglandin E(2) (PGE(2)) and nitric oxide (NO) productions were assayed. Cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) mRNA steady-state levels were also quantified. In the basal condition, 10(-5)M rhein increased by 46.5% the production of AGG, decreased by 17-30% the production of IL-6, MMP-3, NO and MIP-1beta but enhanced by 50% the production of PGE(2). IL-1beta increased IL-6, IL-8, MIP-1beta, NO, PGE(2) and MMP-3 productions, but inhibited AGG and TIMP-1 synthesis. Rhein partially reversed the effect of IL-1beta on TIMP-1 and NO production, had no effect on AGG, IL-6 and MIP-1beta production, but up-regulated the IL-1beta stimulated PGE(2) production. The COX-2 and iNOS mRNA levels and IL-8 production were not modified by rhein.Overall, these results contribute to explain the clinical efficiency of rhein and give new information on its mechanisms of action.     

男性高尿酸血症患者血尿酸与PAI-1和内皮素的关系

目的：探讨中老年男性高尿酸血症患者血浆纤溶酶原激活物抑制剂-1（PAI-1）与内皮素（ET）水平变化，观察高尿酸对血管内皮细胞功能的影响。方法：随机选择35岁以上的男性高尿酸血症患者138例（高尿酸血症，UA组），年龄、地域相匹配的健康者80例为正常对照组。采用酶联免疫吸附双抗体夹心法测定PAI-1，采用放射免疫法测定ET。胰岛素抵抗指数采用稳态模型评估（HOMA—IR）。采用尿酸氧化酶法测定血尿酸。结果：（1）UA组PAI-1、ET水平高于正常对照（均P〈0．01）。（2）UA组收缩压、舒张压、腰围与臀围比值、体质量指数与正常对照组相比差异有统计学意义（P〈0．05或P〈0．01）。（3）uA组胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-c）、空腹血糖、糖负荷后2h血糖、空腹胰岛素、HOMA—IR均显著高于正常对照组（P〈0．05或P〈0．01）。（4）UA组高密度脂蛋白胆固醇（HDL-c）低于正常对照组（P〈0．05）。（5）在以PAI-1为因变量的逐步回归分析中血尿酸、腰围与臀围比值、HOMA—IR进入回归方程（调整后R2=0．77，P〈0．01）。（6）在以ET为因变量的逐步回归分析中，血尿酸、血肌酐、腰围与臀围比值、HOMA—IR进入回归方程（调整后R2=0．70，P〈0．01）。结论：高尿酸是血管内皮细胞功能障碍独立的危险因素，降低血尿酸水平有望改善血管内皮细胞功能，减少动脉粥样硬化的发生和发展。     

柚皮苷激活Maxi K对糖尿病心脏的保护作用

目的探讨柚皮苷(naringin, NA)通过激活大电导钙激活钾离子通道(the large conductance Ca;activated K;channels, Maxi K)对糖尿病心肌病的保护作用。方法高脂饲料喂养SD大鼠,随后腹腔注射链脲佐菌素(streptozotocin, STZ)建立糖尿病大鼠模型。造模后随机分为模型组(DCM),柚皮苷治疗组(NA),柚皮苷+Maxi K特异性抑制剂治疗组(NA+PAX),每组8只大鼠。治疗组大鼠连续给药12周,定期检测血糖。结束后观察大鼠心功能、形态及纤维化改变;并检测心脏Maxi K的α及β亚基变化。结果超声显示NA可部分恢复大鼠心功能,而特异性阻断Maxi K后,NA对心脏的保护作用明显下降;纤维化分析显示NA治疗后可降低大鼠胶原蛋白及纤连蛋白表达,该作用可被PAX部分逆转;而Western blot结果显示Maxi Kα及β亚基在DCM组表达下降,NA治疗后无明显改变。结论柚皮苷通过促进细胞膜表面Maxi K通道开放而非增加其表达产生对糖尿病大鼠的心脏保护作用。     

基于脑源性神经营养因子信号通路探讨针刺干预抑郁症的作用机制

通过检索相关数据库,对近年来针刺通过调控脑源性神经营养因子（BDNF）介导的信号通路及作用机制来干预抑郁的基础研究进行总结,发现针刺主要通过调节BDNF/酪氨酸激酶受体B(TrKB)、组织纤溶酶激活因子(tPA)/BDNF信号通路,调控突触可塑性、BDNF表观遗传等发挥抗抑郁作用。但当前大部分研究缺乏对BDNF通路进行反向验证,并且对通路或作用机制的研究仅限于经典的上下游靶点,同时局部的穴位与BDNF通路和疾病靶器官之间的特异性机制有待进一步阐明。     

低分子肝素和尿激酶对大鼠肾小球炎症保护作用的比较

目的比较低分子量肝素和尿激酶对肾小球炎症反应的保护作用。方法3月龄雌性Wistar大鼠40只,随机分为对照组(NC组)、脂多糖(LPS)组(L组)、LPS 氨甲环酸组(LT组)、LPS 氨甲环酸 低分子量肝素组(LTH组)及LPS 氨甲环酸 尿激酶组(LTU组),每组8只。采用直接免疫荧光检测肾小球纤维蛋白沉积和CD11b阳性细胞分布情况,Western blotting检测内皮细胞细胞间黏附分子-1(ICAM-1)蛋白质表达。结果NC组大鼠肾小球内无纤维蛋白沉积和CD11b阳性细胞;ICAM-1有少量表达。与NC组比较,L组纤维蛋白沉积(9.1%±1.6%)和CD11b阳性细胞(11.2±2.1)增多(P<0.05);ICAM-1表达(0.23±0.09)明显上调(P<0.05)。与L组比较,LT组纤维蛋白沉积(23.4%±3.2%)和CD11b阳性细胞(20.4±3.5)进一步增多;ICAM-1表达(0.44±0.16)进一步上调(P<0.05)。与LT组比较,LTH组和LTU组纤维蛋白沉积(分别为11.3%±1.4%,10.6%±1.5%)和CD11b阳性细胞(分别为7.3±1.8,8.4±1.6)明显减少(P<0.05或0.01);ICAM-1表达(分别为0.19±0.10,0.18±0.09)明显下调(P<0.05),LTH、LTU两组间无统计学差异。结论低分子肝素与尿激酶均能有效减少纤维蛋白沉积,起到减轻肾小球炎症反应的作用。     

基于STAT3通路探讨补肾活血方抑制骨细胞凋亡

目的探讨补肾活血方通过信号转导与转录激活子3(STAT3)通路抑制骨细胞的凋亡对骨质疏松的治疗作用。方法选取SPF级健康雌性C57BL/6小鼠75只,随机分为正常对照组11只及造模组64只。造模组采用去势法建立骨质疏松症模型,正常对照组仅暴露双侧卵巢而不切除。将造模成功的小鼠随机分为模型组、补肾活血方低、中、高剂量组及阳性对照组,各12只。模型组和正常对照组分别灌胃给予蒸馏水5 mL·kg^-1,1次/d;补肾活血方低、中、高剂量组分别灌胃给予1.79、3.57、7.14 g·mL^-1补肾活血方药液5 mL·kg^-1,1次/d;阳性对照组灌胃给予0.14 mg·mL^-1尼尔雌醇混悬液5 mL·kg^-1,1次/d。各组小鼠均治疗12周。检测比较各组骨形态参数,骨细胞凋亡指数(AI),以及骨组织B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Janus激酶2(JAK2)、STAT3表达水平。结果与模型组比较,各组小鼠骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、...     

AP-1活化在高糖诱导肾小管上皮细胞转分化中的作用

目的探讨转录活化蛋白-1（activator protein-1,AP-1）在高糖诱导的肾小管上皮细胞表型转化中的作用。方法将体外培养的人肾小管上皮细胞（human kidney proximal tubular epithelial cell,HK2）分为正常对照组、高糖组、AP-1阻断组。光镜、透射电镜观察细胞形态的改变,免疫细胞化学法检测α-平滑肌肌动蛋白（α—SMA）和细胞角蛋白（cytokeratin,CK）的表达,用凝胶电泳迁移率改变分析法（electrophoretic mobility shift assay,EMSA）检测HK2细胞AP-1的改变,RT—PCR法检测CKmRNA的表达,Westernblot检测α-SMA蛋白的表达。结果高糖作用48h后,HK2细胞由椭圆形贴壁生长变为长梭形,胞体拉长,透射电镜下,细胞表面微绒毛明显减少,胞质内粗面内质网丰富,而AP-1阻断组较高糖组明显减轻;EMSA结果分析表明,在高糖刺激下,AP-1结合活力较正常对照组增加79．22％（P〈0．05）,而AP-1阻断剂可明显抑制AP—1的活化,较高糖组降低51．19％（P〈0．05）;免疫细胞化学和RT—PCR检测显示,高糖组和AP-1阻断组CKmR—NA和蛋白水平明显降低（P〈0．05）,而AP—1阻断组显著高于高糖组（P〈0．05）;免疫细胞化学和Westernblot检测显示,高糖组和AP-1阻断组α—SMA蛋白表达明显高于正常组（P〈0．05）,而AP-1阻断组显著低于高糖组（P〈0．05）。结论高糖能够导致肾小管上皮细胞AP-1活化,诱导其表型转化。     

颞下颌关节紊乱病关节液中尿纤溶酶原激活物及其受体的表达

目的 检测颞下颌关节(TMJ)关节液中尿纤溶酶原激活物(urokinase-type plasminogen activator,uPA)及受体(urokinase-type plasminogen activator receptor,uPAR)的分泌量,探讨TMJ液中uPA及uPAR与颞下颌关节紊乱病(TMD)的关系.方法 采用酶联免疫吸附实验法检测56例TMD患者的64侧关节和10名健康志愿者的16侧关节的关节液标本中的uPA及uPAR的量.将符合纳入标准的48侧TMD患者的关节液标本根据临床诊断分为关节炎性组(A组)、结构紊乱组(B组)、骨关节病组(C组),每组16侧;10名健康志愿者的16侧关节液设为对照组(D组).结果 TMD中A组、B组、C组、D组uPA的检出量分别为(51.200±8.786)ng/L、(53.667±11.894)ng/L、(81.278±25.828)ng/L、(17.960±9.859)ng/L;uPAR检出量分别为(5.840±0.179)ng/L、(6.168±1.465)ng/L、(2.416±0.525)ng/L、(2.416±0.525)ng/L.uPA和uPAR表达量均高于健康对照组(P＜0.05),C组uPA和uPAR表达量高于A组及B组(P＜0.05),但A组与B组差异无统计学意义(P＞0.05).结论 TMJ内过度分泌的uPA和uPAR可能参与了TMD关节组织的病理破坏过程;关节液中uPA和uPAR的水平可部分反映TMD病变的程度,可作为反映TMD关节损害和代谢的客观生化分子标志.     

Angiotensin converting enzyme DD genotype affects the changes of plasma plasminogen activator inhibitor-1 activity after primary percutaneous transluminal coronary angioplasty in acute myocardial infarction patients

Angiotensin converting enzyme (ACE) DD genotype, and plasminogen activator inhibitor (PAI-1) 4G/4G genotype have been reported to affect PAI-1 activity in control subjects and atherosclerotic patients, but no data are available on the influence of angiotensin II type 1 receptor (AT1R) A1166C polymorphism on the inhibitor levels. The degree of fibrinolytic activation after percutaneous transluminal coronary angioplasty (PTCA) has been found to affect the risk of restenosis. The aim of this study was to investigate the possible influence of ACE I/D, AT1R A1166C, and PAI-1 4G/5G polymorphisms on the changes of PAI-1 activity after primary successful percutaneous transluminal angioplasty. In 29 consecutive acute myocardial infarction patients, undergoing primary successful angioplasty, genotyping of ACE I/D, AT1R A1166C, and PAI-1 4G/5G polymorphisms was performed by polymerase chain reaction and restriction fragment length polymorphism analysis, and PAI-1 plasma activity (chromogenic method) was assessed before and after angioplasty. Following angioplasty, PAI-1 activity increased in 10 of 29 patients and decreased or remained unchanged in 19 of 29. ACE DD genotype was significantly (P = 0.04) associated with an increase of PAI-1 activity post angioplasty (OR DD/ID+II = 6.5, CI 95% 4.83-8.22). Whereas no effect of PAI-1 4G/5G and AT1R A1166C polymorphisms on PAI-1 response to angioplasty was demonstrated, these data suggest that renin-angiotensin system genes are involved in the regulation of the fibrinolytic response to balloon injury, possibly affecting angiotensin converting enzyme activity. This interaction between the renin-angiotensin system and hemostasis may be a mechanism by which ACE DD genotype affects the risk of restenosis after percutaneous transluminal angioplasty.     

在放射性肾损伤纤维性修复中的分子病理机制

目的为探讨纤溶酶原激活物抑制因子-1(PAI-1)在放射性肾损伤纤维性修复中的作用和意义,揭示其发病的分子病理机制。方法24只雄性健康的Wistar大鼠以30Gy60Coγ线单次双肾局部照射,于照射后4个不同时相点活杀,取肾组织进行常规病理学观察及原位分子杂交,观察PAI-1mRNA表达的变化。结果纤维化的肾组织PAI-1表达较正常对照组增高,与纤维化程度呈正相关。结论PAI-1可能在肾放射性损伤后纤维性修复的发生及发展中起重要的作用。