杜仲带腋芽茎段组织培养的研究

目的研究不同培养条件下杜仲带腋芽茎段愈伤组织的诱导和生长情况,以及腋芽的生长情况。方法以带腋芽的杜仲茎段为外植体,设计9种不同的培养基配方,为了防止褐化,在23.4℃下培养箱中暗培养,30 d后调查出愈率及腋芽的生长情况。结果 NAA浓度从0.05~0.5 mg.L-1,随着浓度增加,腋芽生长受抑制;6-BA浓度从0.3~1.0 mg.L-1,随着浓度增加,愈伤组织块增大,达到1.0 mg.L-1时愈伤组织的生长受到抑制;本实验中最佳的杜仲愈伤组织诱导和生长培养基配方为6号培养基:B5+0.5 mg.L-1NAA+0.3 mg.L-16-BA。腋芽生长最佳培养基配方为4号培养基:B5+0.05 mg.L-1NAA+0.5 mg.L-16-BA。结论本试验的结果对杜仲的生物技术利用有一定的参考价值。     

Network pharmacology explores the mechanisms of Eucommia ulmoides cortex against postmenopausal osteoporosis

Postmenopausal osteoporosis (PMOP) has become one of most frequent chronic disease worldwide with aging population. Eucommia ulmoides cortex (EU), a traditional Chinese medicine, has long since been used to treat PMOP. The aim of this study is to explore pharmacological mechanisms of EU against PMOP through using network pharmacology approach.The active ingredients of EU were obtained from Traditional Chinese Medicine System Pharmacology database, and target fishing was performed on these ingredients in UniProt database for identification of their relative targets. Then, we screened the targets of PMOP using GeneCards database and DisGeNET database. The overlapping genes between PMOP and EU were obtained to performed protein–protein interaction, Gene Ontology analysis, Kyoto encyclopedia of genes, and genomes analysis.Twenty-eight active ingredients were identified in EU, and corresponded to 207 targets. Also, 292 targets were closely associated with PMOP, and 50 of them matched with the targets of EU were considered as therapeutically relevant. Gene ontology enrichment analysis suggested that EU exerted anti-PMOP effects via modulating multiple biological processes including cell proliferation, angiogenesis, and inflammatory response. Kyoto encyclopedia of genes and genomes enrichment analysis revealed several pathways, such as PI3K-AKT pathway, mitogen-activated protein kinase pathway, hypoxia-inducible factors-1 pathway, tumor necrosis factor pathway, and interleukin-17 pathway that might be involved in regulating the above biological processes.Through the method of network pharmacology, we systematically investigated the mechanisms of EU against PMOP. The multi-targets and multi-pathways identified here could provide new insights for further determination of more exact mechanisms of EU.     

Network pharmacology-based pharmacological mechanism prediction on Eucommia ulmoides against rheumatoid arthritis

Rheumatoid arthritis (RA) is a common chronic autoimmune disease characterized by synovial inflammation and progressive joint destruction. Eucommia ulmoides (EU) is a kidney-tonifying Chinese medicine that has been applied to treat RA for decides. The present study aims to explore pharmacological mechanisms of EU against RA using network pharmacology approach. Traditional Chinese Medicine Systems Pharmacology (TCMSP) database was used to screen active ingredients of EU, and their relative targets were fished from UniProt database. RA-related targets were screened from GeneCards database and DisGeNET database. The overlapping genes between EU and RA were identified by Venn diagram, and further analyzed for protein-protein interaction (PPI), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG). Fifty active ingredients were identified in EU, and corresponded to 207 targets. Meanwhile, 499 targets were closely associated with RA development. A total of 50 overlapping genes between EU and RA were identified, which were regarded as therapeutically relevant. GO enrichment analysis indicated that EU exerted antiRA effects depending on regulating multiple biological processes including inflammatory response, oxidative stress, cell apoptosis and matrix catabolism. Several key pathways such as TNF pathway, IL-17 pathway, T cell receptor pathway, NOD-like receptor pathway and Toll-like receptor pathway, were involved in the above biological processes. Network pharmacology revealed that EU exerts therapeutic effects on RA through multi-ingredients, multi-targets and multi-pathways, which provides basis for its clinical application and promising directions for subsequent research.     

“杜仲-当归”介导Wnt/β-catenin信号转导调控MMP-13表达防治关节炎的作用机制研究

目的 研究“杜仲-当归”介导Wnt/β-catenin信号转导调控MMP13表达防治骨性关节炎（osteoarthritis,OA）的作用机制。方法 取SD大鼠,随机分为Sham组、OA组、OA+补肾组（杜仲40 mg·kg^-1）、OA+活血组（当归40 mg·kg^-1）、OA+补肾活血组（杜仲-当归药液40 mg·kg^-1）、OA+阳性药物组（西乐葆10 mg·kg^-1+奥泰灵1.2 mg·kg^-1）共6组。OA模型制备成功后,各组大鼠给予对应药物治疗,连续给药8周。番红O染色检测软骨组织变化,免疫组织化学检测软骨组织中MMP-13表达,ELISA检测大鼠血清IL-6、IL-8含量,Western blotting检测软骨组织中MMP-3、MMP-9、MMP-13、β-catenin蛋白表达。分离正常SD大鼠软骨细胞,IL-1β（10 ng·mL^-1）干预诱导体外OA炎症模型并进行药物干预。番红O染色检测软骨细胞表型变化,ELISA检测软骨细胞IL-6、IL-8含量,免疫荧光染色检测软骨细胞β-catenin表达,qRT-PCR检测软骨细胞中IL-6、IL-8、MMP-3、MMP-9和MMP-13的mRNA表达,Western blotting检测软骨细胞中MMP-3、MMP-9、MMP-13、β-catenin蛋白表达。结果 药物处理对OA导致的软骨组织损伤有不同程度的修复作用;杜仲-当归可下调血清中IL-6、IL-8水平,下调软骨组织中MMP-3、MMP-9、MMP-13、b-catenin的蛋白表达。与IL-1b组相比,杜仲-当归可提高软骨细胞存活率,改善细胞形态,下调软骨细胞IL-6、IL-8水平,降低软骨细胞中b-catenin荧光强度,并下调软骨细胞中IL-6、IL-8、MMP-3、MMP-9、MMP-13的mRNA表达和MMP-3、MMP-9、MMP-13、b-catenin的蛋白表达。结论 体内、体外试验表明补肾活血中药“杜仲-当归”可以通过促进软骨细胞增殖、调控炎症因子水平、改善软骨基质降解、改善软骨病理损伤起到对OA的防治作用,且作用机制可能与介导Wnt/β-catenin信号转导调控MMP-13表达有关。     

盐炒杜仲和杜仲炭的药理对比实验研究

目的:观察盐炒杜仲和杜仲炭对小鼠单核-巨噬细胞、血压、中枢镇静作用的影响,以区别两种炮制品药理作用,为临床应用作为参考。方法:选取100只小鼠随机分为空白对照组、2.0g·kg-1盐炒杜仲醇煎液组、8.0 g·kg-1盐炒杜仲醇煎液组、2.0g·kg-1杜仲炭醇煎液组、8.0 g·kg-1杜仲炭醇煎液组,观察不同剂量的盐炒杜仲和杜仲炭对小鼠单核-巨噬细胞的影响。选取100只小鼠随机分为空白对照组、3.0 g·kg-1盐炒杜仲醇煎液组、9.0 g·kg-1盐炒杜仲醇煎液组、3.0 g·kg-1杜仲炭醇煎液组、9.0 g·kg-1杜仲炭醇煎液组,观察不同剂量的盐炒杜仲和杜仲炭对小鼠血压的影响。选取100只小鼠随机分为空白对照组、5.0g·kg-1盐炒杜仲醇煎液组、10.0 g·kg-1盐炒杜仲醇煎液组、5.0 g·kg-1杜仲炭醇煎液组、10.0 g·kg-1杜仲炭醇煎液组,观察不同剂量的盐炒杜仲和杜仲炭对小鼠中枢镇静作用的影响。结果:与空白对照组比较,高剂量的盐炒杜仲醇煎液组和杜仲炭醇煎液组均显著升高(P<0.05),但杜仲炭醇煎液组的免疫效果更加明显。与空白对照组比较,高剂量的盐炒杜仲醇煎液组和杜仲炭醇煎液组均显著降低小鼠的收缩压和舒张压(P<0.05),但杜仲炭醇煎液组降压效果更加明显。与空白对照组比较,高剂量的盐炒杜仲醇煎液组和杜仲炭醇煎液组均显著促进小鼠睡眠(P<0.05),但杜仲炭醇煎液组镇静效果更加显著。结论:盐炒杜仲和杜仲炭均具有较好的免疫作用、较强的降血压作用和中枢镇静作用,但杜仲炭的药理活性更强。文献引用:李轩.盐炒杜仲和杜仲炭的药理对比实验研究[J].中医学报,2015,30(2):238-240.     

Human splanchnic metabolism during diabetic ketoacidosis.

Splanchnic exchange rates of glucose, acetoacetate, beta-hydroxybutyrate, lactate, pyruvate, glycerol, alanine, glutamine, glutamate, free fatty acids, and triglycerides were measured in eight patients during moderate to severe diabetic ketoacidosis. Their arterial glucose concentration was 20.68 (9.80-52.79) mumole/liter and tic glucose release was 0.77 (0.09-2.44) mmole/min. Gluconeogenesis accounted for about one-half of net splanchnic glucose release, assuming quantitative conversion of net splanchnic extracted lactate, pyruvate, glycerol, alanine, and alpha-ketoglutarate equivalents to glucose. Net splanchnic free fatty acid extraction was 0.24 (0.09-0.52) mmole/min. There was a positive correlation between free fatty acid uptake and ketone-body release. Net splanchnic acetoacetate release was 0.50 (0.05-0.92) mmole/min and beta-hydroxybutyrate release was 0.35 (-0.16 to 0.84) mmole/min. Total ketone-body release was 0.84 (0.37-1.61) mmole/min. The wide ranges of net splanchnic glucose and ketone-body production rates show the heterogeneous characteristics of the diabetic patient in ketoacidosis. It is concluded that the hyperglycemia and hyperketonemia of diabetic ketoacidosis is due to the lack of reciprocity among rates of hepatic glycogenlysis, gluconeogenesis, and ketogenesis resulting in inappropriate net splanchnic release of glucose and ketone bodies.     

杜仲叶绿原酸的提取及精制

对杜仲叶绿原酸的提取进行研究。方法 采用乙醇为溶剂，经正交实验方法对杜仲叶绿原酸的提取进行研究，并采用大孔树脂对其精制。结果 乙醇提取杜仲叶绿原酸的最佳工艺条件为：温度80℃．料液比1：10．乙醇浓度60％，提取时间2小时。所筛选的NKA-Ⅱ树脂是吸附绿原酸的理想吸附剂。结论 适宜于产业化生产。     

杜仲不同提取物对帕金森病小鼠的治疗作用及谱效关系研究

目的探究杜仲不同提取物对帕金森病小鼠的治疗作用及其超高效液相色谱(UPLC)分析与治疗帕金森病的谱效关系。方法通过小鼠爬杆实验、脑内纹状体多巴胺(DA)含量,观察杜仲不同梯度的乙醇提取物对帕金森病小鼠的治疗作用;采用超高效液相色谱-四级杆飞行时间质谱(UPLC-Q-TOF/MS)对杜仲不同提取物进行分析;结合爬杆实验及DA水平结果,采用偏最小二乘法回归(PLSR)分析建立其谱效关系,明确杜仲治疗帕金森病的药效成分。结果杜仲50%、75%乙醇提取物均能明显缩短小鼠爬杆时间;杜仲75%乙醇提取物能显著增加小鼠脑内纹状体DA水平;PLSR分析结果显示,杜仲中杜仲醇苷、鹅掌楸苷、5-羟甲基糠醛、咖啡酸与爬杆实验结果和纹状体多巴胺含量密切相关。结论杜仲乙醇提取物具有抗帕金森病作用,其中75%乙醇提取物效果最显著;杜仲醇苷、鹅掌楸苷、5-羟甲基糠醛、咖啡酸可能是杜仲治疗帕金森病的主要有效成分。     

健脾益肾清热利湿方辅助治疗急性肾盂肾炎的疗效观察

目的：观察健脾益肾清热利湿方辅助治疗急性肾盂肾炎的临床疗效。方法将52例急性肾盂肾炎患者随机分为治疗组和对照组各26例。对照组给予抗生素治疗,治疗组在对照组的基础上给予健脾益肾清热利湿方辅助治疗,两组疗程均为2周。观察两组治疗前后主要临床症状、体征的变化。结果在疗效方面,治疗组总有效率为96.2%,对照组总有效率为80.8%,治疗组疗效优于对照组（P<0.05）;在发热、腰痛、尿路刺激征消除时间方面治疗组明显快于对照组（P<0.01）。结论健脾益肾清热利湿方辅助治疗急性肾盂肾炎效果显著,可明显改善患者的临床症状,并可缩短临床症状的消除时间,能提高治疗的有效率。