Changes of NF—kB,p53,Bcl—2 and caspase in apoptosis induced by JTE—522 in human gastric adenocarcinoma cell line AGS cells：role of reactive oxygen species

AIM:To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process,and to investigate the changes in NF-kB,p53,bcl-2 and caspase in the apoptosis process.METHODS:Cell culture,MTT,Electromicroscopy,agarose gel electrophoresis,lucigenin,western blot and electrophoretic mobility shift assay(EMSA)analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms.RESULTS:JTE-522 inhitied the growth of AGS cells and induced the apoptosis,Lucigenin assay showed thd generation of ROS in cells under incubation with JTE-522,The increased ROS generation might Contribute to the induction of AGS cells to apoptosis.EMSA and Western blot revealed that NF-kB activity was almost completely inhihbited by preventing the degradation of IKBα,Additionally,by using Western blot we confirmed that the level of bcl-2 was decreased,whtereas p53 showed a great inmcrease following JTE-522 treatment.Their changes were in a dose-dependent manner. CONCLUSION：These findings suggest that reactive oxygen species,NF-kB,p53,bcl-2 and caspase-3 may play an important role in the induction of apoptosis,in ags cells after treatment with JTE-522.     

肝素和阿霉素对人鼻咽癌CNE2细胞增殖与凋亡的协同作用

目的:观察肝素联合阿霉素对人鼻咽癌细胞的=增殖与凋亡的影响及其可能的分子机制.方法:用MTT法、流式细胞术观察肝素联合阿霉素对人鼻咽癌细胞增殖及细胞周期的影响;而用TUNEL、琼脂糖凝胶电泳、Westem blot等方法观察肝素与阿霉素联合应用对人鼻咽癌细胞凋亡的作用.结果:肝素与阿霉素联合应用后对鼻咽癌细胞的生长抑制及其凋亡具有显著增强作用,同阿霉素单独应用于诱导细胞凋亡相比,低剂量的肝素与阿霉素联合应用后发现:TUNEL阳性细胞从12.6％±1.1％增加到65.7％±1.3％;G_0/G_1期细胞阻止,S期细胞明显减少,DNA“梯形”变化更加明显;Bax/Bcl-2的比率及p~(53)和p~(21)基因的表达明显升高.结论:肝素与阿霉素对人鼻咽癌细胞在抗增殖及促凋亡方面有协同作用,这可能与Bax/Bcx-2比率的升高及p~(53)和p~(21)的表达上调有关.     

Inhibition of CYP3A-mediated oxidation in human hepatic microsomes by the dietary derived complex phenol, gallic acid

Plant polyphenols, such as gallic acid, have been reported to have a range of biological activities including antimutagenic effects. Previously, we reported that gallic acid (3,4,5-trihydroxybenzoic acid), an agent found in wine and tea, inhibits androstenedione 6beta-hydroxylase activity (Ki 70 microm), a cytochrome P450 (CYP3A) marker in human liver microsomes. The pre-incubation of gallic acid (100 microM) with human liver microsomes in the absence of NADPH, as compared with the presence of NADPH, before assay of androstenedione 6beta-hydroxylase activity significantly increased the inhibitory effects of the gallic acid (0.03 +/- 0.03 nmol (mg microsomal protein)(-1) min(-1) compared with 0.20 +/- 0.06 nmol (mg microsomal protein) (-1) min(-1) (P < 0.05)). The antioxidant ascorbic acid and the radical scavenger glutathione prevented this observed increase in inhibition. Removal of gallic acid-derived products from the incubation completely restored CYP3A activity. In contrast, the activities of CYP1A and CYP2E, and non-CYP mediated reductive microsomal 17beta-hydroxysteroid dehydrogenase activity were refractory to inhibition by gallic acid.     

Noggin反义寡核苷酸对大鼠空间学习与记忆的影响

目的：研究noggin反义寡核苷酸对大鼠空间学习与记忆的影响．方法：用原位杂交技术检测大鼠海马内noggin mRNA的表达,Morris水迷宫检测大鼠空间学习与记忆．结果：Morris水迷宫训练引起大鼠海马齿回(DG)与CA3区noggin mRNA阳性神经元数明显增多．侧脑室注射noggin反义寡核苷酸,不仅抑制Morris水迷宫训练引起的海马noggin mRNA阳性神经元数增多,同时阻滞学习与记忆的形成．正义noggin寡核苷酸无此效应．结论：Noggin作为一胚胎基因,表达于成年大鼠的海马,并与其空间学习记忆行为有关．     

尼美舒利抑制小鼠荷瘤肝癌的生长:提高Bax/Bc 1-2表达的比值(英文)

目的:观察尼美舒利对小鼠荷瘤抑制和凋亡诱导作用并探讨其可能的分子机制。方法:雄性小鼠给予尼美舒利10mg/kg,20mg/kg和40mg/kg各灌胃21天,用电镜,流式细胞术,琼脂糖电泳观察尼美舒利诱导肿瘤细胞凋亡的作用,放射免疫法测定瘤组织PGE_2含量,Western blot观察COX-2,c-myc,Bax和Bcl-2的表达.结果:尼美舒利能抑制小鼠荷瘤生长,且具有剂量依赖性(14％到62％),高剂量时,尼美舒利诱导的细胞凋亡率是(51.3±1.5)％,DNA“梯形”变化更加明显,PGE_2含量明显降低,Westernblot显示COX-2和Bcl-2表达降低,Bax表达增加,c-myc无变化。结论:尼美舒利对荷瘤肝癌的抑制作用可能与抑制COX-2而降低PGE_2含量和上调Bax/Bcl-2比值诱导瘤细胞凋亡有关。     

JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

AIM: To investigate the role of the mitochondrial pathway inJTE-522-induced apoptosis and to investigate therelationship between cytuchrome C release, caspaee activityand loss of mitochondrial membrane potential (△ψm).METHODS: Call culture, cell counting, ELISA assay,TUNEL, flow cymetry, Western blot and fluorometric assaywere employed to investigate the effect of JTE-522 on cellproliferation and apoptosis in AGS cells and relatedmolecular mechanism. RESULTS: JrE-522 inhibited the growth of AGS cells andinduced the apoptosis. Caspases 8 and 9 were activatedduring apoptosis as judged by the appearance of cleavageproducts from procaspase and the caspase activities tocleave specific fluorogenic substrates. To elucidate whetherthe activation of caspases 8 and 9 was required for theapoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed thatcaspase inhibitors significantly inhibited the apoptosisinduced by JTE-522. In addition, the membranetranslocation of Bax and cytosolic release of cytochrome Caccompanying with the decrease of the uptake of Rhodamin123, were detected at an early stage of apoptosis.Furthermore, Bax translocation, cytochrome C release, andcaspase 9 activation were blocked by Z-VAD. fmk and Z-IETD-CHO.CONCLUSION: The present data indicate a crucialassociation between activation of caspases 8, 9,cytochrome C release, membrane translocation of Bax, lossof△ψm and JTE-522-induced apoptosis in AGS cells.     

Studies on the effects of polydextrose intake on physiologic functions in Chinese people

BACKGROUND: Previous studies showed that polydextrose has physiologic effects similar to those of dietary fiber. OBJECTIVE: Ingestion of 4, 8, and 12 g polydextrose/d was studied to determine the physiologic effects in Chinese subjects. DESIGN: In a placebo-controlled, randomized, double-blind study, we evaluated the effects of polydextrose ingestion on clinical biochemistry indexes, glycated hemoglobin, glucose tolerance, the glycemic index, bowel function, stool weight and pH, short-chain fatty acid production, fecal microflora, and cecal mucosa cell proliferation. RESULTS: Polydextrose had no significant effect on blood biochemistry indexes. Ingestion of 12 g polydextrose plus 50 g glucose resulted in a glycemic index of 89% (compared with a glycemic index of 100% after ingestion of 50 g glucose). Bowel function (frequency and ease of defecation) improved significantly and there were no reports of abdominal distention, abdominal cramps, diarrhea, or hypoglycemia. Fecal weight (wet and dry) increased and fecal pH decreased proportionally to polydextrose intake. Short-chain fatty acid production-notably that of butyrate, isobutyrate, and acetate-increased with polydextrose ingestion. There were substantial changes in fecal anaerobes after polydextrose intake. BACTEROIDES: species (B. fragilis, B. vulgatus, and B. intermedius) decreased, whereas LACTOBACILLUS: and BIFIDOBACTERIUM: species increased. The cecal mucosa whole-crypt labeling index increased, with colonocyte proliferation mainly occurring in base compartments, which provided an indirect confirmation of butyrate production in the colon. CONCLUSION: Polydextrose ingestion had significant dietary fiber-like effects with no laxative problems.     

HPLC法测定阿霉素毫微粒经肝动脉灌注后大鼠体内的早期动态分布

目的　检测阿霉素毫微粒 (NADM)经肝动脉灌注后 ,在大鼠体内的药物分布 ,并与游离阿霉素 (FADM)相比较 .方法　以 α-聚氰基丙烯酸正丁酯为载体材料 ,制备NADM;SD纯系大鼠 2 4只 ,随机分为两组 ,每组 1 2只动物 ,经肝动脉分别注入 NADM与 FADM,药物剂量 2 mg·kg- 1 ,药物浓度 1 .0 g· L- 1 .每组于给药后 1 ,5 ,1 5 h各处死 4只大鼠 ,分别提取血浆、肝、心、脾、肺、肾样品 ,以反相高效液相色谱荧光检测法 (RP- HPL C)测定药物浓度 .结果　与FADM组相比 ,1 ,5 ,1 5 h时 NADM组大鼠肝、脾中阿霉素浓度显著增高 ,而血浆、肺、肾中阿霉素浓度降低 .心脏药物浓度给药后 1 h,5 h FADM组高于 NADM组 ,1 5 h时低于NADM组 .结论　 NADM肝动脉给药后改变了阿霉素的体内分布特征 ,对肝、脾表现出明显的靶向性 ,而血、心、肺、肾中的药物分布减少 .     

重力液流系统和主动控制液流系统对白内障超声乳化术中累积释放能量及术后视力恢复和角膜内皮参数的影响

目的　对比重力液流系统和主动控制液流系统对白内障超声乳化手术中累积释放能量（cumulative dissipated energy,CDE）及术后视力恢复、角膜内皮参数的影响。方法　选取2017年10月至12月于首都医科大学附属复兴医院眼科行白内障手术患者96例（96眼）。所有患者均患有年龄相关性白内障,LOCS III晶状体核透光性和晶状体核颜色评分为4.0~6.9分。依照液流系统选择情况分为:重力组（48例48眼）,使用Centurion重力液流系统;主控组（48例48眼）,使用Centurion主动控制液流系统。术中记录每例患者CDE;术后1周、1个月、3个月复诊,行眼科常规检查,包括视力、眼压、裂隙灯、眼底镜等;所有患者术前行非接触角膜内皮镜检查,记录角膜内皮细胞密度（endothelial cell density,ECD）、内皮细胞变异系数（coefficient of variance,CV）、角膜六角形内皮细胞比率（percentage of hexagonal endothelial cells,6A）及中央角膜厚度（central corneal thickness,CCT）。每次复查均行角膜内皮镜检查获取ECD、内皮细胞CV、6A、CCT等数据。结果　术前重力组和主控组患者的年龄、性别、晶状体核透光性、晶状体核颜色评分比较,差异均无统计学意义（均为P＞0.05）。术前两组患者最佳矫正视力比较,差异无统计学意义（P＞0.05）,术后1周主控组患者最佳矫正视力较重力组明显提高,差异有统计学意义（P＜0.05）,术后1个月和3个月两组间最佳矫正视力差异均无统计学意义（均为P＞0.05）。重力组与主控组术中CDE差异无统计学意义（P＞0.05）。总体CDE均与术前晶状体核分级呈正相关性（晶状体核透光性:r=0.703,P=0.001;晶状体核颜色评分:r=0.804,P=0.000）,术前晶状体核分级越高术中CDE越大。重力组和主控组术前ECD、内皮细胞CV、6A、CCT相似,差异均无统计学意义（均为 P＞0.05）。术后1周ECD、内皮细胞CV、6A比较,两组间差异均无统计学意义（均为P＞0.05）,但重力组CCT［（570.60±34.99）μm］明显大于主控组［（554.67±30.71）μm］,差异有统计学意义（P＜0.05）;术后1个月和3个月两组ECD、内皮细胞CV、6A、CCT比较,差异均无统计学意义（均为P＞0.05）。结论　Centurion超声乳化手术系统中重力液流系统及主动控制液流系统对术中CDE及角膜内皮的影响相似,主动控制液流系统能够更有效地减轻术后角膜水肿,使患者在术后早期视力恢复更迅速。     

SC58125抑制IκBα降解调节TNF-α诱导的HT29细胞凋亡

目的观察非甾体类抗炎药SC58125与TNFα是否具有协同诱导HT29细胞的凋亡作用,同时探讨其可能的分子机制。方法用MTT、琼脂糖凝胶电泳及流式细胞术,观察SC58125/TNFα对HT29细胞增殖与凋亡的影响;用EMSA及Westernblot检测转录因子NFκB的结合活性及IκBα的表达。结果SC58125与TNFα在抑制HT29细胞增殖及诱导其凋亡方面具有明显的协同作用,TNFα使HT29细胞生长受到明显抑制,DNA发生典型的“梯型”变化;SC58125可明显增强TNFα抑制细胞增殖和诱导细胞凋亡作用,使HT29细胞凋亡率从11.2%±1.1%增加到53.9%±2.1%。凋亡过程中伴随Caspase3活性的激活;经TNFα刺激后的HT29细胞,IκBα迅速降解,NFκB的结合活性大大提高;而加入SC58125后,IκBα降解及NFκB的结合活性均明显受到抑制。结论SC58125与TNFα在诱导HT29细胞的凋亡方面具有明显的协同作用,这可能与SC58125抑制IκBα降解和激活Caspase有关。