组蛋白脱乙酰酶2介导的组蛋白乙酰化失衡在压力过载性心肌重构中的作用

目的：探讨组蛋白脱乙酰酶2(histone deacetylase 2,HDAC2)介导的组蛋白乙酰化修饰失衡在压力过载性心肌重构中的调控作用。方法：选取SPF级成年昆明小鼠,通过部分结扎胸主动脉（transverse aortic constriction,TAC）建立小鼠压力过载性心肌重构模型,应用表没食子儿茶素没食子酸酯（epigallocatechin gallate,EGCG）腹腔注射干预模型小鼠。根据随机数字表法将小鼠分为正常组、TAC组、溶剂组、假手术组和EGCG+TAC组,每组6只小鼠。收集各组小鼠心脏组织进行如下检测：Western blot检测HDAC2、第27位赖氨酸乙酰化的组蛋白H3(acetylated histone H3 at lysine 27,H3K27ac)、心肌细胞增强因子2A(myocyte enhancer factor 2A,MEF2A)和脑钠肽（brain natriuretic peptide,BNP）蛋白表达水平;超声心动图检测小鼠心功能;ES420生物系统检测小鼠左室收缩末期压力（left ventricular end-systol...     

EGCG抑制肝癌细胞株HepG2增殖及HIF-1α/VEGF的表达

目的:研究表没食子儿茶素没食子酸酯(EGCG)抑制肝癌生长的分子机制。方法:采用MTT方法观察EGCG对HepG2细胞株增殖的影响;利用体外厌氧培养技术,通过不同剂量的EGCG进行干预,检测肝癌细胞HIF-1α/VEGF的基因转录和蛋白表达;将肝癌细胞种植于裸鼠体内,观察EGCG对肿瘤生长的抑制作用,并且测定HIF-1α/VEGF的表达。结果:体外试验显示EGCG对肝癌细胞株HepG2的增殖有明显抑制作用;明显下调HIF-1α/VEGF蛋白的表达,而对HIF-1α基因转录影响不大;裸鼠体内试验中观察到EGCG对肿瘤生长有明显抑制作用,抑制率达39.8%±5.1%。结论:EGCG可明显抑制肝癌细胞的增殖,干预HIF-1α/VEGF信号转导通路可能是其抑制肿瘤生长的主要机制之一。     

链状酰胺类化合物B07的体外杀精效果及其对精子质膜的影响

目的:研究一种链状酰胺类化合物(B07)的体外杀精作用及其对精子质膜的影响。方法:上游收集高活力人类精子分别与0μg/ml(对照组),5~640μg/ml B07作用30 min,随后用计算机辅助精液分析仪分析记录精子的活率及运动参数;用160μg/ml B07与精子作用,记录5 min、30 min、60 min和120 min时精子活动率;处理后的精子一部分经SYBR-14/PI双染后用荧光显微镜观察,另一部分通过电子显微镜来观察质膜的变化情况;并以壬苯醇醚(N9)作为阳性对照。结果:随着B07浓度的升高,精子活率和活力呈下降趋势,最低有效浓度(MEC)为640μg/ml;荧光显微镜下经B07处理的精子呈橙色,而N9处理的精子呈红色;电子显微镜显示B07组精子头部肿胀起泡,尾部针孔样损伤,N9组精子呈溶解性改变。结论:B07具有体外杀精作用,且该作用有时间和浓度依赖性,且其对精子质膜的影响小于N9。     

圆锥菝葜的化学成分研究

目的研究圆锥菝葜（Smilax bracteata）的化学成分。方法利用60%乙醇回流提取,大孔吸附树脂和硅胶柱色谱等方法进行分离纯化,根据化合物的光谱数据鉴定结构。结果从圆锥菝葜中分离得到5个化合物,分别鉴定为光叶菝葜苷（Ⅰ）、白藜芦醇（Ⅱ）、落新妇苷（Ⅲ）、新异落新妇苷（Ⅳ）、儿茶素（Ⅴ）。结论化合物Ⅰ~Ⅴ为首次从该种植物分离得到。     

苯丙氨酸二肽类化合物Y101对正常小鼠的免疫调节作用

目的 探讨苯丙氨酸二肽类化合物Y101对正常小鼠的免疫调节作用.方法 利用免疫学相关技术对Balb/C小鼠对照组、苯丙氨酸二肽类化合物Y101低、高(25、100mg/kg)剂量组的碳粒廓清指标进行测定,并对昆明小鼠Y101低、中、高(25、50、100mg/kg)剂量组的血清溶血素水平及脏器指数进行测定.结果 与空白...     

Protective effects of （-）-epigallocatechin-3-gallate on D-galactose-induced neuronal apoptosis in mice

BACKGROUND： The neuroprotective effects of （-）-epigallocatechin-3-gallate （EGCG）, the main polyphenolic constituent of green tea, have been widely reported. However, the action mechanisms, in particular in D-galactose-induced aging mice, remain poorly understood. OBJECTIVE： The present study investigated the protective effects of EGCG on D-galactose-induced hippocampus neuronal apoptosis in aging mice, as well as the relationship with expression of p751CD, JNK2, and p53 proteins. DESIGN, TIME AND SETTING： A randomized, controlled, molecular biological, animal experiment was performed at the Laboratory of Pharmacology, Pharmaceutical College of China Medical University, China, from September 2006 to July 2008. MATERIALS： D-galactose and EGCG （Sigma, USA）, as well as terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling （TUNEL） In Situ Cell Apoptosis Detection Kit （Promega, USA）, were used in this study. METHODS： A total of 64 mice were equally and randomly divided into D-galactose model, low-dose EGCG, high-dose EGCG, and control groups. Mice in the D-galactose model, low-dose EGCG, and high-dose EGCG groups were subcutaneously injected with 3% D-galactose （150 mg/kg）, daily for 6 weeks, to establish a mouse model of aging. Mice in the control group were treated with saline （5 mL/kg）. At 3 weeks following injection, mice in the low-dose EGCG and high-dose EGCG groups were orally administered EGCG at a dose of 2 mg/kg and 6 mg/kg, respectively, once a day, for 4 consecutive days. Mice in the control and D-galactose model groups received distilled water （5 mL/kg）. MAIN OUTCOME MEASURES： Memory function was evaluated using a step-through passive avoidance test. Neuronal apoptosis in the mouse hippocampus was detected using TUNEL staining. Expression levels of the intracellular domain of the p75 neurotrophin receptor （p75NTR）-p751CD, JNK2, and p53 proteins in the hippocampus were determined using Western blot analysis. RESULTS： The aging mouse model was induced by subcutaneous injection of D-galactose, which resulted in obvious memory impairment, increased apoptotic index, and increased protein expression levels of p751CD, JNK2, and p53 in the hippocampus, compared with control mice （P 〈 0.01）. Oral EGCG administration （2 or 6 mg/kg） for 4 weeks significantly improved levels of memory deficit in the aging mice and reduced apoptotic indices and protein expression levels of p751CD, JNK2, and p53 in the mouse hippocampus （P 〈 0.01）. CONCLUSION： Results from this study demonstrated increased protein expression levels of p751CD, JNK2, and p53, as well as increased hippocampal neuronal apoptosis in a D-galactose-induced mouse model of aging. EGCG provided protective effects against D-galactose-induced neuronal apoptosis in the hippocampus by reducing protein expression levels of p751CD, JNK2, and p53 proteins in the hippocampus of aging mice.     

白诺菌素类化合物的结构鉴定及核磁共振归属的研究

环二肽类物质在生物界中广泛存在.生物学研究表明,环二肽类物质有广泛的生物学活性[1],其中化合物cyclo(Δisoprenyl His-Δphe)在抗肿瘤方面有较强的作用,比现在临床用药(如紫杉醇)有更强的抗肿瘤活性[2].     

HPCE法测定蛹虫草及其培养物中核苷类化合物的含量

目的　建立蛹虫草中核苷类化合物的高效毛细管电泳测定法 ,比较天然蛹虫草及其培养物中核苷类化合物的含量。方法　在HV - 30 1高效毛细管电泳仪上进行定量分析 ,采用石英毛细管柱 ,以pH9.0的 4 0mmol·L-1 硼砂溶液作为电极缓冲液 ;分离电压为 1 6KV ;检测波长为 2 5 4nm。结果　天然蛹虫草中核苷类化合物主要集中在子座部分。人工培养蛹虫草子座和菌丝中虫草素的含量分别是天然蛹虫草子座中的 5 .0 4倍和 1 0 .85倍 ;液体培养蛹虫草菌丝中腺嘌呤、腺苷、尿苷、鸟苷含量均高于固体培养蛹虫草子座。结论　天然蛹虫草及其培养物中核苷类化合物的种类和含量具有较大的差异。高效毛细管电泳法可以用于蛹虫草的定量分析 ,具有分离度高、简便、准确 ,样品消耗少等特点     

生肌散质控方法的研究

目的 建立生肌散的质量标准。方法 采用TLC法对处方中血竭、冰片进行鉴别,用HPLC法测定方中君药儿茶中儿茶素的含量。结果 在TLC色谱中,能检出血竭、冰片。儿茶素在0．1762～3．524μg范围内,浓度与峰面积线性关系良好（r=0．9994）,样品的平均加样回收率为103．06％,RSD为0．39％（n=6）。结论 本文建立的方法对方中3味药材能准确、快速地进行定性、定量检测,可用于生肌散的质量控制。