海风藤新木脂素类成分对缺血再灌注鼠脑损伤的保护作用

目的 探讨血小板活化因子 (PAF)受体拮抗剂海风藤新木脂素类成分对鼠脑梗死面积、局部脑血流量 (rCBF)及神经功能缺失征象的影响 ,并与传统的PAF受体拮抗剂银杏苦内酯相比较。方法 利用TTC染色结合球积仪、氢清除法组织血流计和临床神经功能评分标准等方法 ,分别对缺血、缺血再灌注及海风藤新木脂素类成分、银杏苦内酯治疗组鼠脑梗死面积、rCBF、神经功能缺失征象进行观察和比较。结果 海风藤新木脂素类成分、银杏苦内酯药物治疗组脑梗死面积较缺血再灌组明显减小 ,同时药物能改善缺血侧皮质rCBF的下降 ,并显著减轻脑缺血后的神经功能缺损。

海风藤中抗血小板活化因子成分2,5-二芳基四氢呋喃型木脂体的研究

从胡椒科植物中药海风藤茎中发现3种抗血小板活化因子成分,经HPLC分离和光谱分析鉴定为galgravin, galbelgin, veraguensin。

配伍应用海风藤、络石藤、忍冬藤治疗顽固性过敏性紫癜经验

过敏性紫癜是多种因素引起的系统性血管炎病变,主要累及皮肤、胃肠道、关节和肾脏等多器官。丁樱教授在过敏性紫癜的治疗上善用藤类药物,取其疏风通络、活血养血之功效,配伍使用海风藤、络石藤、忍冬藤三药,临床上取得了显著疗效。

Inhibition of beta-site amyloid precursor protein-cleaving enzyme and beta-amyloid precursor protein genes in SK-N-SH cells

BACKGROUND： Previous studies have demonstrated that Piper futokadsura stem selectively inhibits expression of amyloid precursor protein （APP） at the mRNA level. In addition, the piperlonguminine （A） and dihydropiperlonguminine （B） components （1 ： 0.8）, which can be separated from Futokadsura stem, selectively inhibit expression of the APP at mRNA and protein levels. OBJECTIVE： Based on previous findings, the present study investigated the effects of β-site amyloid precursor protein cleaving enzyme （BACE1） and APP genes on the production of β-amyloid peptide 42 （Aβ42） in human neuroblastoma cells （SK-N-SH cells） using small interfering RNAs （siRNAs） and A/B components separated from Futokadsura stem, respectively. DESIGN, TIME AND SETTING： A gene interference-based randomized, controlled, in vitro experiment was performed at the Key Laboratory of Cardiovascular Remodeling and Function Research, Ministries of Education and Public Health, and Institute of Pharmacologic Research, School of Pharmaceutical Science ＆ Department of Biochemistry, School of Medicine, Shandong University between July 2006 and December 2007. MATERIALS： SK-N-SH cells were provided by Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai, China; mouse anti-human BACE1 monoclonal antibody was purchased from R＆D Systems, USA; mouse anti-human APP monoclonal antibody was purchased from Cell Signaling Technology, USA; and horseradish peroxidase （HRP）-conjugated goat anti-mouse IgG was provided by Sigma, USA. METHODS： The human BACE1 cDNA sequence was obtained from NCBI website （www.ncbi.nlm.nih.gov/sites/entrez）. Three pairs of siRNAs, specific to human BACE1 gene, were synthesized through the use of Silencer pre-designed siRNA specification, and were transfected into SK-N-SH cells with siPORT NeoFX transfection agent to compare the effects of different concentrations of siRNAs （10-50 nmol/L） on SK-N-SH cells. Futokadsura stem was separated and purified with chemical methods, and the crystal was composed of A/B components, with an A to B ratio of 1：0.8. The A/B （1 ： 0.8） components were added to the SK-N-SH cells at different concentrations （13.13, 6.56, and 3.28 mg/mL）. MAIN OUTCOME MEASURES： Using RT-PCR and Western blot methods, BACE1 and APP expression at mRNA and protein levels was detected in SK-N-SH cells following treatment with different siRNAs and concentrations of Futokadsura stem-separated A/B components, respectively. Altered Aβ42 secretion by SK-N-SH cells was determined by ELISA. RESULTS： BACE1 mRNA and protein levels were significantly suppressed by 40 and 50 nmol/L siRNAs at 48 hours post-transfection. A/B components （1 ： 0.8）, which were separated from Futokadsura stem, selectively inhibited mRNA and protein expression of APP in SK-N-SH cells. Aβ42 secretion by SK-N-SH cells was significantly decreased following treatment with siRNAs or A/B components. CONCLUSION： Inhibition of BACE1 and APP genes by various materials and methods efficiently decreased production of Aβ42.

海风藤新木脂素成分对缺血鼠脑细胞间粘附分子-1及其mRNA表达的影响

目的 探讨海风藤新木脂素类成分对缺血鼠脑细胞间粘附分子 1(ICAM 1)及其mRNA表达的影响。方法 建立大鼠大脑中动脉闭塞 /再灌注模型 ,应用RT PCR及免疫组织化学的方法 ,观察海风藤新木脂素复合物以及银杏内脂对缺血鼠脑ICAM 1及其mRNA表达的影响。结果 免疫组织化学染色显示ICAM 1在假手术组鼠脑呈低表达 ;缺血 90min表达上调 ;缺血 90min再灌 2 4h后ICAM 1表达较前差异有非常显著性 (P <0 .0 1) ;而海风藤新木脂素类成分以及银杏内脂治疗组于相同时限鼠脑ICAM 1表达与单纯缺血、缺血再灌注组比较显著下调(P <0 .0 1)。假手术组鼠脑ICAM 1mRNA呈低表达 ;缺血 90min/再灌 2 4h其表达水平较对照组显著上调 (P <0 .0 1) ;药物治疗组于相同时限ICAM 1mRNA的表达显著下调 (P <0 .0 1)。结论 海风藤新木脂素类成分以及海风藤酮能明显降低缺血脑组织ICAM 1及其mRNA的表达 ,从而减轻脑缺血后炎性病理损害 。