补肾健脾方干预去势大鼠骨骼肌caspase-3和caspase-8的表达

背景：骨骼肌与骨质疏松症关系密切,导致其衰退的主要原因是细胞的凋亡,这种凋亡可能是Caspase家族蛋白所介导的。目的：探讨补肾健脾方对去势大鼠骨骼肌caspase-3和caspase-8调控作用。方法：SD大鼠48只等随机分为对照组、模型组、中药组、西药组,后3组大鼠摘除双侧卵巢建立骨质疏松模型,对照组仅切除周围少量脂肪组织。造模2周后,中药组给予补肾健脾方（2.979g/kg）灌胃,西药组给予戊酸雌二醇片（0.104mg/kg）灌胃,模型组和对照组给予蒸馏水灌胃,1次/d。结果与结论：①干预12周后,双能X线骨密度仪测量发现,相比于对照组,模型组大鼠左侧股骨的骨密度值和骨矿含量均明显降低（P〈0.01）;而中药组和西药组的骨密度值均高于模型组（P〈0.05）,且2组骨密度值和骨矿含量接近（P〉0.05）。②酶标免疫分析显示,与对照组相比,模型组骨骼肌中Caspase-3和Caspase-8表达水平明显上升（P〈0.05）。中药能显著降低大鼠骨骼肌中Caspase-3的表达水平（P〈0.05）,而西药能显著减低Caspase-8表达水平（P〈0.05）。③说明补肾健脾方对卵巢切除所致的骨质疏松症有明显的治疗作用,能明显提高骨密度,而且能显著减低Caspase-3的水平,但不能显著减低Caspase-8水平,显示补肾健脾方不是通过死亡受体途径去抑制细胞凋亡的。     

补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用

目的:探讨补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用机制。方法:将新西兰大白兔9只随机分为中药组、西药组、空白组,每组3只。三组分别用中药提取液、雌二醇混悬液按中药剂量和同体积(约10 ml)蒸馏水灌胃,1个月后抽取含药血浆。选用1 d龄SD大鼠获取原代的成骨细胞体外培养,分为中药组、西药组、凋亡空白组、正常空白组。中药组、西药组分别给予对应的含药血浆培养;凋亡空白组和正常空白组均给予常规血清培养。24 h后检测ASK1、ERO1和Caspase-12含量。结果:在ASK1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P0.05);且西药组比中药组更能显著降低ASK1m RNA的表达,其差异有统计学意义(P0.05)。在ERO1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P0.05),且西药组比中药组更能显著降低ERO1蛋白的表达,其差异有统计学意义(P0.05)。在Caspase-12方面,中药组、西药组与凋亡空白组相比,差异无统计学意义(P0.05)。结论:补肾健脾中药复方通过ASK1、ERO1所介导的方式对成骨细胞的凋亡进行调控。     

补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用*

目的：探讨补肾健脾中药复方对成骨细胞中ASK1、ERO1、Caspase-12调节作用机制。方法：将新西兰大白兔9只随机分为中药组、西药组、空白组三组,每组3只。三组分别用中药提取液、雌二醇混悬液按中药剂量和同体积(约10ml)蒸馏水灌胃,1月后抽取含药血浆。选用1日龄SD大鼠获取原代的成骨细胞体外培养,分为中药组、西药组、凋亡空白组、正常空白组。中药组、西药组分别给予对应的含药血浆培养；凋亡空白组和正常空白组均给予常规血清培养。24小时后采用检测ASK1、 ERO1和Caspase-12的含量。结果：在ASK1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P<0.05)；而且西药组比中药组更能显著降低ASK1 mRNA的表达,其差异有统计学意义(P<0.05)。在ERO1方面,中药组、西药组与凋亡空白组相比,差异有统计学意义(P<0.05),而且西药组比中药组更能显著降低ERO1蛋白的表达,其差异有统计学意义(P<0.05)。在Caspase-12方面,中药组、西药组与凋亡空白组相比,差异无统计学意义(P>0.05)。结论：补肾健脾中药复方通过ASK1、ERO1所介导的方式对成骨细胞的凋亡进行调控。     

黄芩素对阿霉素肾病大鼠肾小管上皮细胞凋亡的调节作用

目的:探讨黄芩素对阿霉素肾病大鼠肾小管上皮细胞凋亡的调节作用。方法:18只雄性Wistar大鼠随机分为正常对照组(正常组,n=6)、阿霉素肾病组(模型组,n=6)和黄芩素治疗组(治疗组,n=6),模型组和治疗组一次性尾静脉注射阿霉素2.0mg/kg诱导阿霉素肾病动物模型,正常组注射等量生理盐水。造模后第1天开始,治疗组予黄芩素300 mg/(kg·d)灌胃,正常组和模型组予等量蒸馏水灌胃,共治疗8周。8周时留取各组大鼠24 h尿液标本及肾脏组织标本,使用TUNEL检测肾小管上皮细胞凋亡,免疫组化检测肾组织中caspase-3表达,western Blot方法检测肾皮质中Bcl-2及caspase-3蛋白水平。结果:与正常组比较,模型组24 h尿蛋白定量明显升高(P0.05),肾小管上皮细胞凋亡数量明显增加(P0.05),肾皮质中caspase-3蛋白水平明显增加(P0.05),Bcl-2蛋白水平明显下降(P0.05)。与模型组比较,治疗组24 h尿蛋白定量明显下降(P0.05),TUNEL阳性肾小管上皮细胞数量明显减少(P0.05),肾皮质中caspase-3蛋白水平明显下降(P0.05),Bcl-2蛋白水平明显增加(P0.05)。结论:黄芩素对阿霉素肾病具有一定的保护作用,其作用机制可能与调节肾小管上皮细胞凋亡有关。     

瑞舒伐他汀对急性心肌梗死大鼠心肌细胞凋亡的影响

目的:探讨急性心肌梗死(AMI)大鼠心肌细胞凋亡和心肌细胞凋亡基因Fas及Caspase-3表达变化及瑞舒伐他汀对其的影响。方法:结扎30只雄性SD大鼠左冠状动脉前降支造成AMI模型,存活的24只随机分为两组:他汀组(12只)和对照组(12只),另取8只雄性SD大鼠作为假手术组。术后第1天起他汀组给予瑞舒伐他汀1mg/(kg·d)灌胃,对照组、假手术组给予等量生理盐水灌胃。4周后,比较各组大鼠心肌细胞凋亡,凋亡基因Fas及Caspase-3表达变化。转移酶介导脱氧尿苷三磷酸缺口末端标记法检测心肌凋亡细胞。免疫组织化学法检测Fas、Caspase-3蛋白水平。反转录-聚合酶链反应法分析Fas、Caspase-3基因mRNA表达水平。结果:他汀组和对照组心肌细胞凋亡指数,Fas、Caspase-3蛋白阳性染色细胞指数及Fas、Caspase-3基因的mRNA表达水平较假手术组明显增加(P<0.05),而他汀组较对照组减少(P<0.05)。结论:AMI大鼠心肌细胞凋亡指数、心肌组织Fas、Caspase-3蛋白及Fas、Caspase-3基因的mRNA表达水平明显增高。瑞舒伐他汀对AMI大鼠的心肌细胞凋亡、细胞凋亡基因fas及Caspase-3表达水平增高有较好的拮抗作用。     

去卵巢牙周炎模型大鼠牙槽骨吸收与补骨脂的干预

背景:补骨脂在治疗骨质疏松症方面有明显效果.目的:观察补骨脂对去卵巢牙周炎大鼠牙槽骨代谢、牙槽骨密度及高度的影响.方法:将30只雌性Wistar 大鼠随机等分为3 组:去卵巢组和补骨脂组行双侧卵巢切除术,假手术组仅手术不切除卵巢,补骨脂组于卵巢切除第2 天灌胃3 g/kg 补骨脂水剂,1次/d.去卵巢2周,所有大鼠采用结扎上颌磨牙的方法建立牙周炎1540007,模型结果与结论:上颌磨牙结扎12 周,与假手术组比较,去卵巢组大鼠血清雌二醇、钙离子浓度显著降低(P < 0.05),碱性磷酸酶水平显著升高(P < 0.05),X 射线片及苏木精-伊红染色显示去卵巢组大鼠上颌骨骨密度及高度显著降低(P < 0.05) ;与去卵巢组比较,补骨脂组大鼠血清钙离子浓度显著升高(P < 0.05),碱性磷酸酶水平显著降低(P < 0.05),上颌骨骨密度及高度显著增高(P < 0.05),且与假手术组比较差异无显著性意义(P > 0.05).说明雌激素缺乏促进实验性牙周炎大鼠牙槽骨吸收,补骨脂可有效阻止雌激素降低导致的牙槽骨高度、骨密度降低.     

大剂量高压氧对脑梗死大鼠细胞色素C及caspase-3的影响

目的观察单次高压氧(HBO)治疗9 h对永久性大脑中动脉阻塞(MCAO)大鼠细胞色素C和caspase-3水平的影响。方法48只Sprague-Dawley大鼠制作永久性MCAO模型,分为对照组(n=24)和高压氧组(n=24),高压氧组大鼠于造模成功后3 h予单次高压氧治疗9 h,压力0.2 MPa。于造模成功后3 h、13 h、24 h行大鼠Garcia神经功能评分,于造模成功后13 h、24 h TUNEL法检测大鼠脑组织细胞凋亡数,ELISA法检测缺血半暗带脑组织细胞色素C及活性caspase-3水平。结果两组大鼠造模成功后13h、24 h神经功能均有明显改善,两组间无显著性差异(t2.07,P0.05)。两组造模成功后13 h、24 h均见凋亡细胞,高压氧组明显少于对照组(t6.57,P0.01)。高压氧组细胞色素C和caspase-3的水平在造模成功后24 h低于对照组(t2.41,P0.05)。结论大剂量高压氧超早期治疗可降低脑缺血后细胞色素C和caspase-3水平,减少细胞凋亡,可能是高压氧治疗脑梗死的机制之一。     

生长激素对大鼠心肌缺血/再灌注后心肌细胞凋亡及凋亡相关基因Caspase-3 Cleaved caspase-3蛋白表达的影响

目的 研究生长激素(Growth hormone, GH)对大鼠心肌缺血/再灌注(ischemia reperfusion,I/R) 后心肌细胞凋亡及相关基因Caspase-3、Cleaved caspase-3蛋白表达的影响.方法 24只大鼠随机分为三组,假手术组(sham-operated group,n=8,仅手术24 h)、心肌缺血/再灌注(I/R)组(n=8)、GH组(n=8),后两组均缺血30 min,再灌注24 h,GH组于建立模型前7 d每天给予人重组生长激素(reconbination human growth hormone,rhGH)针剂皮下注射(1 U/kg体质量),其他二组等量生理盐水注射.以TUNEL法检测心肌细胞凋亡情况, DAB免疫组化法检测心肌细胞Caspase-3、Cleaved caspase-3蛋白表达并进行心肌组织病理学检查.结果 大鼠心肌I/R 24 h后心肌细胞凋亡指数明显上升(对照于假手术组,P<0.05),心肌细胞Caspase-3、Cleaved caspase-3蛋白表达呈阳性染色指数明显升高(P<0.05),心肌病理检查见心肌缺血区呈大小不一的坏死灶,缺血心肌间有炎症细胞浸润,心肌排列不整齐(HE染色),而GH组心肌细胞凋亡率及心肌细胞Caspase-3、Cleaved caspase-3蛋白表达呈阳性染色指数明显好于I/R组(P<0.05),心肌细胞炎症细胞也明显减少,坏死灶也少于I/R组.结论 GH可以减少心肌I/R后心肌细胞凋亡及细胞Caspase-3、Cleaved caspase-3染色阳性指数,说明GH对I/R后的心肌具有保护作用.     

阿托伐他汀对大鼠心肌急性缺血细胞凋亡及caspase-3基因表达影响

目的:观察不同剂量阿托伐他汀对大鼠急性缺血心肌细胞凋亡及caspase-3基因表达的影响。方法:取雄性SD大鼠79只,随机分为模型组、阿托伐他汀高剂量组(20 mg/(kg.d))、中剂量组(10 mg/(kg.d))、低剂量组(2mg/(kg.d))和生理盐水对照组。除模型组只开胸外,其余大鼠术前连续灌胃3 d,第4天结扎冠状动脉左前降支造成急性心肌梗死模型。伊文氏蓝和TTC染色法确定缺血和梗死心肌范围。原位末端标记检测细胞凋亡,RT-PCR检测caspase-3 mRNA的表达。结果:与对照组比较,阿托伐他汀高?中剂量组可明显减少心肌梗死面积,明显减少心肌细胞凋亡和心肌内caspase-3 mRNA基因表达。不同剂量组间差异无统计学意义(P0.05)。结论:高、中剂量阿托伐他汀可减少大鼠急性缺血心肌细胞损伤,与能减少心肌细胞凋亡及抑制caspase-3 mRNA基因表达有关。     

补肾通络方对SD大鼠骨性关节炎、骨质疏松症及骨性关节炎并骨质疏松症模型骨密度的影响

背景:临床上发现通过补肾通络方治疗骨性关节炎及骨质疏松症有良好的临床疗效.目的:尝试建立骨性关节炎+骨质疏松症动物模型,观察补肾通络方对SD大鼠骨性关节炎、骨质疏松症及骨性关节炎+骨质疏松症模型骨密度的影响.设计、时间及地点:随机对照观察实验,于2008-08/2009-01在新疆医科大学科研中心完成.材料:4月龄雌性SD大鼠70只,随机分为正常对照组、骨性关节炎组、骨性关节炎+补肾通络方组、骨质疏松症组、骨质疏松症+补肾通络方组、骨性关节炎+骨质疏松症组、骨性关节炎+骨质疏松症+补肾通络方组,每组10只.补肾通络方由骨碎补、杜仲、山萸肉、熟地黄、补骨脂、秦艽、寄生、独活、桃仁、红花、当归、川芎、白芍等组成.方法:取20只大鼠,采用改良Hulth法制作骨性关节炎模型,选择10只于术后1个月灌胃给予补肾通络方;取20只大鼠,采用去势法制作骨质疏松症模型,选择10只于术后1个月灌胃给予补肾通络方;取20只大鼠按上述方式同时制作骨性关节炎、骨质疏松症模型,选择10只于术后1个月灌胃给予补肾通络方.按大鼠体质量灌胃给药,根据人鼠换算出服药量为10.31 mg/(kg·d),2次/d,连续4周.正常对照组不做任何处理.主要观察指标:电镜、光镜观察大鼠左侧股骨内髁关节软骨组织形态学变化,双能X射线观察大鼠左侧股骨骨密度变化.结果:光镜下观察3组模型大鼠骨小梁局部区域变细,排列较紊乱,纵向结构受到破坏,连续性较差,出现较多的断裂点,关节软骨变薄、退变;电镜下观察3组模型大鼠模型组大鼠关节软骨细胞异形性增多,表现为细胞核形态不规则,细胞壁断裂不完整,细胞器减少,细胞核固缩,染色质分布不均匀,并有大量软骨细胞凋亡.骨性关节炎组、骨质疏松症组、骨性关节炎+骨质疏松症组大鼠骨密度较正常对照组明显下降(P<0.05),经补肾通络方干预后骨密度上升(P<0.05).结论:补肾通络方对骨性关节炎、骨质疏松症SD大鼠骨密度有上调作用,但其有效作用机制尚需进一步研究探讨.     

运动在防治骨质疏松症中的应用

运动疗法在骨质疏松症非药物干预中的有益作用及其机制,运动方式和运动强度对骨质疏松症的防治作用,根据对骨质疏松症危险因素的评估、骨密度测定和运动能力的评定,将骨质疏松症分为3组。针对3组的特点制定不同的运动疗法方案。     

Activation of caspase-3 is an initial step triggering accelerated muscle proteolysis in catabolic conditions

With trauma, sepsis, cancer, or uremia, animals or patients experience accelerated degradation of muscle protein in the ATP-ubiquitin-proteasome (Ub-P'some) system. The initial step in myofibrillar proteolysis is unknown because this proteolytic system does not break down actomyosin complexes or myofibrils, even though it degrades monomeric actin or myosin. Since cytokines or insulin resistance are common in catabolic states and will activate caspases, we examined whether caspase-3 would break down actomyosin. We found that recombinant caspase-3 cleaves actomyosin, producing a characteristic, approximately 14-kDa actin fragment and other proteins that are degraded by the Ub-P'some. In fact, limited actomyosin cleavage by caspase-3 yields a 125% increase in protein degradation by the Ub-P'some system. Serum deprivation of L6 muscle cells stimulates actin cleavage and proteolysis; insulin blocks these responses by a mechanism requiring PI3K. Cleaved actin fragments are present in muscles of rats with muscle atrophy from diabetes or chronic uremia. Accumulation of actin fragments and the rate of proteolysis in muscle stimulated by diabetes are suppressed by a caspase-3 inhibitor. Thus, in catabolic conditions, an initial step resulting in loss of muscle protein is activation of caspase-3, yielding proteins that are degraded by the Ub-P'some system. Therapeutic strategies could be designed to prevent these events.     

半胱氨酸天冬氨酸蛋白酶3在急性脑及脊髓损伤修复中的作用

目的探讨半胱氨酸天冬氨酸蛋白酶3的分子生物学研究进展及它在中枢神经系统疾病中的作用.资料来源应用计算机检索Medline数据库1995-01/2005-02相关文章,检索词为"caspase-3","apoptosis","center rnerve system","emergency diseases"分别组合进行检索,限定文章语言种类为English.资料选择对资料进行初审,选取与半胱氨酸天冬氨酸蛋白酶3有关的分子生物学方面的文章和半胱氨酸天冬氨酸蛋白酶3与中枢神经系统疾病有关的随机对照实验.资料提炼共收集到156篇关于半胱氨酸天冬氨酸蛋白酶3与中枢神经系统有直接关系的文章,其中有25篇符合标准,排除131篇.排除的131篇系重复或者与本文关系不大.资料综合对检索到的文章的相关信息综合加以概括综述,总结出在急性脑损伤、急性脊髓损伤、脑缺血、脑出血后存在半胱氨酸天冬氨酸蛋白酶3活性增高,应用广谱半胱氨酸天冬氨酸蛋白酶抑制剂Z-VAD-fmk可显著降低半胱氨酸天冬氨酸蛋白酶3活性.结论半胱氨酸天冬氨酸蛋白酶3是凋亡过程中最重要的蛋白酶,是多种凋亡途径的共同下游效应部分,是细胞凋亡蛋白酶级联反应的必经之路.半胱氨酸天冬氨酸蛋白酶3激活后可裂解细胞内固有的和保护性酶类,进而形成细胞凋亡的形态学及生物化学特征性改变.DNA片段化是急性脑损伤早期神经细胞凋亡的重要生化特征,DNA片段化的形成是必需的.     

Unshackling caspase-7 for cancer therapy

Numerous solid tumors and hematologic malignancies acquire resistance to apoptosis-inducing chemotherapeutic drugs by downregulating the key effector caspase-3. These cells rely on caspase-7 to execute the apoptotic program, yet binding with XIAP constitutively inhibits active caspase-7 (p19/p12-CASP7). In this issue, Lin et al. describe how a newly synthesized drug is able to disrupt the XIAP:p19/p12-CASP7 complex and induce apoptosis in caspase-3–deficient cancer cells in vitro and in vivo. As this compound appears to exhibit minimal toxicity on normal tissues, it may represent a promising therapeutic agent to help treat caspase-3–deficient tumors.     

p38-MAPK signals survival by phosphorylation of caspase-8 and caspase-3 in human neutrophils

Neutrophil apoptosis occurs both in the bloodstream and in the tissue and is considered essential for the resolution of an inflammatory process. Here, we show that p38-mitogen-activated protein kinase (MAPK) associates to caspase-8 and caspase-3 during neutrophil apoptosis and that p38-MAPK activity, previously shown to be a survival signal in these primary cells, correlates with the levels of caspase-8 and caspase-3 phosphorylation. In in vitro experiments, immunoprecipitated active p38-MAPK phosphorylated and inhibited the activity of the active p20 subunits of caspase-8 and caspase-3. Phosphopeptide mapping revealed that these phosphorylations occurred on serine-364 and serine-150, respectively. Introduction of mutated (S150A), but not wild-type, TAT-tagged caspase-3 into primary neutrophils made the Fas-induced apoptotic response insensitive to p38-MAPK inhibition. Consequently, p38-MAPK can directly phosphorylate and inhibit the activities of caspase-8 and caspase-3 and thereby hinder neutrophil apoptosis, and, in so doing, regulate the inflammatory response.     

Muscle-specific expression of IGF-1 blocks angiotensin II-induced skeletal muscle wasting

Advanced congestive heart failure is associated with activation of the renin-angiotensin system and skeletal muscle wasting. We previously showed that angiotensin II infusion in rats produces cachexia secondarily to increased muscle proteolysis and also decreases levels of circulating and skeletal muscle IGF-1. Here we show that angiotensin II markedly downregulates phospho-Akt and activates caspase-3 in skeletal muscle, leading to actin cleavage, an important component of muscle proteolysis, and to increased apoptosis. These changes are blocked by muscle-specific expression of IGF-1, likely via the Akt/mTOR/p70S6K signaling pathway. We also demonstrate that mRNA levels of the ubiquitin ligases atrogin-1 and muscle ring finger-1 are upregulated in angiotensin II-infused WT, but not in IGF-1-transgenic, mice. These findings strongly suggest that angiotensin II downregulation of IGF-1 in skeletal muscle is causally related to angiotensin II-induced wasting. Because the renin-angiotensin system is activated in many catabolic conditions, our findings have broad implications for understanding mechanisms of skeletal muscle wasting and provide a rationale for new therapeutic approaches.     

Src inhibition enhances paclitaxel cytotoxicity in ovarian cancer cells by caspase-9-independent activation of caspase-3

Src tyrosine kinase has been found to be overexpressed and activated in a high proportion of ovarian cancers and ovarian cancer cell lines. Furthermore, Src activation is associated with activation of growth and survival signaling pathways. The present study was conducted in order to determine the effects of Src inhibition on ovarian cancer cell survival in response to chemotherapeutic agents. Inhibition of Src, either pharmacologically or through expression of a Src dominant-negative fusion construct, enhanced the cytotoxicity of two different classes of chemotherapeutics: paclitaxel and cisplatinum, in both mouse and human ovarian cancer cells. Interestingly, Src inhibition also restored sensitivity to drug-resistant ovarian cancer cells. The increased cytotoxicity in response to Src inhibition was associated with a large increase in processing and activation of caspase-3. The activation of caspase-3 seems to be independent of cytochrome c release and caspase-9 activation. The present study indicates that Src tyrosine kinase may provide an important target for small molecule inhibition in ovarian cancer.