丹芍化纤胶囊对体外活化HSCs增殖、凋亡及Fas/FasL途径的影响

目的:研究中药复方制剂丹芍化纤胶囊对体外活化肝星状细胞(HSCs)增殖、凋亡及Fas蛋白、caspase-3 mRNA表达的影响,探讨该药抗肝纤维化的作用机理.方法:制备丹芍化纤胶囊大鼠药物血清,分离培养大鼠HSCs,并分为小牛血清组、正常大鼠血清组、丹芍化纤药物血清组3组,分别加入含5%、10%、20%以上3种血清的DMEM培养基干预培养的HSCs,然后测定以下指标:MTT比色法测定原代HSCs增殖情况,TUNEL法测定传一代HSCs凋亡情况,免疫细胞化学SABC法测定传一代HSCs内Fas蛋白表达情况,荧光实时定量RT-PCR法检测传一代HSCs内caspase-3 mRNA表达情况.结果:丹芍化纤药物血清组较同浓度小牛血清组及正常大鼠血清组明显抑制HSCs增殖,促进HSCs凋亡,且活化的HSCs内Fas蛋白表达显著增多、增强,caspase-3 mRNA含量亦明显增加.同时,药物血清浓度越高,以上作用越明显,呈剂量依赖性.结论:丹芍化纤胶囊能显著抑制HSCs增殖,并通过增加活化HSCs内Fas蛋白及caspase-3 mRNA表达(即激活HSCs内Fas/FasL途径)诱导该细胞凋亡.     

晚癌康2号对原发性肝癌细胞凋亡基因Bcl-2 Bax和Fas表达的影响

目的:探讨中药晚癌康2号治疗原发性肝癌的作用机制.方法:选择生长良好的肝癌细胞株,分别培养实验组和对照组的肿瘤细胞,滴加稀释度为1:50抗人Bcl-2、Bax和Fas及PCNA的单克隆抗体,采用荧光免疫组化显示技术和激光共聚焦显微细胞仪的定量技术(Lsser Scan Confobal microscope,LSCM)观测扫描细胞的荧光值.结果:晚癌康2号能影响人体肝癌细胞凋亡相关基因Bcl-2、Fas表达,使Bcl-2表达减少,Fas表达增加,对Bax表达无影响.结论:晚癌康2号对人体原发性肝癌细胞有治疗作用,其作用机制可能是通过对凋亡相关基因调控的表达来实现的.     

小柴胡汤冲剂治疗子宫内膜异位症的作用机制

目的探讨小柴胡汤冲剂治疗大鼠子宫内膜异位症(内异症)的作用机制.方法建立大鼠内异症动物模型,利用免疫组化的方法,观察分析各用药组在位、异位内膜Fas及Caspase-3蛋白的表达.结果小柴胡汤治疗组内异症大鼠异位内膜Fas蛋白、Caspase-3蛋白的表达水平明显高于其在位内膜,而在对照组中的表达却恰恰相反,丹那唑组中两者相差不明显.结论小柴胡汤冲剂治疗大鼠内异症的作用机制可能是通过上调Fas蛋白的表达,促进异位内膜细胞的凋亡而实现的.     

Fas 、FasL 在胃癌发生发展中的表达及其意义

 目的 研究Fas、FasL在胃癌发生发展中的表达，并探讨其临床病理意义。方法 应用免疫组化SABC法检测60例胃癌组织、癌旁交界组织、距肿瘤边缘5cm组织及20例良性胃疾病组织和20例正常胃黏膜中Fas、FasL的表达。结果 胃癌组织、癌旁交界组织、距肿瘤边缘5cm组织和良性胃疾病、正常胃黏膜组织的Fas阳性表达率分别为46．6％、51．6％、83．3％、85％、95％；FasL的阳性率则分别为86．6％、81．6％、26．6％、20％、10％。在胃癌不同分化程度之间，Fas和FasL表达有显著性差异(P<0．05)。而胃癌不同TNM分期，有无淋巴结转移之间则无显著性差异(P>0．05)。结论 Fas、FasL的表达失衡与胃癌的发生发展有着密切关系，并与胃癌的分化和免疫逃逸有关。     

葛根素注射液对急性脑缺血模型大鼠脑细胞损伤后Fas蛋白表达的影响

目的:探讨葛根素注射液对急性脑缺血模型大鼠脑细胞损伤后Fas表达的影响.方法:采用闭夹大脑中动脉造成大鼠局部急性脑缺血模型,用免疫组化SP法测定Fas表达情况,光镜观察脑组织损伤程度.结果:葛根素组死亡细胞数显著少于对照组(P＜0.01),葛根素组与对照组比较Fas阳性细胞数差异无显著意义(P＞0.05),各组Fas阳性细胞数与死亡细胞数间无明显相关性(P＞0.05).结论:葛根素对急性脑缺血模型大鼠脑细胞损伤具有保护作用,其机制可能主要通过上调HSP70蛋白的表达而实现,而与Fas蛋白的表达及细胞凋亡等相关不强.     

Pentamer peptide from Fas antigen ligand inhibits tumor‐growth with solid‐bound form found by peptide array

Abstract: Despite their low molecular weight and simple structure, peptides regulate vital reactions of various cells. With higher standards of safety and purity required in clinical researches, chemically synthesized peptides are considered as an ideal alternative material for animal‐derived materials in the regulation of cellular events. However, effective high‐throughput assay for studying peptide‐cell interactions has not been established to design peptide with objective function. Here, we report the effectiveness of the utilization of peptide array combined with cell assay for the design of cell‐interactive peptides. As a model case, peptide that regulates tumor cell viability with support‐bound form was explored. By culturing cells on peptide array, we found a novel 5‐mer sequence CNNLP (Cys‐Asp‐Asp‐Leu‐Pro) that strongly inhibits viability of tumor cells (leukemia and adenocarcinoma) from human Fas antigen ligand. We here indicate the advantageous features of peptide array, which are ‘feasibility in examining combinatorial amino acid substitutions’ and ‘indicative data consist of effective and ineffective substitutions from an assay’, contributes greatly for studying peptide‐cell interaction. These features differentiate peptide array from other peptide library screening strategy. As a result, we succeeded in obtaining 29 novel tumor‐growth inhibitory peptides with it solid‐bound form, and structural rules that suggest ideal peptide design for acquiring objective peptide function.     

Combined screening of thymocytes using apoptosis-specific cDNA array and promoter analysis yields novel gene targets mediating TCDD-induced toxicity.

We have used pathway-specific cDNA arrays coupled with analysis of gene promoter regions to identify novel genes that may mediate the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the thymus. C57BL/6 mice were injected ip with 50 microg/kg TCDD, and 3, 6, or 24 h later, RNA was extracted from the thymus and subjected to microarray analysis. Several members of the TNF and TNFR family were induced following TCDD exposure, including receptor/ligand pairs Ltbeta-R/LIGHT, OX40/OX40L and TNF-alpha/TNFR1. In addition, Fas and CD30 were also upregulated. Pro-apoptotic bcl-2 gene family members Bax and Hrk, among others, were also induced, as were pro-survival bcl-2 family genes Bcl-x and Bcl-w. Cell-cycle regulator p21Cip1 was also induced. In addition, we analyzed the promoter regions of genes induced by TCDD for the presence of dioxin-responsive elements (DREs). The Fas and LIGHT gene promoters were found to contain DREs as analyzed by Matinspector Web-based search algorithm. Furthermore, binding of the aryl hydrocarbon receptor (AhR) to the DREs present on these genes was confirmed by chromatin immunoprecipitation. Given that several of the genes, including Fas, LIGHT, and CD30 are involved in negative selection of T cells in the thymus, our studies suggest that TCDD-induced upregulation of these genes may enhance negative selection leading to thymic atrophy.     

川芎嗪对儿茶酚胺诱导大鼠心肌损伤时Bcl-2和Fas蛋白表达的影响

目的 :观察川芎嗪对儿茶酚胺诱导大鼠心肌损伤时心肌细胞Bcl 2和Fas蛋白表达的影响。方法 :采用皮下多点注射异丙肾上腺素 (ISO ,5mg·kg- 1)每天 1次 ,连续 3d ,诱导大鼠心肌损伤 ,采用免疫组化方法检测Bcl 2和Fas蛋白的水平 ,并利用图像分析系统分析蛋白阳性表达区域平均光密度值。结果 :损伤组和川芎嗪保护组心肌细胞Bcl 2蛋白表达较对照组均显著升高 (P <0 .0 1 ) ,损伤组Fas蛋白水平较对照组也显著升高 (P <0 .0 1 ) ,川芎嗪保护组的Fas蛋白水平较损伤组显著下降 (P <0 .0 1 ) ,且Bcl 2 Fas比值也较损伤组和对照组明显增加。川芎嗪保护组的病理损伤程度明显低于损伤组。结论 :川芎嗪可抑制儿茶酚胺诱导的心肌损伤时心肌细胞中促凋亡基因Fas的表达 ,提高心肌细胞中Bcl 2 Fas比值     

Beliefs about fetal alcohol spectrum disorder among men and women at alcohol serving establishments in South Africa

Background: South Africa has one of the highest rates of fetal alcohol spectrum disorder (FASD) in the world. However, little is known about what men and women who attend alcohol serving establishments believe about alcohol use during pregnancy and how these beliefs may be related to alcohol use. Objectives: To understand FASD beliefs and related behaviors among men and women attending alcohol-serving establishments. Methods: We surveyed 1047 men (n?=?565) and women (n?=?482) -including pregnant women and men with pregnant partners- attending alcohol serving establishments in a township located in Cape Town, South Africa. Results: Among both pregnant (n?=?53) and non-pregnant (n?=?429) women, 54% reported drinking alcohol at least 2–4 times per month, and 57% reported having at least 3–4 alcohol drinks during a typical drinking session. Pregnant women were less likely to believe that they should not drink alcohol and that alcohol can harm a fetus when compared to non-pregnant women. Similar findings were observed between men with pregnant partners compared to men without pregnant partners. Among women, beliefs about how much alcohol pregnant women can safely drink were associated with self-reported alcohol use. Conclusions: Efforts to address FASD need to focus on understanding how men and women perceive alcohol use during pregnancy and situational factors that contribute to alcohol consumption among pregnant women attending alcohol serving establishments. Structural and individual-level interventions targeting women at alcohol serving establishments should be prioritized to mitigate alcohol use during pregnancy.     

铜绿假单胞菌上清液诱导J774细胞凋亡过程中Fas蛋白的表达

<正>铜绿假单胞菌(pseudomonas aeruginosa,PA)是医院内感染最常见的致病菌,一旦感染,治疗困难。巨噬细胞是机体重要的免疫防御细胞,细菌侵袭机体组织,首先要避免被巨噬细胞等机体免疫细胞的吞噬、清除。因此激发巨噬细胞凋亡对细菌而言很重要。近来研究显示,PA以剂量依赖和时间依赖的方式诱导U937巨噬细胞凋亡,且与线粒体途径有关[1]。本研究采用J774细胞作为巨噬细胞的模型[2],通过观察PA上清液诱导J774细胞凋亡过程中Fas蛋白的表达,进一步探讨PA上清液诱导J774细胞凋亡的致病机制,为开发新型抗感染药物提供重要的理论依据。