Protective effects of Dragon's Blood on blood coagulation and NO/iNOS level in myocardium and serum of rats in simulated microgravity

To investigate effects of Dragon＇s Blood（DB）,a traditional Chinese medicine,on blood coagulation and NO/iNOS level in myocardium and serum of rats in simulated microgravity for the first time,Sprague Dawley（SD）rats were randomly divided into six groups：（a）5-day control group,（b）5-day model group,（c）5-day drug group,（d）21-day control group,（e）21-day model group,and（f）21-day drug group.Blood coagulation and NO/iNOS level in myocardium and serum were examined after 5 and 21 days of simulated microgravity respectively.The results showed that blood of tail-suspended rats was in a hypercoagulable state that could not be converted with time extending.Conversely,DB changed these parameters towards normal level and the curative effects became better when tail-suspension lasted till the 21 st day.NO concentration of both myocardium and serum for two periods all increased markedly and DB could effectively reduce these increases except that of 21-day myocardium NO.Activity of iNOS increased markedly as early as 5 days and became more serious on the 21 st day,while DB showed preventive effect on the 21 st day.Western Blot analysis illustrated that the expression of iNOS in the 5-day model group increased significantly over the 5-day control group and the expression in the 5-day drug group dramatically returned to the normal level.The similar trend was observed on the 21-day groups without notable variances.The findings of this study can serve for the further use of Dragon＇s Blood in space diseases.

Material basis for inhibition of Dragon's Blood on evoked discharges of wide dynamic range neurons in spinal dorsal horn of rats

In vivo experiments were designed to verify the analgesic effect of Dragon’s Blood and the material basis for this effect. Extracellular microelectrode recordings were used to observe the effects of Dragon’s Blood and various combinations of the three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from Dragon’s Blood on the discharge activities of wide dynamic range (WDR) neurons in spinal dorsal horn (SDH) of intact male Wistar rats evoked by electric stimulation at sciatic nerve. When the Hill's coefficients describing the dose-response relations of drugs were dif-ferent, based on the concept of dose equivalence, the equations of additivity surfaces which can be applied to assess the interaction between three drugs were derived. Adopting the equations and Tal-larida's isobole equations used to assess the interaction between two drugs with dissimilar dose-response relations, the effects produced by various combinations of the three components in modulating the evoked discharge activities of WDR neurons were evaluated. Results showed that Dragon’s Blood and its three components could inhibit the evoked discharge frequencies of WDR neurons in a concentration-dependent way. The Hill's coefficients describing dose-response relations of three components were different. Only the combined effect of cochinchinenin A, cochinchinenin B and loureirin B was similar to that of Dragons Blood. Furthermore, the combined effect was synergistic. This investigation demonstrated that through the synergistic interaction of the three components Dragon’s Blood could interfere with the transmission and processing of pain signals in spinal dorsal horn. All these further proved that the combination of cochinchinenin A, cochinchinenin B, and loureirin B was the material basis for the analgesic effect of Dragon’s Blood.

A botanical medicine dragon’s blood exhibited clinical antithrombosis efficacy similar to low molecular weight heparin

Deep vein thrombosis(DVT)is a common complication following traumatic fracture with a 0.5%–1%annual incidence.Low molecular weight heparin(LMWH)is the most commonly used anticoagulation drug for DVT prevention,but treatment with LMWH is invasive.Our aim is to compare the antithrombotic effect of dragon’s blood,an oral botanical anticoagulant medicine approved by the Chinese FDA,with LMWH in patients undergoing hip fracture surgery and to explore the molecular mechanisms of anticoagulation treatment.Our study recruited patients and divided them into LMWH and dragon’s blood treatment group.Coagulation index tests,Doppler ultrasound and mRNA sequencing were performed before and after anticoagulation therapy.There was no significant difference in postoperative DVT incidence between the two groups(23.1%versus 15.4%,P=0.694).D-dimer(D-D)and fibrinogen degradation product(FDP)showed significant reductions in both groups after anticoagulation treatments.We identified SLC4A1,PROS1,PRKAR2B and seven other genes as being differentially expressed during anticoagulation therapy in both groups.Genes correlated with coagulation indexes were also identified.Dragon’s blood and LMWH showed similar effects on DVT and produced similar gene expression changes in patients undergoing hip fracture surgery,indicating that dragon’s blood is a more convenient antithrombosis medicine(oral)than LMWH(hypodermic injection).

Modulation of dragon's blood on tetrodotoxin-resistant sodium currents in dorsal root ganglion neurons and identification of its material basis for efficacy

To clarify the modulation of dragon's blood on the tetrodotoxin-resistant (TTX-R) sodium currents in dorsal root ganglion (DRG) neurons and explore its corresponding material basis for the efficacy, using whole-cell patch clamp technique, the effects of dragon's blood and the combined effects of three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from dragon's blood on the TTX-R sodium currents in acute-isolated DRG neurons of rats were observed. According to the operational definition of material basis for the efficacy of TCM established, the material basis of the modulation on the TTX-R sodium currents in DRG neurons of dragon's blood was judged from the experimental results. The drug interaction equation of Greco et al. was used to assess the interaction of the three components extracted from dragon's blood. This investigation demonstrated that dragon's blood suppressed the peak TTX-R sodium currents in a dose-dependent way and affected the activations of TTX-R sodium currents. The effects of the combination of cochinchinenin A, cochinchinenin B, and loureirin B were in good agreement with those of dragon's blood. Although the three components used alone could modulate TTX-R sodium currents, the concentrations of the three components used alone were respectively higher than those used in combination when the inhibition rates on the TTX-R sodium currents of them used alone and in combination were the same. The combined effects of the three components were synergistic. These results suggested that the interference with pain messages caused by the modulation of dragon's blood on TTX-R sodium currents in DRG neurons may explain some of the analgesic effect of dragon's blood and the corresponding material basis for the efficacy is the combination of cochinchinenin A, cochinchinenin B, and loureirin B.

A New Phenylpropanoid Glycoside from Dragon＇s Blood of Dracaena cambodiana

A new phenylpropanoid glycoside, named cambodianin F（1）, together with three known compounds, 1-O-β-D-glucopyranosyl-2-hydroxy-4-allylbenzene（ 2 ）, 1-O-（ 6-O-a-L-rhamnopyranosyl-β-D-glucopyranosyl）-2-hydroxy- 4-allylbenzene（3） and 1,2-di-O-β-D-glucopyranosyl-4-allylbenzene（4） was isolated from the dragon＇s blood of Dracaena cambodiana. The new compound was elucidated by HR-ESI-MS and spectroscopic techniques（UV, IR, 1D and 2D NMR）.

A chromosome-level genome assembly for Dracaena cochinchinensis reveals the molecular basis of its longevity and formation of dragon’s blood

Dracaena,a remarkably long-lived and slowly maturing species of plant,is world famous for its ability to produce dragon’s blood,a precious traditional medicine used by different cultures since ancient times.However,there is no detailed and high-quality genome available for this species at present;thus,the molecular mechanisms that underlie its important traits are largely unknown.These factors seriously limit the protection and regeneration of this rare and endangered plant resource.Here,we sequenced and assembled the genome of Dracaena cochinchinensis at the chromosome level.The D.cochinchinensis genome covers 1.21 Gb with a scaffold N50 of 50.06 Mb and encodes 31619 predicted protein-coding genes.Analysis showed that D.cochinchinensis has undergone two whole-genome duplications and two bursts of long terminal repeat insertions.The expansion of two gene classes,cis-zeatin O-glucosyltransferase and small auxin upregulated RNA,were found to account for its longevity and slow growth.Two transcription factors(bHLH and MYB)were found to be core regulators of the flavonoid biosynthesis pathway,and reactive oxygen species were identified as the specific signaling molecules responsible for the injuryinduced formation of dragon’s blood.Our study provides high-quality genomic information relating to D.cochinchinensis and significant insight into the molecular mechanisms responsible for its longevity and formation of dragon’s blood.These findings will facilitate resource protection and sustainable utilization of Dracaena.

龙血竭高效液相色谱特征研究

目的:用HPLC法对龙血竭指纹图谱进行研究。方法:采用色谱条件Eclipse XDB-ODS柱(4.6 mm×150 mm,5μm);流动相乙腈-水,梯度洗脱进行色谱分离;检测波长为275 nm;流速为1.0 mL.min-1;柱温为40℃,以龙血素B作为参照物。结果:初步建立了龙血竭的HPLC指纹图谱,并进行相似度评价。结论:方法简单可行,能够有效地控制龙血竭的内在质量。

龙血素B在模拟失重大鼠体内的血浆药物动力学研究

目的研究模拟失重大鼠灌胃龙血竭后有效成分龙血素B的血浆药物动力学。方法采用3周尾吊大鼠模型模拟失重状态,单次灌胃给予10.6 g/kg龙血竭后,收集各个时间点血浆样本,HPLC-MS/MS法测定尾吊大鼠血浆中龙血素B的含量,并求算药物动力学参数。结果龙血素B在模拟失重大鼠体内血药浓度-时间曲线下面积(AUC0-T)为(23.4±12.4)μg/L·h,达峰时间(Tmax)为(0.33±0.11)h,达峰浓度Cmax(2.77±0.71)μg/L,表观分布容积(Vc)为(677.6±111.6)L/kg。生物半衰期(T0.5)为(8.47±1.7)h。结论本文首次报道了龙血素B在模拟失重大鼠体内的药物动力学参数,为龙血竭对抗空间疾病的相关研究提供了药动学数据。

血竭提取物对角质形成细胞增殖的影响

目的:观察血竭提取物对体外培养角质形成细胞增殖的影响,探讨血竭促进创面愈合的机制。方法:将血竭经过氯仿、乙酸乙酯、乙醇回流提取得到三种提取液,进行角质形成细胞的培养,噻唑蓝(MTT)法检测不同血竭提取物在不同浓度以及不同时间点对体外培养角质形成细胞增殖的影响,并绘制最适浓度下细胞生长曲线。利用流式细胞仪分析最适浓度培养条件下角质形成细胞的细胞周期变化。结果:血竭乙酸乙酯提取物在0.0625～0.5mg/ml浓度范围内,促进角质形成细胞增殖,且呈剂量依赖性,在0.5mg/ml浓度值时促进作用最显著,此条件下细胞DNA合成S期较对照组增加25.7%(P<0.01)。结论:血竭乙酸乙酯提取物具有显著促进角质形成细胞增殖作用,提示血竭可能具有促进创面愈合中再上皮化的作用。

自制血竭膏治疗颌面部损伤的疗效观察

目的 观察自制血竭膏对颌面部擦伤、烧伤患者的治疗效果。方法 用自制血竭膏敷贴创口与用凡士林油纱布覆盖创口进行比较。结果 用血竭膏敷贴创口的治疗效果显著好于用凡士林油纱布覆盖 (P <0 0 1)。结论 血竭膏对颌面部擦伤和烧伤有良好治疗效果。

剑叶龙血树根、茎、叶中血竭成分分析

目的通过对比剑叶龙血树根、茎、叶成分与龙血竭成分间的异同及对剑叶龙血树根、茎、叶中龙血素A、B的测定,寻找龙血竭原料的可能来源。方法薄层色谱法及高效液相色谱技术。结果剑叶龙血树根、茎、叶中均含有微量的龙血素A,分别为:58、53、15μg/g;叶中还含有微量的龙血素B,为2μg/g,而根、茎样品中未能检测到龙血素B。结论从色谱峰的积分面积及保留时间来看,剑叶龙血树根、茎、叶成分与血竭原料有少许的相关性,但差异显著;根、茎、叶3者间的化学组成成分也存在着显著差异,剑叶龙血树根、茎、叶中龙血素量都较低,不能直接用作生产血竭的原料。

海南血竭总黄酮对肝纤维化大鼠TGFβ1及Ⅰ、Ⅲ型胶原蛋白表达的影响

目的：探讨海南血竭总黄酮对肝纤维化大鼠Ⅰ、Ⅲ型胶原蛋白沉积及转化生长因子β1（TGFβ1）表达的影响。方法：采用CCl4诱导的大鼠产生肝纤维化模型,设置模型组、空白组、秋水仙碱治疗组、海南血竭总黄酮治疗组,分别灌胃给药。实验末期,取肝,采用HE染色,观察肝脏组织的病理学变化。采用免疫组化（SP）法,RT-PCR法观察TGFβ1的表达。采用免疫组化（SP）法观察I、Ⅲ型胶原蛋白的表达。结果：与正常对照组比较,四氯化碳模型大鼠肝脏出现典型的纤维化表现,肝脏胶原纤维间隙广泛形成,肝小叶与肝窦内胶原增生沉积明显,Ⅰ、Ⅲ型胶原（0.58±0.09）vs（7.40±0.55）,（1.36±0.27）vs（6.48±0.82）,（P均〈0.001）及TGFβ1表达明显增加;大剂量血竭总黄酮应用可以明显减轻肝脏胶原纤维增生沉积（P〈0.05）,抑制肝脏Ⅰ、Ⅲ型胶原蛋白（2.38±0.57）vs（7.40±0.55）,（2.13±0.44）vs（6.48±0.82）,（P〈0.050.01）及TGFβ1表达。结论：海南血竭总黄酮可抑制肝纤维化大鼠肝脏Ⅰ、Ⅲ型胶原及TGFβ1表达,发挥对肝纤维化的抑制作用。

中国血竭基源植物的研究与利用

血竭是我国传统名贵中药,目前商品血竭主要来源于棕榈科黄藤属(Daemonorps)植物的果实和百合科龙血树属(Dracaena)植株茎干的红色树脂。在我国境内发现的、有野生分布的血竭基源植物只为海南龙血树和剑叶龙血树两种。系统综述了我国血竭基源植物的资源状况、血竭形成机理、血竭基源植物的开发利用、人工繁育及栽培技术等方面的研究进展,并对野生血竭基源植物资源的保护、开发与利用、血竭产生机制的研究等进行了展望。

复方血竭贴膏对子宫内膜异位症大鼠血浆PGF2α及血清IL-1β的干预作用

[目的]研究复方血竭贴膏的制备工艺,评价该制剂穴位敷贴对子宫内膜异位症模型大鼠的治疗效果,并进一步探讨可能的作用机理。[方法]①制剂制备:以血竭为主药经过稠膏制备、挥发油提取、浸膏制备制成血竭贴膏;②动物实验:采用了自体子宫内膜移植,建立子宫内膜异位症模型,设立对照,观察各组大鼠血浆PGF2α及血清IL-1β水平,[结果]复方血竭贴膏具有抑制异位内膜生长、能够降低大鼠血浆中PGF2α水平,对IL-1β水平的影响不明显。[结论]复方血竭贴膏穴位敷贴能够抑制异位内膜生长、促进腺体萎缩,从而达到治疗子宫内膜异位症作用。

龙血竭中龙血素A在大鼠体内的药动学研究

目的考察龙血竭中龙血素A在大鼠体内的药动学行为。方法按100 g大鼠给予龙血竭250 mg灌胃给药,测定不同时间点的血药浓度,并以3P97软件计算其药动学参数。结果龙血素A的Ka为(1.46±0.19)h-1,t1/2(Ka)为(0.48±0.06)h,t1/2(K)为(2.59±0.67)h,tmax为(2.00±0.55)h,Cmax为(1.13±0.24)μg.mL-1,AUC为(2.84±0.68)μg.h.mL-1。结论龙血素A在大鼠体内符合线性动力学一室开放模型,吸收较快。

复方血竭治疗溃疡性结肠炎的Meta分析

目的评价复方血竭保留灌肠治疗溃疡性结肠炎的临床治疗效果。方法通过计算机及手工检索国内期刊公开发表的复方血竭制剂保留灌肠治疗溃疡性结肠炎的临床试验资料,采用Jadad质量标准对文献进行评价,利用RevMan 5.1.7.0软件对各研究结果的总体效应进行Meta分析,并进行敏感性分析;用漏斗图分析发表性偏倚。结果 6篇文献符合纳入标准,共678例病例,其中3篇文献质量较好,3篇属于低质量文献。Meta分析结果显示:各独立试验间无明显的异质性,合并效应值OR=3.08,95%CI为(1.29,7.40),z=2.52,P<0.05,复方血竭保留灌肠治疗溃疡性结肠炎的临床疗效优于对照组,差异有统计学意义。敏感性分析显示评价结果比较稳定,漏斗图图形不呈现左右对称,考虑存在一定的发表偏倚。结论复方血竭保留灌肠治疗溃疡性结肠炎有一定疗效。

HPLC法测定纳米龙血竭胶囊中龙血素A的含量和包裹率

目的:建立HPLC法测定纳米龙血竭胶囊中龙血素A的含量和包裹率.方法:在C18柱,甲醇-1%冰乙酸溶液(体积比66∶34)为流动相,检测波长280nm,柱温35°C条件下,HPLC测定样品.结果:龙血素A在0.22~7.78μg范围内有良好线性关系,r=0.997,方法回收率94.42%,RSD为1.94%.结论:此方法可用于测定自制纳米龙血竭胶囊中龙血素A含量和龙血竭的包裹率.

近红外漫反射光谱聚类分析用于血竭的鉴别

建立用近红外漫反射光谱法鉴别血竭的方法,采用聚类分析方法进行分类鉴别,快速、准确地鉴别了不同产地的血竭。近红外漫反射光谱法快速、简便、无损,可用于血竭等中药的分类鉴别。

两种工艺提取的龙血竭体外抗真菌活性比较

【目的】对两种工艺提取的龙血竭进行体外抗真菌试验 ,测定其最低抑菌浓度 (MIC)值。【方法】采用试管液基稀释法进行试验。【结果】测得免加热工艺提取的龙血竭抗白色念珠菌、光滑念珠菌、近平滑念珠菌、热带念珠菌、克柔念珠菌、须癣毛癣菌的MIC50 为 1 2 5 ,5 ,0 313,1 2 5 ,5 ,0 175mg/ml,传统工艺提取的龙血竭的MIC50 为 1 2 5 ,5 ,2 5 ,2 5 ,5 ,1 2 5mg/ml。【结论】免加热工艺提取的龙血竭抗近平滑念珠菌和须癣毛癣菌的作用比传统工艺提取的龙血竭作用强。

龙血竭总黄酮对酸敏感离子通道的调控及其镇痛作用

为研究龙血竭总黄酮(total flavonoids of Dragon′s Blood,TFDB)对背根神经节(dorsal root ganglion,DRG)细胞膜上酸敏感型离子通道(acid-sensing ion channels,ASICs)的影响,分析TFDB通过调控ASICs产生镇痛作用的机制,通过急性分离SD大鼠DRG细胞,用全细胞膜片钳技术,观察记录浓度为0.00001%、0.0001%、0.001%、0.01%和0.1%的TFDB对pH 5.0诱发的ASICs电流的影响;观察TFDB(20 mg·kg^(-1))对冰醋酸致小鼠扭体反应的影响及对完全弗氏佐剂(complete Freund′s adjuvant,CFA)诱导大鼠产生的炎性疼痛的影响;提取完全弗氏佐剂诱导的炎症模型大鼠DRG细胞进行免疫组化实验,观察了TFDB对ASIC3蛋白表达的影响.结果表明:TFDB(20 mg·kg^(-1))可以浓度依赖地抑制ASICs电流它对冰醋酸引起的扭体反应具有显著抑制作用(P<0.001),对完全弗氏佐剂诱导的大鼠炎性痛具有明显镇痛效果(P<0.05),同时可以显著抑制ASIC3蛋白表达(P<0.05).故TFDB能够抑制酸激活的ASICs电流以及下调ASIC3蛋白表达,并对冰醋酸及完全弗氏佐剂诱导的疼痛具有显著抑制效应,且该效应能够被ASICs受体激动剂所抵消,这些均说明TFDB对ASICs的调控可能是其产生镇痛效应的机制之一.