中药对Fas/FasL细胞凋亡系统的作用

Fas及其配体FasL是近年来研究得最为深入的有关细胞凋亡的膜表面分子,而中药因其作用广,疗效好,毒副作用小而成为目前药物研究最为热门的领域之一。现选择中药对Fas/FasL细胞凋亡系统作用进行了综述,希望能因此为中药影响细胞凋亡机制作为疗效的中药新药研发提供参考。     

Fas/FasL对糖尿病心肌病的影响

糖尿病心肌病(DCM)是指排除了高血压性心脏病、冠状动脉粥样硬化心脏病、心脏瓣膜病及其他心脏病变所致的心肌损伤后诊断的一种特异性心肌病变。心肌细胞凋亡为其重要的发病机制,晚期主要表现为心室重塑和心脏功能障碍。 Fas/FasL是重要的凋亡信号通路,主要是Fas/FasL为主的膜受体通路作用于细胞凋亡过程,在糖尿病的形成过程中起有重要作用,从而对糖尿病心肌病产生影响。本文阐述了Fas/FasL在DCM病理、生理中的作用,以及对未来治疗DCM的展望。     

BPA-induced apoptosis of rat Sertoli cells through Fas/FasL and JNKs/p38 MAPK pathways

Bisphenol-A was examined for its effects on cultured Sertoli cells established from 18 to 22-day-old rat testes. Results indicated that exposure to BPA (0, 30, 50 and 70 μM) decreased the cell viability in a concentration-dependent manner and induced cell apoptosis. Apoptosis-caused cell death was observed in cells exposed to 50 and 70 μM BPA. The mRNA expressions of Fas, FasL and caspase-3 were all elevated, and the protein expressions of FasL and cleaved caspase-3 were also increased. In addition, levels of phosphorylation of JNKs/p38 MAPK were also increased and then activated JNKs/p38 MAPK up regulated target gene expressions, such as c-jun and CHOP. Translocation of NF-κB into nuclei indicated the activation of NF-κB after treatment with BPA. Taken together, observed results suggest that BPA induces apoptosis of Sertoli cells by the activation of JNKs/p38 MPAK and translocation of NF-κB, and Fas/FasL system plays a critical role in the initiation of apoptosis.     

Intervention of selenium on apoptosis and Fas/FasL expressions in the liver of fluoride-exposed rats

Fluorosis is a major public health problem in numerous areas around the world, including China. To alleviate this problem, selenium has been used. In this study, we aimed to investigate the influence of selenium on apoptosis in fluorosis-affected rat livers and determine the optimal selenium concentration in drinking water to fight fluorosis. The protein levels of Fas in NaF and NaF + Se (0.375 and 0.75 mg/L) groups as well as FasL in NaF, Se (0.75 and 1.5 mg/L), and NaF + Se (0.375 mg/L) groups were significantly increased compared with those in the control group. The mRNA levels of Fas in NaF and Se (1.5 mg/L) groups as well as FasL in NaF and NaF + Se (0.375 mg/L) groups were significantly increased. The protein levels of Fas in NaF + Se (1.5 mg/L) group and FasL in three NaF + Se groups were significantly decreased compared with those in the NaF group. The mRNA levels of Fas in the three NaF + Se groups and FasL in NaF + Se (0.75 and 1.5 mg/L) groups were significantly decreased. Compared with the control group, activity of GSH-Px, and SOD in the NaF group decreased obviously and MDA content increased obviously; activity of SOD in 1.5 mg/L Se group decreased obviously. Compared with the NaF group, activity of GSH-Px in NaF + Se (1.5 mg/L) group significantly increased, and MDA content decreased obviously. Thus, fluoride induced apoptosis in the liver, thereby causing liver damage in the rats. Selenium could alleviate fluorosis-induced liver injury. In particular, selenium at 1.5 mg/L is considered the optimum concentration against fluorosis.     

环孢素 A对心肌缺血再灌注 Fas／Fas L蛋白表达及心肌细胞凋亡的影响

目的：探讨环孢素A （CSA ）对心肌缺血再灌注大鼠Fas／FasL蛋白表达的影响及CSA对心肌细胞的保护作用。方法选取30只SD大鼠随机分为假性手术组（S组）、缺血在灌注组（IR组）、CSA预处理组（CSA组）,每组各10只,CSA组给予每天10 mg／kg的CSA与腹腔内注射,10 d后建立大鼠心肌缺血再灌注模型,IR组及CSA组结扎左冠状动脉前降肢30 min后复灌注3 h ,S组仅行穿线处理,不进行结扎。分别于结扎前（T0）、再灌注即刻（T1）、30 min（T2）、60 min（T3）、90 min（T4）、120 min（T5）采用ELISA测定各组大鼠血清超敏肌钙蛋白I（cT‐nI）、肌酸激酶同工酶（CK‐MB）水平。于试验结束后采用流式细胞仪测定各组大鼠血清心肌细胞凋亡率、钙调神经磷酸酶（CaN）活性,采用免疫组织化学法测定Fas／FasL蛋白水平。结果与T0,IR组、CSA组T1～ T5时段cTnI、CK‐MB水平显著升高（ P＜0．05）。与 S 组相比,IR 组、CSA 组 T1～ T5时段 cTnI、CK‐MB 水平显著升高（P＜0．05）。CSA组T1～T5时段cTnI、CK‐MB水平显著低于IR组（P＜0．05）。与IR组,CSA组心肌缺血再灌注后心肌细胞凋亡率、CaN活性及Fas／FasL蛋白水平显著下降（P＜0．05）。结论 CSA预处理心肌缺血再灌注大鼠可通过抑制CaN信号降低大鼠心肌梗死面积,同时能有效减少Fas／FasL蛋白表达,延缓心肌细胞缺血再灌注损伤,具有保护心肌细胞作用。     

神经节苷脂钠联合亚低温治疗重度颅脑损伤对神经细胞Fas/FasL信号通路以及细胞凋亡的影响

目的　探究与分析神经节苷脂钠联合亚低温治疗重度颅脑损伤对神经细胞Fas/FasL信号通路以及细胞凋亡的影响。方法　选取晋城市人民医院2020年2月至2022年2月收治的重度颅脑损伤患者118例,按照不同治疗方案分为对照组与观察组,每组59例,其中对照组有7例,观察组有5例患者因死亡、中途退出研究者导致临床资料缺失,最终对照组有52例,观察组有54例纳入到最后研究。对照组中男30例,女22例,年龄（35.69±3.14）岁;观察组中男28例,女26例,年龄为（35.77±3.20）岁。对照组给予常规降低颅内压、保护脑细胞、抗感染以及解痉挛等治疗,观察组在其基础上给予神经节苷脂钠联合亚低温治疗,对比两组临床疗效、治疗前后格拉斯哥昏迷评分法（GCS）评分、大脑中动脉（MCA）血流速度及脑脊液中肿瘤坏死因子-α（TNF-α）、Fas、Fas配体（FasL）、半胱天冬氨酸酶-9（Caspase-9）蛋白水平和住院时间。采用χ²检验、t检验、F检验。结果　对照组总有效率为82.69%（43/52）、住院时间为（44.05±13.52）d,观察组总有效率为96.30%（52/54）、住院时间为（35.36±14.11）d,观察组与对照组相比临床总有效率较高,住院时间较短,差异均有统计学意义（χ²=5.271,P=0.022,t=3.236,P=0.002）。治疗后1周、2周、4周对照组GCS评分分别为（6.96±0.87）分、（7.54±1.02）分、（8.98±1.12）分,观察组分别为（7.43±0.86）分、（8.58±0.99）分、（10.58±1.24）分,两组治疗后1周、2周、4周分别与治疗前相比GCS较高,观察组治疗后1周、2周及4周分别与对照组治疗后1周、2周及4周相比,GCS较高,差异均有统计学意义（t=2.796,P=0.006;t=5.324,P<0.001;t=6.977,P<0.001）。治疗后1周、2周、4周对照组MCA分别为（88.47±7.58）cm/s、（81.98±12.84）cm/s、（72.87±12.84）cm/s,观察组分别为（85.33±8.10）cm/s、（75.45±14.15）cm/s、（66.86±13.78）cm/s,两组治疗后1周、2周、4周分别与治疗前相比MCA血流速度较低,观察组治疗后1周、2周、4周分别与对照组治疗后1周、2周、4周相比,MCA血流速度较低,差异均有统计学意义（t=2.062,P=0.042;t=2.490,P=0.014;t=2.234,P=0.022）。治疗后2周、4周对照组的TNF-α、Fas、FasL及Caspase-9分别为（1.04±0.51）mg/L、（0.79±0.32）mg/L,（34.55±7.25）μg/L、（27.10±5.58）μg/L,（89.34±5.77）μg/L、（20.87±6.55）μg/L,（23.54±5.47）pmol/L、（14.23±4.69）pmol/L;观察组分别为（0.83±0.41）mg/L、（0.36±0.12）mg/L,（40.14±8.20）μg/L、（10.35±4.14）μg/L,（102.47±5.78）μg/L、（17.53±5.28）μg/L,（40.69±6.78）pmol/L、（8.69±0.25）pmol/L,两组治疗后4周与治疗后2周相比,TNF-α、Fas、FasL及Caspase-9较低,观察组治疗后2周与对照组治疗后2周相比,TNF-α较低,Fas、FasL及Caspase-9较高,观察组治疗后4周与对照组治疗后4周相比,TNF-α、Fas、FasL及Caspase-9较低,差异均有统计学意义（均P<0.05）。结论　神经节苷脂钠联合亚低温治疗重度颅脑损伤可改善患者的昏迷程度,临床效果突出,通过作用于神经Fas/FasL信号通路的过程,对细胞凋亡的发生产生抑制效果,缩短住院时间,获得良好预后。     

Doxorubicin-induces NFAT/Fas/FasL cardiac apoptosis in rats through activation of calcineurin and P38 MAPK and inhibition of mTOR signalling pathways

This study investigated the role of NFAT/Fas/FasL axis in cardiomyocyte apoptosis following doxorubicin (DOX) treatment in rats and evaluated the involvement and regulation of all NFAT members in cardiac apoptosis. Forty adult male Wistar rats were divided equally into control or DOX-treated groups (15 mg/kg over 2 weeks). Cardiomyocytes were cultured and pre-incubated with various inhibitors and activators (10 μmol/L) prior to DOX exposure (1 μmol/L). In the left ventricles and cultured cells, DOX increased cytoplasmic protein levels of cytochrome C, Bax and increased the activities of caspase-8, caspase3, ERK1/2, JNK, and P38 mitogen-activated protein kinases (MAPKs), reducing levels of Bcl-2 and the activity of mTOR, and inducing cell death. In addition, DOX enhanced mRNA and protein levels of Fas and FasL. Furthermore, the nuclear and cytoplasmic levels of NFAT1 and nuclear accumulation of NFAT2-4were increased with DOX treatment. The inhibition of calcineurin with FK506 significantly inhibited the nuclear levels of NFAT2 and NFAT4 and the inhibition of P38 MAPK with SB203580 inhibited the nuclear and cytoplasmic accumulation of NFAT1. However, the activation of mTOR by IGF-1 significantly lowered NFAT3. In conclusion, NFAT/Fas/FasL-induced cell death in cardiac myocytes of DOX-treated rats is regulated, at least, by the activation of calcineurin and P38 MAPK and inhibition of mTOR.     

p38MAPK介导的Fas/FasL凋亡通路在缺血性脑损伤中的作用研究

目的探讨p38MAPK介导的Fas/FasL凋亡通路在大鼠缺血性脑损伤中的作用。方法①制作大鼠全脑缺血模型,免疫印迹法检测假手术组、脑缺血复灌6 h、12 h、1 d、3 d组p-p38MAPK、p38MAPK蛋白表达。②免疫印迹法检测假手术组、缺血复灌组、溶剂对照组和SB203580组p-p38MAPK、p38MAPK、FasL、Fas和Caspase-3蛋白表达。结果①与假手术组相比,脑缺血再灌注6 h、12 h、1 d、3 d组p-p38MAPK蛋白表达水平逐渐升高,于1 d达高峰(P均<0.05)。②与缺血复灌组和溶剂对照组相比,SB203580组p-p38MAPK、FasL和Caspase-3表达水平显著降低(P均<0.05)。结论 p38MAPK介导的Fas/FasL凋亡通路在缺血性脑损伤中发挥了重要作用。     

p,p′-DDE induces testicular apoptosis in prepubertal rats via the Fas/FasL pathway

1,1-Dichloro-2,2 bis(p-chlorophenyl) ethylene (p,p′-DDE), the major metabolite of 2,2-bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT), is a known persistent organic pollutant and male reproductive toxicant. It has antiandrogenic effect. However, the mechanism by which p,p′-DDE exposure causes male reproductive toxicity remains unknown. To elucidate the mechanism underpinning the testicular effects of p,p′-DDE, we sought to investigate Fas/FasL apoptotic pathway in the testis of prepubertal rats, including Fas, FasL, caspase-8, -3, and NF-κB. Animals were administered with different doses of p,p′-DDE (0, 20, 60, 100 mg/kg b.wt) every other day by intraperitoneal injection for 10 days. The results indicated that p,p′-DDE exposure at over 20 mg/kg b.wt showed the induction of apoptotic cell death. p,p′-DDE could induce increase in the MDA level, and decrease in SOD and GSH-Px activity. Significant elevations in the mRNA levels of Fas along with an increase in FasL, caspase-3, -8 were observed in 100 mg/kg b.wt group. In protein level, p,p′-DDE could induce increase of FasL and reduction of procaspase-8. NF-κB p65 was activated by p,p′-DDE treatment in rat testis. In addition, the activities of caspase-3, -8 were increased in 100 mg/kg b.wt group. Taken together, these results lead us to speculate that in vivo exposure to p,p′-DDE might induce testicular apoptosis in prepubertal rats through the Fas/FasL pathway.     

Celastrol induces apoptosis in non-small-cell lung cancer A549 cells through activation of mitochondria- and Fas/FasL-mediated pathways

Celastrol is a natural compound extracted from the traditional Chinese medicinal herb, Tripterygium wilfordii Hook. It has attracted interests for its potential anti-inflammatory and antitumor effects. However, the molecular mechanisms of celastrol-induced apoptosis in cancer cells remain unclear. In this study, we investigated the effects of celastrol on the human non-small-cell lung cancer (NSCLC) cell line A549 in vitro. Celastrol caused a dose- and time-dependent growth inhibition of A549 cells with an IC₅₀ of 2.12 μM at 48 h treatment. Celastrol induced A549 cells apoptosis as confirmed by annexin V/propidium iodide staining and DNA fragmentation. Celastrol-induced apoptosis was characterized by cleavage of caspase-9, caspase-8, caspase-3, and PARP protein, increased Fas and FasL expression, and a reduction in the mitochondrial membrane potential. Furthermore, celastrol induced the release of cytochrome c. Celastrol also up-regulated the expression of pro-apoptotic Bax, down-regulated anti-apoptotic Bcl-2, and inhibited Akt phosphorylation. These results demonstrate that celastrol can induce apoptosis of human NSCLC A549 cells through activation of both mitochondria- and FasL-mediated pathways.     

诊断超声照射后人早孕绒毛细胞Fasm RNA FasL mRNA表达的改变

目的　探讨诊断超声与人早孕绒毛Fas/FasLmRNA表达的关系。方法　对拟进行人工流产的 2 4例早孕 (4 5～ 6 0d)妇女随机分为 4组 :Ⅰ组 (对照组 )、Ⅱ组 (10min组 )、Ⅲ组 (2 0min组 )和Ⅳ组 (30min组 )。应用HP 85 0 0彩色多普勒超声诊断仪 ,对孕囊进行不同时间的持续照射 ,照射后 2 4h取材。用原位杂交技术检测上述各组绒毛滋养层细胞Fas/FasLmRNA表达情况。结果　超声照射后 10min组绒毛滋养层细胞Fas/FasLmRNA表达率与对照组差异无显著性 (P >0 0 5 ) ,2 0min组和 30min组滋养层细胞表达率明显增加 ,显著大于对照组和10min组 (P <0 0 0 1)。结论　诊断超声持续照射早孕孕囊组织 >10min可引起绒毛滋养层细胞Fas/FasLmRNA表达率增加 ,从而诱导滋养层细胞凋亡增加     

地塞米松对SLE患者外周血淋巴细胞凋亡及Fas、FasL表达的影响

目的观察活动期系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL)凋亡状态及Fas、FasL的表达,探讨地塞米松(Dex)对其的影响。方法分离19例SLE患者及10例健康人PBL,植物血凝素(PHA)刺激培养72h,流式细胞仪检测其凋亡率及Fas、FasL的表达。SLE患者的PBL,再经Dex作用后检测上述指标。结果SLE患者PBL在PHA刺激培养72h后凋亡率(42.81±7.52)%,较健康对照组(4.74±0.59)%明显升高(P<0.01),Fas表达SLE组(47.67±4.80)MFI,较健康对照组(21.70±5.16)MFI增高(P<0.01),FasL表达阳性细胞百分率与健康对照无明显差别(2.48±0.24)%,(2.64±0.31)%(P>0.05);SLE患者PBL经Dex培养后凋亡率(63.92±7.58)%较未处理组增高;Fas表达、FasL阳性细胞百分率两组间差异无统计学意义。结论SLE患者存在PBL凋亡及Fas、FasL表达的异常;Dex可加速SLE患者PBL的凋亡,且可能是通过Fas/FasL以外的途径实现。     

Role of the Fas/Fas Ligand death receptor pathway in ginseng saponin metabolite-induced apoptosis in hepg2 cells

This research team found in previous studies, that the ginseng saponin metabolite IH901 induces apoptosis in HepG2 cells via a mitochondrial-mediated pathway, which resulted in the activation of caspase-9 and subsequently of caspase-3 and -8. Based on these results, the involvement of the Fas/Fas ligand (FasL) death-receptor pathway, in IH901-induced apoptosis in HepG2 cells, was investigated. Levels of Fas and the Fas ligand (FasL) mRNA or protein were not increased by IH901, rather they were decreased significantly at 18 h post treatment. Soluble FasL (sFasL) was detectable by immunoprecipitation analysis in the medium of HepG2 cells treated with IH901. Increased levels of sFasL were inversely correlated with the levels of FasL. Preincubation of HepG2 cells with antagonistic anti-Fas antibody showed little protective effect, if any, on IH901-induced cell death. At a 30 microM (24 and 48 h) and 40 microM (24 h) concentration of IH901, the cytotoxic effect of IH901 was less then 50%, anti-Fas antibody prevented IH901-induced cell death. However, at a 60 microM (24 and 48 h) and 40 microM (48 h) concentration of IH901, cell death rates were about 80% or more and most of the chemopreventive and chemotherapeutic effects of IH901 were manifested. Blocking the Fas receptor did not influence IH901-induced cell death. These results indicate that the Fas/FasL system is engaged, but not required for IH901-induced cell death, at pharmacologically significant concentrations.     

Tetrazolium violet induces apoptosis via caspases-8, -9 activation and Fas/FasL up-regulation in Rat C6 glioma cells

Tetrazolium violet (TV), a tetrazolium salt, was synthesized as a novel and potent anticancer agent with a broad spectrum of anticancer activity against many cancer cells. A previous study showed that tetrazolium violet inhibited cell growth, and induced cell cycle arrest and apoptosis in C6 Rat glioma cells. It also showed that treatment of cells with TV for 24 h resulted in a dramatic up-regulation of p53, and an increase in the activity of caspase-3, accompanied with a significant increase of Bax/Bcl-2 ratio. In this study, we further investigated which Fas/FasL and caspase were activated by TV during the apoptosis. Annexin-V-propidium iodide (PI) binding assay and nucleosome ELISA assay further indicated that TV induced a typical apoptosis, in a time-dose-dependent manner. The data showed that the activity of Fas/FasL and caspase-8 and -9 were significantly enhanced by the compound, which suggested that TV might be used as a Fas/FasL and caspases promoter to initiate brain cancer cell apoptosis.     

TFPI-2的生物学作用及其与肿瘤的关系

组织因子途径抑制物-2(tissue factor pathway inhibitor-2,TFPI-2)属于Kuniz型丝氨酸蛋白酶抑制物家族的一员。近来研究发现,组织因子途径抑制物-2在多种肿瘤增殖、侵袭和转移过程中发挥明显的抑制作用,然而目前其作用机制尚不明确。该文就组织因子途径抑制物-2的生物学作用及其与肿瘤的关系方面做一总结。     

不同年龄白内障晶状体上皮细胞中caspase-3、Fas/FasL的表达及意义

目的：探讨不同年龄白内障晶状体上皮细胞（LECs）中caspase-3、Fas/FasL的表达及临床意义。方法选择2007年9月~2012年2月本院眼科收治的白内障患者50例为研究对象,其中25～49岁设为白内障中青年组,≥50岁设为白内障老年组,各25例,并收集20例成人尸体解剖非白内障眼球为对照组,其中25～49岁设为中青年对照组,≥50岁设为老年对照组,各10例。采用免疫组织化学法检测各组晶状体前囊膜组织中caspase-3、Fas、FasL的表达。结果白内障老年组、白内障中青年组、老年对照组和中青年对照组LECs中凋亡细胞百分率分别为34.0%、16.0%、2.0%、3.0%,白内障老年组显著高于白内障中青年和老年对照组,白内障中青年组显著高于中青年对照组,差异有统计学意义（P＜0.01）。白内障老年组LECs中caspase-3、Fas、FasL蛋白的阳性表达率分别为84.0%、60.0%、68.0%,白内障中青年组LECs中caspase-3、Fas、FasL蛋白的阳性表达率分别为44.0%、24.0%、32.0%,老年对照组LECs中caspase-3、Fas、FasL蛋白的阳性表达率分别为10.0%、10.0%、20.0%,中青年对照组LECs中caspase-3、Fas、FasL蛋白的阳性表达率分别为10.0%、0.0%、0.0%,4组caspase-3、Fas、FasL蛋白阳性表达率比较,差异有统计学意义（P＜0.01）。老年白内障患者和中青年白内障患者中,LECs凋亡与caspase-3和Fas、FasL表达水平均呈正相关（r=0.621、0.583、0.621,P＜0.01）。结论老年性白内障形成过程可能与LECs的凋亡及caspase-3、Fas/FasL的表达调控有关。     

Fas／FasL基因在11酸睾酮诱导恒河猴无精子症或少精子症中的作用

目的：研究凋亡相关基因Fas/FasL在11酸睾酮诱导无精子症或少精子症的作用。方法：TUNEL用于检测睾丸细胞的凋亡信号。免疫组化和Western blot用于对这些凋亡相关基因所表达的蛋白进行定量或定性分析。结果：11酸睾酮处理后,生精细胞的凋亡信号和Fas/FasL蛋白的表达呈现时间依赖性方式增强,至第30天达到最大值。结论：Fas系统可能参与启动和调节11酸睾酮诱导的生精细胞的凋亡。     

清热化瘀滋阴方对系统性红斑狼疮外周血淋巴细胞凋亡及Fas、FasL基因表达的影响

目的：探讨中药清热化瘀滋阴方对系统性红斑狼疮（SLE）患者外周血淋巴细胞凋亡及Fas、FasL基因表达的影响。方法：临床筛选SLE患者80例,完全随机分为治疗组（清热化瘀滋阴方配合激素组）和对照组（单用激素组）各40例,服药3个月,淋巴细胞分离液分离治疗前后SLE患者外周血淋巴细胞,采用流式细胞仪检测淋巴细胞凋亡率;采用RT-PCR法检测淋巴细胞Fas、FasL基因表达情况。结果：清热化瘀滋阴方治疗后SLE患者外周血淋巴细胞凋亡率较治疗前明显下降（P〈0.01）,并明显低于对照组（P〈0.01）;两组治疗后Fas、FasL基因的表达均明显下降,治疗组Fas基因的表达明显低于对照组（P〈0.01）。结论：清热化瘀滋阴方可能通过降低SLE患者外周血淋巴细胞凋亡率,下调淋巴细胞Fas、FasL基因的表达水平,改善免疫紊乱而发挥作用。     

5-氟尿嘧啶诱导Jurkat细胞凋亡及Fas、FasL的表达

目的探讨5-氟尿嘧啶（5-FU）诱导Jurkat细胞株的凋亡和Fas、FasL的表达。方法不同浓度的5-FU分别处理Jurkat细胞12h和24h后,流式细胞仪检测细胞表达CD95、CD95L的百分率。结果与对照组相比,5-FU在10、20、40μg.mL-1浓度下明显促进Jurkat细胞表达Fas、FasL;且5-FU在24h比处理12h有更强的诱导效果。结论 5-FU可显著提高Jurkat细胞凋亡率,并增强Fas、FasL的表达率。