Objective: To determine the effects of hawthorn extract on serum lipid levels, pathological changes in aortic atherosclerosis plaque, inflammatory factors, and apoptosis-related protein and mRNA expression in apolipop...Objective: To determine the effects of hawthorn extract on serum lipid levels, pathological changes in aortic atherosclerosis plaque, inflammatory factors, and apoptosis-related protein and mRNA expression in apolipoprotein E gene knockout(ApoE^(-/-)) mice. Methods: Thirty-six ApoE^(-/-) mice were fed with a high-fat diet starting at the age of 8 weeks. Mice were randomly divided into 3 groups by a random number table including model group, hawthorn extract group, and simvastatin group, 12 mice in each group. Twelve 8-week-old C57BL/6 mice were fed a basic diet and served as control. The mice in the control and model groups were administered 0.2 mL saline daily, the mice in the hawthorn extract and simvastatin groups were administered with 50 mg/kg hawthorn extract or 5 mg/kg simvastatin daily for 16 weeks. After 16 weeks, plasma lipids including total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) were determined by an enzymatic assay. Aortic atherosclerotic lesions were observed by light microscopy, scanning and transmission electron microscopy, respectively. Plasma levels of monocyte chemoattractant protein-1(MCP-1), interleukin-1β(IL-1β), adiponectin(APN), and hypersensitive C-reactive protein(hs-CRP) were measured by enzyme-linked immunosorbent assay(ELISA). Protein and mRNA expressions of Bax and Bcl-2 in the aorta were assessed by Western blotting and quantitative real-time polymerase chain reaction(qR T-PCR), respectively. Results: Compared to the control group, the plasma levels of TC, TG and LDL-C were significantly increased and HDL-C were significantly decreased in the model group(P<0.01). Compared to the model group, treatment with hawthorn extract significantly decreased the plasma levels of TC, TG, and LDL-C and increased the plasma level of HDL-C in ApoE^(-/-)mice(P<0.01). The levels of MCP-1, IL-1β, and hs-CRP in the model group were significantly increased and APN was significantly decreased compared with the contr展开更多
目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling...目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling）法检测CIK细胞的凋亡.通过免疫细胞化学染色法检测ZK-75-1细胞中p53、p16、C-myc、Bcl-2及Bax的表达率.结果：HE染色显示,CIK细胞向ZK-75-1细胞靠近,形成典型的玫瑰花环状;肿瘤细胞的胞浆中出现颗粒状物,有的肿瘤细胞只见颗粒状碎片;而作为对照的乳腺癌细胞生长良好.TUNEL法检测显示,对照组细胞未染色或染呈均匀的淡蓝色;实验组凋亡的细胞缩小,核或核周染呈深蓝色.CIK细胞作用4～12 h ZK-75-1细胞的凋亡率上升,作用12～24h细胞的凋亡率下降,与对照组相比较有统计学意义（P＜0.01）.免疫细胞化学染色的结果表明,CIK细胞实验组p53、pi6、C-myc及Bcl-2蛋白随作用时间的延长均下降,Bax蛋白的表达上调,与对照组相比较均有统计学意义（P＜0.01）.结论：CIK细胞对乳腺癌细胞ZK-75-1杀伤作用的机制之一,可能与p53、p16、C-myc、Bcl-2蛋白表达的下调及Bax蛋白表达的上调有关,并与CIK细胞作用的时间关系密切.展开更多
目的:探讨姜黄素预处理的外泌体对肝癌细胞增殖、凋亡及细胞周期的影响,以期发现姜黄素治疗肝癌的新靶点及新途径。方法:采用CCK-8法检测不同浓度的姜黄素对肝癌细胞HUH-7活力的影响,并计算半数抑制浓度(IC)值。HUH-7细胞在姜黄素中暴露...目的:探讨姜黄素预处理的外泌体对肝癌细胞增殖、凋亡及细胞周期的影响,以期发现姜黄素治疗肝癌的新靶点及新途径。方法:采用CCK-8法检测不同浓度的姜黄素对肝癌细胞HUH-7活力的影响,并计算半数抑制浓度(IC)值。HUH-7细胞在姜黄素中暴露48 h,采取改良超高速离心法提取姜黄素外泌体,应用透射电镜、纳米颗粒追踪分析仪、蛋白印迹方法分别进行外泌体形态、粒径、浓度及标志蛋白的鉴定。采用PKH-26标记姜黄素外泌体,采用长时间动态细胞成像系统观察姜黄素外泌体被HepG2细胞摄取情况。将姜黄素外泌体与HepG2细胞共培养,采用CCK-8法测定细胞活力,流式细胞仪计算细胞周期,TUNEL法观察细胞凋亡的形态特点,WB法检测B细胞淋巴瘤因子-2(B cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白质(Bcl-2-related X protein,BAX)表达,判断细胞凋亡的情况。结果:姜黄素对HUH-7细胞48 h IC值为41.46μmol/L;经证实提取物为外泌体,姜黄素预处理能增加细胞外泌体的分泌,但对外泌体形态、表面标志物及粒径大小无影响;PKH-26外泌体示踪结果显示HepG2细胞能摄取姜黄素外泌体,姜黄素外泌体能明显抑制细胞活力(P<0.05),能将HepG2细胞阻滞于G/M和S期(P<0.05),使BAX蛋白表达上调,Bcl-2蛋白表达下调(P<0.01),能明显促HepG2细胞凋亡。结论:姜黄素预处理的外泌体能抑制肝癌细胞活力、阻滞细胞周期、促进细胞凋亡,表现出一定抗细胞增殖效应。展开更多
基金Supported by the National Natural Science Foundation of China(No.81573821)General Financial Grant from China Postdoctoral Science Foundation(No.2016M600176)Special Financial Grant from China Postdoctoral Science Foundation(No.2017T100147)
文摘Objective: To determine the effects of hawthorn extract on serum lipid levels, pathological changes in aortic atherosclerosis plaque, inflammatory factors, and apoptosis-related protein and mRNA expression in apolipoprotein E gene knockout(ApoE^(-/-)) mice. Methods: Thirty-six ApoE^(-/-) mice were fed with a high-fat diet starting at the age of 8 weeks. Mice were randomly divided into 3 groups by a random number table including model group, hawthorn extract group, and simvastatin group, 12 mice in each group. Twelve 8-week-old C57BL/6 mice were fed a basic diet and served as control. The mice in the control and model groups were administered 0.2 mL saline daily, the mice in the hawthorn extract and simvastatin groups were administered with 50 mg/kg hawthorn extract or 5 mg/kg simvastatin daily for 16 weeks. After 16 weeks, plasma lipids including total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) were determined by an enzymatic assay. Aortic atherosclerotic lesions were observed by light microscopy, scanning and transmission electron microscopy, respectively. Plasma levels of monocyte chemoattractant protein-1(MCP-1), interleukin-1β(IL-1β), adiponectin(APN), and hypersensitive C-reactive protein(hs-CRP) were measured by enzyme-linked immunosorbent assay(ELISA). Protein and mRNA expressions of Bax and Bcl-2 in the aorta were assessed by Western blotting and quantitative real-time polymerase chain reaction(qR T-PCR), respectively. Results: Compared to the control group, the plasma levels of TC, TG and LDL-C were significantly increased and HDL-C were significantly decreased in the model group(P<0.01). Compared to the model group, treatment with hawthorn extract significantly decreased the plasma levels of TC, TG, and LDL-C and increased the plasma level of HDL-C in ApoE^(-/-)mice(P<0.01). The levels of MCP-1, IL-1β, and hs-CRP in the model group were significantly increased and APN was significantly decreased compared with the contr
文摘目的：研究多种细胞因子诱导的杀伤细胞（cytokine-induced killing cells,CIK）对乳腺癌细胞株ZK-75-1的杀伤作用,并探讨其作用机制.方法：通过HE染色观察凋亡细胞ZK-75-1的形态学改变.应用TUNEL（TdT-mediated dUTPnick end labeling）法检测CIK细胞的凋亡.通过免疫细胞化学染色法检测ZK-75-1细胞中p53、p16、C-myc、Bcl-2及Bax的表达率.结果：HE染色显示,CIK细胞向ZK-75-1细胞靠近,形成典型的玫瑰花环状;肿瘤细胞的胞浆中出现颗粒状物,有的肿瘤细胞只见颗粒状碎片;而作为对照的乳腺癌细胞生长良好.TUNEL法检测显示,对照组细胞未染色或染呈均匀的淡蓝色;实验组凋亡的细胞缩小,核或核周染呈深蓝色.CIK细胞作用4～12 h ZK-75-1细胞的凋亡率上升,作用12～24h细胞的凋亡率下降,与对照组相比较有统计学意义（P＜0.01）.免疫细胞化学染色的结果表明,CIK细胞实验组p53、pi6、C-myc及Bcl-2蛋白随作用时间的延长均下降,Bax蛋白的表达上调,与对照组相比较均有统计学意义（P＜0.01）.结论：CIK细胞对乳腺癌细胞ZK-75-1杀伤作用的机制之一,可能与p53、p16、C-myc、Bcl-2蛋白表达的下调及Bax蛋白表达的上调有关,并与CIK细胞作用的时间关系密切.
文摘目的:探讨姜黄素预处理的外泌体对肝癌细胞增殖、凋亡及细胞周期的影响,以期发现姜黄素治疗肝癌的新靶点及新途径。方法:采用CCK-8法检测不同浓度的姜黄素对肝癌细胞HUH-7活力的影响,并计算半数抑制浓度(IC)值。HUH-7细胞在姜黄素中暴露48 h,采取改良超高速离心法提取姜黄素外泌体,应用透射电镜、纳米颗粒追踪分析仪、蛋白印迹方法分别进行外泌体形态、粒径、浓度及标志蛋白的鉴定。采用PKH-26标记姜黄素外泌体,采用长时间动态细胞成像系统观察姜黄素外泌体被HepG2细胞摄取情况。将姜黄素外泌体与HepG2细胞共培养,采用CCK-8法测定细胞活力,流式细胞仪计算细胞周期,TUNEL法观察细胞凋亡的形态特点,WB法检测B细胞淋巴瘤因子-2(B cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白质(Bcl-2-related X protein,BAX)表达,判断细胞凋亡的情况。结果:姜黄素对HUH-7细胞48 h IC值为41.46μmol/L;经证实提取物为外泌体,姜黄素预处理能增加细胞外泌体的分泌,但对外泌体形态、表面标志物及粒径大小无影响;PKH-26外泌体示踪结果显示HepG2细胞能摄取姜黄素外泌体,姜黄素外泌体能明显抑制细胞活力(P<0.05),能将HepG2细胞阻滞于G/M和S期(P<0.05),使BAX蛋白表达上调,Bcl-2蛋白表达下调(P<0.01),能明显促HepG2细胞凋亡。结论:姜黄素预处理的外泌体能抑制肝癌细胞活力、阻滞细胞周期、促进细胞凋亡,表现出一定抗细胞增殖效应。
