Inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.on human carboxylesterases

OBJECTIVE The inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.(TWHF)(celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine)on human carboxylester⁃ase 1(CES1)and CES2 was detected to investigate the herb-drug interactions(HDIs)of TWHF.METHODS Human liver microsomes catalysed hydrolysis of 2-(2-benzoyl-3-methoxyphenyl)benzothi⁃azole(BMBT)and fluorescein diacetate(FD)were used as the probe reaction to phenotype the activity of CES1 and CES2,respectively.The residual activities of CES1 and CES2 were detected by ultrahigh performance liquid chromatography(UPLC)after intervention with celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine(100μmol·L^(-1)).Kinetics analysis,involving half inhibitory concentra⁃tion(IC_(50)),inhibition type and kinetic parameter(Ki),and in vitro-in vivo extrapolation(IVIVE),was carried out to predict the HDIs between these compounds and CES-metabolizing drugs.Molecular docking was performed to analyze the ligand-enzyme interaction.RESULTS Out of the six main con⁃stituents of TWHF,only celastrol exhibited strong inhibition towards both CES1 and CES2,with the inhibitory rates of 97.45%(P<0.05)and 95.62%(P<0.05),respectively.The IC_(50)was 9.95 and 4.02 mol·L^(-1),respectively,and the types of inhibition were all non-competitive inhibition.Based on the kinetics analysis,the Ki values were calculated to be 5.10 and 10.55μmol·L^(-1)for the inhibition of celastrol on CES1 and CES2,respectively.IVIVE indicated that celastrol might disturb the metabolic hydrolysis of clinical drugs in vivo by inhibiting CES1.Molecular docking results showed that hydrogen bonds and hydrophobic contacts contributed to the interaction of celastrol and CESs.CONCLUSION The inhibitory effect of celastrol on CES1 and CES2 might cause HDIs with clinical drugs hydrolysed by CESs.

HISTOCHEMICAL EFFECT OF THE MULTIGLYCOSIDES OF TRIPTERYGIUM WILFORDII HOOK.F.(GTW)ON THE TESTIS AND EPIDIDYMIS OF RATS

SD Twenty adult male rats were equally divided into a contro1 and an experimcntal group.GTW,10 mg/kg per day,was given to the experimental rats through gastric gavage 6 days a week for 8 weeks.To the control rats,the same amount of vehicle was given The animals were sacrificed when they became infertile and the fol1owing parameters of the testis and epididymis were examined:l)DNA(Feulgen′s method),2)RNA(Brachet′s tech-uique),3)LDH-X(Lojda′s method),4)ATPase(Wachstein′s procedure),5)Succinate dehydrc-genase(SDH,Pearson's method),6)Non-specific esterase(NSE,Lojda's technique)and 7)PAS reaction.Routine examination of the epididymal spermatozoa was also carried out.

比较蛋白质组学揭示茉莉酸甲酯诱导的雷公藤甲素生物合成

目的:探索雷公藤甲素生物合成途径,以提高其产量或用于合成生物学异源生产。方法:使用无标记(lable-free)比较蛋白质组学对茉莉酸甲酯(MeJA)诱导的雷公藤悬浮细胞进行测序,通过京都基因与基因组百科全书(KEGG)、基因本体(GO)分析等方法,结合基因转录表达分析和基因体内功能研究,逐步筛选参与雷公藤甲素生物合成的功能基因。结果:MeJA诱导不同时间的样品组鉴定得到376个差异蛋白质,包括8个细胞色素P450酶(CYP450)、3个转录因子(TF)和2-氧戊二酸依赖的双加氧酶(2ODD)。其中CYP71BE89、CYP82AQ2和Tw2ODD基因在MeJA诱导及植物不同组织部位中,均与已知参与雷公藤甲素母核环化的二萜合酶TwTPS7(v2)和TwTPS27(v2)基因具有相似的表达模式,表明其可能与雷公藤甲素生物合成相关。进一步利用植物细胞体内功能研究,验证了CYP71BE89、CYP82AQ2和Tw2ODD基因干扰使雷公藤甲素积累分别降低了24.1%、41.4%和71.4%,表明了3个基因与雷公藤甲素生物合成直接相关。结论:利用比较蛋白质组学技术揭示了几种与雷公藤甲素生物合成相关的蛋白,进一步表征了雷公藤甲素生物合成与外源激素刺激之间的关系,并为发掘中药活性成分生物合成途径提供了一种有效方法。

雷公藤血液毒性机制及防治的研究进展

雷公藤用药历史悠久,具有祛风除湿、活血化瘀、清热解毒等功效,常用于多种免疫系统疾病的治疗,疗效显著。作为一种传统有毒中药,其血液毒性也不容小觑,限制了其在很多领域的应用,也愈发受到广泛关注。通过综述雷公藤相关研究,梳理分析血液毒性表现特点发现,患者临床表现轻重不同,重者可能会产生骨髓抑制,但发生率低,且大多可逆。由于其所含化学成分复杂,不同成分致血液毒性产生的机制各异。对雷公藤所含有的主要毒性成分进行归类梳理,并从抑制细胞周期、影响转录调节、致突变及诱导细胞凋亡等方面总结其血液毒性作用机制,对雷公藤减毒的中西医防治措施进行整理与发掘,总结单味中药、中药复方及西药减毒机制,为其在临床的使用提供参考与依据。

内生真菌对雷公藤组培苗生长、生理及药用成分含量的影响

分析了雷公藤(Tripterygium wilfordii Hook.f.)11株内生真菌的促生潜力,并将11株内生真菌与雷公藤组培苗共生培养,研究了其对组培苗生长、叶片叶绿素含量和叶绿素荧光参数、主要营养元素含量以及雷公藤红素和雷公藤甲素含量的影响。结果表明:雷公藤11株内生真菌均具有分泌吲哚乙酸、产铁载体和溶磷能力,其中,菌株NS6、NS7、NS14和NS32的吲哚乙酸分泌量较大(大于17.00 mg·L^(-1)),菌株NS1、NS4、NS6、NS25和NS33的产铁载体能力很高〔80%<铁载体相对产量(SP)≤100%〕,菌株NS1、NS4、NS6、NS13和NS31溶磷量较大(大于0.70 mg·L^(-1))。与对照相比,菌株NS1、NS6、NS31、NS32和NS33总体显著(P<0.05)增加雷公藤组培苗的单株鲜质量和干质量,其中,菌株NS6、NS32和NS33显著增加组培苗株高,菌株NS1和NS6显著促进组培苗最长根长,菌株NS6显著增加组培苗单株叶片数。与对照相比,除菌株NS13外,其余10株菌株均显著提高组培苗叶片叶绿素含量;菌株NS1、NS7、NS18、NS31和NS32显著提高组培苗叶片最大荧光,菌株NS14显著提高组培苗叶片PSⅡ最大光化学效率。与对照相比,菌株NS1、NS7、NS14、NS31、NS32和NS33显著提高组培苗C、P和K含量,菌株NS1、NS4、NS32和NS33显著提高组培苗N含量。与对照相比,11株内生真菌均能显著提高组培苗根和茎中雷公藤红素含量,仅菌株NS33显著提高叶片中雷公藤红素含量;与11株内生真菌共生培养的组培苗茎和叶片中未检出雷公藤甲素,菌株NS6、NS7、NS13、NS25、NS32和NS33显著提高根中雷公藤甲素含量。综合研究认为,菌株NS1、NS6、NS31、NS32和NS33具有多种促生潜力且能改善雷公藤组培苗生长,促进根和茎中雷公藤红素积累;菌株NS6、NS32和NS33促进根中雷公藤甲素积累,具有较好的应用潜力。

基于Citespace知识图谱雷公藤治疗类风湿性关节炎研究热点与趋势分析

目的:通过分析雷公藤治疗类风湿性关节炎的相关英文文献,归纳该领域的研究热点,总结发展趋势,为后续雷公藤治疗类风湿性关节炎的进一步研究提供参考。方法:以Web of Science、Wiley Online Library和Embase数据库,基于CiteSpace V 5.8.R3软件对2000-2021年雷公藤与类风湿性关机炎相关的英文文献进行文献计量学分析,从机构发文量、作者发文量、关键词以及发展趋势等研究方面进行可视化分析。结果:研究纳入252篇英文文献,2017-2021年发文量占比35.3%,主要集中在亚洲国家及地区,包括中国、日本和中国台湾,而《民族药理学杂志》是发文量最多的英文期刊,发文量最多的作者是以Aiping Lu、Cheng Lu、Cheng Xiao等为代表的团队,研究内容是雷公藤治疗类风湿性关节炎的基础研究及临床应用,共计发文26篇,发文机构主要集中在国内高校,其中中国中医研究院是发文量最多的研究机构,多达42篇,并形成以中国中医科学院、中国药科大学和北京中医药大学为中心的主要发文研究机构。关键词可视化分析显示研究方向主要集中在甲氨蝶呤联合用药、活化作用、定量分析等方面,而未来研究热点主要集中在信号通路、机制研究等方面。结论:雷公藤和类风湿性关节炎相关英文文献发文量逐年上升,预测未来研究热点和发展方向主要集中于调控信号通路和挖掘治疗疾病的基础机制研究方面。目前,雷公藤治疗类风湿性关节炎的研究仍处在发展深入阶段,而相关研究团队应该进一步加强国际间的合作,促进后续该领域研究的发展,并提供参考依据。

In vitro propagation of a medicinal plant: Tripterygium wilfordii Hook f.

In this study a reliable protocol was developed for the establishment of commercial in vitro cultures of Tripterygium wilfordii Hook f.. Juvenile shoots from one-year-old elite plants were used as the source of explants. New axillary shoots were obtained after 30 days of culture on a MS medium supplemented with BAP （2.0 mg.L^-1） and NAA （0.1 mg.L^-1）. The optimal multiplication medium was a modified MS medium supplemented with BAP （1.0 mg.L^-1） and NAA （0.1 mg.L^-1）. This yielded a multiplication rate of 2.4 for each subculture. Slightly more than 92% of shoots rooted when cultured on a modified MS medium containing IBA （0.2 mg.L^-1） and activated charcoal （0.5 mg.L^-1）. Activated charcoal promoted both a strong and a high rooting rate during the rooting phase. Plantlets were transferred to pots for a short acclimatization stage in a greenhouse where 95% of the plantlets survived. This highly reproducible procedure can be adopted for large-scale propagation of T. wilfordii.

Effect of Three Extraction Methods on the Volatile Component of Illicium verum Hook.f.Analyzed by GC-MS

The physical properties and chemical components of three oil samples extracted from Illicium verum Hook. f. by steam distillation （SD）, solvent extraction （SE） and supercritical fluid extraction （SFE） were compared with one another and analyzed by Gas Chromatography-Mass Spectrometry （GC-MS）. The quality parameters of star anise essential oil from SFE were close to that of those came from SD and SE. Although the extraction yield of star anise by SFE （9.2 %） was close to the value from SE （9.3%）, it was yet higher than that came from SD （8.2%）. For sensory evaluation, however, three oils were significantly different. The odor and taste of the products from SFE and SE were generally more natural and vivid than that came from distilled oil. The volatile compound revealed that significant differences of the composition existed in the distilled oil and the oleoresins prepared by SFE and SE.

Isolation and Purification of Triptolide from the Leaves of Tripterygium wilfordii Hook F

Triptolide is an important active component of Tripterygium wilfordii Hook F (TWHF) and possesses anti-inflammatory, immunosuppressive, male anti-fertility, and anticancer properties. A new method combining different techniques, including solid-liquid extraction, liquid-liquid partition, column chromatography and high-speed counter-current chromatography (HSCCC) but avoiding the use of chloroform, was developed for the isolation and purification of triptolide from the leaves of TWHF. 48 mg of triptolide at 96.5% purity was obtained from 1 kg of air-dried leaves of TWHF.

Effect of the Multiglycoside of Tripterygium Wilfordii Hook f. (Tii) on Cornea Allograft Rejection Model in Rabbit

Purpose:Toexamine the effect of Tii treatment of cornea graft survival in a rab-bit model.Methods:Tii was administrated orally after eccentrical corneal transplantation.Survival times were determined by biomicroscopy.Cytotoxic T lymphocytes(CTL)and delayed-type hypersensitivity(DTH)responses to donor alloantigens were assessed at ady 16after heterotopic corneal grafts.Results:Administration of Tii reduced the incidence and prologed the graft sur-vival time.Both CTLand DTH responses to donor alloantigens were severely ed-pressed in hosts treated with Tii.However,combination of Tii and cyclosporine further enhanced the immunosuppressive effects described above.Conclusions:Tii is a potent immunosuppressant with the ability to prolong allo-graft survival in the rabbit penetrating keratoplasty model and may have coordi-native effects with CsA through different mechanisms.Further studies are neces-sary to define any potentially coordinative role in the prevention of allograft rejec-tion in human keratoplasty.Eye Science 1995;11:168-172.

A Novel Diterpenoid,11-O-β-D-glucopyranosylneo-triptophenolide, from Tripterygium wilfordii Hook. f

A new diterpenoid was isolated from the ethanolic extract of the dried root bark of Tripterygium wilfordii Hook.f. It is the first example of abietane diterpenoid glycoside isolated from Tripterygium wilfordii Hook.f. Its structure was identified to be 11-O-b-D-glucopyranosyl- neotriptophenolide based on spectral methods.

Safety Evaluation of a Compound Lonicera rupicola Hook.f. et & Thomson Injection

Safety of a compound Lonicera rupicola Hook. f. et & Thomson injection was evaluated by local irritation experiments including conjunctiva,skin and muscle irritant experiments,and the effect on weight gain was studied. The results showed that the compound L. rupicola Hook. f. et & Thomson injection had no irritation to rabbit eyes and weak irritation to rabbit ears,and had relatively strong irritation to the leg muscles of the mice. Its metabolism in mice had no abnormal toxicity.

基于足细胞保护探讨雷公藤治疗糖尿病肾病研究进展

糖尿病肾病(DKD)是终末期肾病的主要原因。DKD早期症状表现为尿蛋白增加,与足细胞功能和形态改变有关,表明足细胞损伤是DKD进展的重要因素。雷公藤具有显著的抗炎,抗氧化和免疫调节等药理活性。本文从抑制足细胞脱落、凋亡、氧化应激和炎症反应,改善足细胞转分化和稳定细胞骨架等方面探讨雷公藤对DKD足细胞的保护作用,为更好地防治DKD提供研究策略。

藤苦参组培快繁技术

以藤苦参(Streptocaulon griffithii Hook.f.)种子为试验材料,在种子繁殖基础上开展组培繁殖,探索不同植物生长调节剂对藤苦参无菌苗不定芽增殖、生根等的影响。结果表明,最适合藤苦参种子萌发的培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA+1 g/L活性炭或MS+1.5 mg/L 6-BA+0.5 mg/L NAA+1 g/L活性炭;适合藤苦参丛生芽分化的培养基为MS+3.0 mg/L 6-BA+0.5 mg/L NAA。以种子为外植体,筛选其组织培养各阶段培养基配方,建立藤苦参组培快繁技术,以期为藤苦参资源保护利用奠定基础。

青藏高原种植业碳排放量测度——以青海省青稞种植业为例

为研究青藏高原种植业碳排放量测度问题,以青海省青稞(Hordeum vulgare L.var.nudum Hook.f.)种植业产生的碳排放量、碳排放强度、碳成本为测算目标进行测定。通过对青海省1997—2019年的碳排放总量和农业碳排放量现状进行分析,发现青海省碳排放总量仍居高不下,农业碳排放量也始终处于较高水平。基于1997—2019年青海省农业碳排放数据,使用Matlab软件中GUI工具箱搭建非线性次回归神经网络模型,并使用神经网络时序工具对青海省农业2020—2026年碳排放进行多步预测,结果显示2020—2026年青海省农业碳排放量仍呈高位波动增长的趋势,迫切需要开展“双碳”减排。依据青稞种植业碳排放来源,建立了农用柴油、人工、化肥、农药、农膜、N2O排放6个测算指标。并基于2015—2020年各测算指标的数据建立IPCC清单估算模型,最终测得青海省青稞种植业2015—2020年碳排放总量、碳排放强度和碳成本,结果显示,2015—2020年青海省青稞种植业碳排放总量并未随着青稞种植面积的增大而出现大幅度增长,而青稞种植业碳排放强度和碳成本随着青稞种植面积的增大而有所降低。

雷公藤属药用植物的研究进展

雷公藤属药物植物具有抗炎、抗免疫、抗肿瘤、抗H IV等多种药理活性。临床上广泛用于治疗类风湿性关节炎、慢性肾炎等自身免疫性疾病。其化学成分复杂,主要包括倍半萜类(包括倍半萜生物碱)、二萜类、三萜类和少量木质素等。综述近年来雷公藤属药用植物化学成分、药理作用等方面的研究成果,指出雷公藤类药物具有广阔的发展前景。

雷公藤属3种植物不同群体和个体中雷公藤甲素的研究

目的测定雷公藤属3种植物(雷公藤、昆明山海棠和黑蔓)不同群体和个体中雷公藤甲素(tripto lide),为评价雷公藤药材质量和寻找雷公藤的优质种质奠定基础。方法建立雷公藤甲素HPLC测定方法,并测定了全国主要分布区25个群体91个个体的木质部和韧皮部中的雷公藤甲素。结果黑蔓的雷公藤甲素质量分数很低;雷公藤和昆明山海棠种间的雷公藤甲素差异不明显,个体质量分数木质部为1.0×10-6~5.83×10-5;韧皮部为2.3×10-6~1.030×1-0 4。结论雷公藤和昆明山海棠不同个体雷公藤甲素质量分数最高值与最低值相差约50倍,不同居群雷公藤甲素质量分数最高值与最低值相差10多倍,不同来源的药材质量差异极大,严重影响用药的安全性;雷公藤甲素质量分数高的居群位于浙江西南部和中部、湖南新宁、贵州雷山和安徽黄山;湖南新宁、贵州雷山和浙江江山居群中有雷公藤甲素质量分数极高的个体,值得进一步研究,以寻找质量分数高的优良单株。

雷公藤组培产物中雷公藤甲素和总生物碱含量的测定

建立了同一提取物中高效液相色谱法测定雷公藤甲素和紫外分光光度法测定雷公藤总生物碱含量的方法,样品用乙酸乙酯超声提取、过中性氧化铝柱,净化后用45%甲醇溶解,测定雷公藤的根皮、叶、雷公藤组培产物及培养液中雷公藤甲素和雷公藤总生物碱的含量。结果表明,雷公藤甲素的检测范围为1～100μg/mL(R2=0.9999);雷公藤总生物碱在5～100μg/mL范围内线性关系良好(R2=0.9998);在同一提取液中,雷公藤甲素和雷公藤总生物碱的添加回收率分别为100.5%(RSD=0.63%)和99.8%(RSD=1.22%)。研究结果表明,根愈伤组织中雷公藤甲素含量为根皮粉中的1.25倍,培养液中雷公藤甲素产量占56%,总生物碱产量占65%。每生产1g不定根培养物,雷公藤甲素产量为根皮粉中的10倍,雷公藤总生物碱产量为根皮粉中的2倍。

HPLC法测定雷公藤及其片剂中雷公藤红素

目的建立雷公藤及其片剂中雷公藤红素的HPLC测定方法.方法 HPLC外标法.Agilent Zorbax SC-C18柱为色谱柱,甲醇-1%醋酸溶液(87:13)为流动相,检测波长为425 nm,体积流量为1.0 mL/min.结果雷公藤红素线性范围为40.96～204.8μg/mL(r=0.999 6),药材的平均回收率为98.37%,RSD为1.01%(n=9),制剂的平均回收率为98.59%,RSD为1.18%(n=9).结论本方法操作简便、准确,可作为雷公藤类药物中雷公藤红素的定量分析方法.

雷公藤无性系苗木光合生理特性研究

对雷公藤(Tripterygium wilfordiiHook.f.)23个无性系苗木的光合生理特性进行研究。结果表明:雷公藤苗木净光合速率Pn日变化曲线为不对称的双峰曲线,蒸腾速率tr日变化曲线为单峰曲线,气孔导度Gs、胞间CO2浓度Ci的日变化分别呈勺形、倒双峰变化趋势;净光合速率随光合有效辐射增加而达到峰值,随后反而降低,光补偿点为17μmol.m-.2s-1,光饱和点为600μmol.m-.2s-1;净光合速率与气温、叶温、光合有效辐射、蒸腾速率呈极显著正相关关系,与大气相对湿度、叶内湿度、胞内CO2浓度、胞间CO2浓度呈极显著负相关关系;通过分析比较,初选获得编号为17、13、16、10、11、4、6共7个具有较高净光合速率的优良无性系。