可溶性环氧化物水解酶抑制剂对小鼠内皮祖细胞分泌血管内皮生长因子及缺氧诱导因子-1α的促进作用

目的 探讨可溶性环氧化物水解酶抑制剂(t-AUCB)能否促进小鼠来源内皮祖细胞(EPCs)分泌内皮生长因子(VEGF)和缺氧诱导因子-1α(HIF-1α),及可能的相关机制.方法 提取小鼠长骨骨髓,离心分离培养,取得高纯度EPCs,不同浓度可溶性环氧化物水解酶抑制剂(t-AUCB)和过氧化体增殖物激活型受体γ(PPAR...     

丹参水溶性有效成分对内皮祖细胞功能的影响

丹参水溶性有效成分具有广泛的生理活性与药理作用。内皮祖细胞(endothelial progenitor cells,EPCs)是一类具有高增殖潜能并能分化为血管内皮细胞,但尚未表达成熟内皮细胞表型,也未形成血管的前体细胞。研究发现丹参水溶性有效成分可通过调节EPCs功能来修复内皮损伤,从而在内皮损伤相关性疾病中发挥保护及治疗作用。该文综述了丹参水溶性有效成分对EPCs功能的影响,从而为丹参水溶性有效成分的进一步开发利用提供依据。     

辛伐他汀和罗格列酮对内皮祖细胞的影响

目的 探讨辛伐他汀和罗格列酮对内皮祖细胞(EPC)增殖、分化、迁移和集落形成能力的影响.方法 以密度梯度离心法获取人外周血单核细胞,将其接种于人纤连蛋白包被的培养板上,以EBM-2培养基培养.分别采用MTT比色法、改良Boyden小室法、甲基纤维素培养基培养法观察辛伐他汀和罗格列酮对EPC增殖、迁移和集落形成能力的影响...     

当归补血汤含药血清对内皮祖细胞功能及其PI3K/Akt通路影响的研究

目的 研究当归补血汤含药血清对人内皮祖细胞(EPCs)功能及其PI3K/Akt通路的影响.方法 体外培养并鉴定人外周血EPCs,制备当归补血汤含药血清,对EPCs进行不同血清或PI3K抑制剂的干预,检测EPCs的增殖、黏附、迁移、成小管功能及NO分泌量,RT-PCR检测eNOS mRNA的表达,Western blot检测Akt蛋白的表达.结果 与对照组比较,当归补血汤含药血清能促进EPCs的增殖、黏附、迁移及成小管功能(P＜0.05),促进EPCs分泌NO,上调细胞eNOS mRNA、总Akt和磷酸化Akt蛋白的表达,但这些作用均在PI3K抑制剂处理后明显减弱(P＜0.05).结论 当归补血汤可能通过PI3K/Akt通路调节EPCs的功能.     

骨髓动员对大鼠骨髓源内皮祖细胞功能的影响

目的观察大鼠骨髓动员后骨髓源内皮祖细胞(endothelial progenitor cells,EPCs)功能的改变,探讨骨髓动员后骨髓干细胞移植提高下肢缺血性疾病疗效的可能机制。方法按是否接受重组人粒细胞集落刺激因子(recombinant human granulocyte-macrophage colony-stimulatory factor,rhG-CSF)注射液注射将Lewis大鼠分为动员组和对照组,获取骨髓单个核细胞用EBM-2培养液进行诱导分化,培养7d后对贴壁细胞进行EPCs的鉴定;采用黏附能力测定实验观察EPCs的黏附能力。另取Lewis大鼠制备单侧后肢缺血模型,在模型制备后7d将8×106个EPCs移植至缺血侧后肢的肌肉内,按照移植EPCs来源不同将后肢缺血大鼠分为动员组和对照组。EPCs移植4周后行后肢血管造影,计算缺血侧后肢侧支血管数目。结果大鼠骨髓单个核细胞培养7d后黏附能力测定实验中动员组黏附EPCs数量为(22.3±2.8)个/×100高倍视野大于对照组(16.8±4.4)个/×100高倍视野(P<0.05);大鼠骨髓源EPCs移植至缺血肢体后4周动员组EPCs移植后缺血侧后肢侧支血管数目为(4.7±1.0)大于对照组(3.3±0.5)(P<0.05)。结论骨髓动员使骨髓源EPCs功能改善,其可能是骨髓动员后骨髓干细胞移植提高下肢缺血性疾病疗效的机制之一。     

阿托伐他汀对外周血培养内皮祖细胞数量及功能的影响

目的 观察阿托伐他汀对体外培养外周血内皮祖细胞(EPCs)数量及功能的影响.方法 分离培养人外周血单个核细胞,分别接种在普通培养基(A组)及阿托伐他汀干预培养基(B组),7d后收集贴壁细胞进行分析.激光共聚焦显微镜下双染色阳性细胞鉴定为正在分化的EPCs,采用transwell小室、细胞计数法评估EPCs扩增、黏附及迁移能力的差别.结果 与A组相比,B组体外培养EPCs的数量显著增多,黏附及迁移能力显著提高.结论 阿托伐他汀在外周血EPCs培养中能促使细胞扩增,增强黏附及迁移功能,可作为EPCs培养的一种改良方法.     

天然糖水解酶抑制剂

本专利涉及一种活性比传统已知的糖水解酶抑制剂更强的糖水解酶抑制剂，而且是从天然产品中得到的。这种糖水解酶抑制剂组成是Ascophyllum nodsum的提取物，其活性成分是一种棕藻。这种糖水解酶抑制剂以保健食品或特殊的保健食品供应，用来治疗和预防糖尿病。     

改善糖尿病内皮祖细胞功能的研究进展

内皮祖细胞（EPC）为来源于骨髓及外周血中的干细胞,能定向增殖分化为内皮细胞,参与血管损伤后的修复。研究显示,改善糖尿病EPC功能,可以预防和降低糖尿病并发症的发生。追踪近年来改善EPC功能的研究进展,就其对糖尿病并发症的预防和改善作用做一综述,以期为糖尿病的治疗提供新思路。     

诺帝对血管内皮生长因子诱导的人脐血源性内皮祖细胞功能的影响

探讨手性化合物诺帝(Nordy)对血管内皮生长因子(VEGF)诱导的人脐血源性内皮祖细胞(EPCs)功能的影响及其意义。应用密度梯度离心法分离新鲜人脐血的单个核细胞，接种于EGM-2培养液中培养7～10 d获得内皮祖细胞(EPCs)。分别采用MTT法、Millicell-PCF培养小室系统和Matrigel内小管形成试验检测诺帝对VEGF刺激下EPCs增殖活性、迁移能力和体外形成小管样结构能力的影响。结果表明，100 μmol·L^-1诺帝作用24 h明显抑制EPCs增殖活性(P<0.05)，诺帝(25～50 μmol·L^-1)作用48～72 h也明显抑制EPCs增殖活性(P<0.05)。诺帝(25～100 μmol·L^-1)显著抑制VEGF诱导的EPCs迁移活性和体外形成小管样结构的能力(P<0.05)。诺帝能抑制体外VEGF诱导的人脐血源性EPCs增殖、迁移和体外小管形成能力，提示其具有抗EPCs效应。     

Discovery of phthalimide derivatives as novel inhibitors of a soluble epoxide hydrolase

Soluble epoxide hydrolase (sEH) inhibitors are effective in reducing blood pressure, inflammation, and pain in a number of mammalian disease models. As most classical urea-based sEH inhibitors suffer from poor solubility and pharmacokinetic properties, the development of novel sEH inhibitors with an improved pharmacokinetic specification has received a great deal of attention. In this study, a series of amide-based sEH inhibitors bearing a phthalimide ring as the novel secondary pharmacophore (P₂) was designed, synthesized, and evaluated. Docking results illustrated that the amide group as the primary pharmacophore (P₁) was placed at a suitable distance from the three key amino acids (Tyr383, Tyr466, and Asp335) for an effective hydrogen bonding. In agreement with these findings, most of the newly synthesized compounds demonstrated moderate to high sEH inhibitory activities, relative to 12-(3-adamantan-1-yl-ureido)dodecanoic acid as the reference standard. Compound 12e with a 4-methoxybenzoyl substituent exhibited the highest sEH inhibitory activity, with an IC₅₀ value of 1.06 nM. Moreover, the ADME properties of the compounds were evaluated in silico, and the results revealed appropriate predictions.     

Soluble epoxide hydrolase inhibitors: a patent review

Importance of the field: Soluble epoxide hydrolase (sEH) inhibitors have been shown to effectively increase the levels of epoxyeicosatrienoic acids and reduce the levels of dihydroxyeicosatrienoic acids, which may be translated to therapeutic potentials for multiple disease indications. It has been claimed that sEH inhibitors can be used to treat hypertension, diabetes, stroke, dyslipidemia, pain, immunological disorders, eye diseases, neurological diseases and other indications.

Areas covered in this review: A comprehensive synopsis of patent literature on sEH inhibitors is provided.

What the reader will gain: A total of more than 100 patent publications describing multiple classes of sEH inhibitors are analyzed. These include amides, ureas, thioamides, thioureas, carbamates, acyl hydrazones, chalcone oxdies, etc. In addition to selected in vitro and in vivo data of representative sEH inhibitors, a wide range of proposed applications of sEH inhibitors are also summarized.

Take home message: Several sEH inhibitors with potent in vitro and in vivo target inhibition appear promising, including one Phase II clinical candidate. The clinical evaluation will be critical to assess the proclaimed therapeutic utility of sEH inhibition.     

Pharmacokinetics and in vivo potency of soluble epoxide hydrolase inhibitors in cynomolgus monkeys

BACKGROUND AND PURPOSE

Soluble epoxide hydrolase inhibitors (sEHIs) possess anti-inflammatory, antiatherosclerotic, antihypertensive and analgesic properties. The pharmacokinetics (PK) and pharmacodynamics in terms of inhibitory potency of sEHIs were assessed in non-human primates (NHPs). Development of a sEHI for use in NHPs will facilitate investigations on the role of sEH in numerous chronic inflammatory conditions.

EXPERIMENTAL APPROACH

PK parameters of 11 sEHIs in cynomolgus monkeys were determined after oral dosing with 0.3 mg·kg⁻¹. Their physical properties and inhibitory potency in hepatic cytosol of cynomolgus monkeys were examined. Dose-dependent effects of the two inhibitors 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU) and the related acetyl piperidine derivative, 1-trifluoromethoxyphenyl-3-(1-acetylpiperidin-4-yl) urea (TPAU), on natural blood eicosanoids, were determined.

KEY RESULTS

Among the inhibitors tested, TPPU and two 4-(cyclohexyloxy) benzoic acid urea sEHIs displayed high plasma concentrations (>10 × IC₅₀), when dosed orally at 0.3 mg·kg⁻¹. Although the 4-(cyclohexyloxy) benzoic acid ureas were more potent against monkey sEH than piperidyl ureas (TPAU and TPPU), the latter compounds showed higher plasma concentrations and more drug-like properties. The C_max increased with dose from 0.3 to 3 mg·kg⁻¹ for TPPU and from 0.1 to 3 mg·kg⁻¹ for TPAU, although it was not linear over this range of doses. As an indication of target engagement, ratios of linoleate epoxides to diols increased with TPPU administration.

CONCLUSION AND IMPLICATIONS

Our data indicate that TPPU is suitable for investigating sEH biology and the role of epoxide-containing lipids in modulating inflammatory diseases in NHPs.     

Computational insights into the known inhibitors of human soluble epoxide hydrolase

Human soluble epoxide hydrolase (hsEH) is involved in the hydrolysis of epoxyeicosatrienoic acids (EETs), which have potent anti-inflammatory properties. Given that EET conversion generates nonbioactive molecules, inhibition of this enzyme would be beneficial. Past decades of work on hsEH inhibitors resulted in numerous potential compounds, of which a hundred hsEH–ligand complexes were crystallized and deposited in the Protein Data Bank (PDB). We analyzed all deposited hsEH–ligand complexes to gain insight into the binding of inhibitors and to provide feedback on the future drug design processes. We also reviewed computationally driven strategies that were used to propose novel hsEH inhibitors.     

Discovery of soluble epoxide hydrolase inhibitors from natural products

With the goal of developing soluble epoxide hydrolase (sEH) inhibitors with novel chemical structures, the sEH inhibitory activities of 30 natural compounds were evaluated using both a fluorescent substrate, 3-phenyl-cyano(6-methoxy-2-naphthalenyl)methyl ester- 2-oxiraneacetic acid, and a physiological substrate, 14,15-epoxyeicosatrienoic acid. To evaluate the selectivity of sEH inhibition, the inhibition of microsomal epoxide hydrolase (mEH), which plays a critical role in detoxification of toxic epoxides, was determined using human liver microsomes. Honokiol and β-amyrin acetate, isolated from Magnolia officinalis and Acer mandshuricum, respectively, displayed strong inhibition of sEH activity, with respective IC₅₀ values of 0.57 μM and 3.4 μM determined using the fluorescent substrate, and 1.7 μM and 6.1 μM determined using 14,15-epoxyeicosatrienoic acid. mEH activity was decreased to 49% or 61% of control activity by 25 μM honokiol or β-amyrin acetate, respectively. These results suggest that β-amyrin acetate and honokiol exhibit sEH inhibitory activity, although their sEH selectivity should be improved.     

Ginsenoside Rgl promotes endothelial progenitor cell migration and proliferation

Aim： To investigate the effect of ginsenoside Rgl on the migration, adhesion, proliferation, and VEGF expression of endothelial progenitor cells （EPCs）.
Methods： EPCs were isolated from human peripheral blood and incubated with different concentrations of ginsenoside Rgl （0.1, 0.5, 1.0, and 5.0 μmol/L） and vehicle controls. EPC migration was detected with a modified Boyden chamber assay. EPC adhesion was determined by counting adherent cells on fibronectin-coated culture dishes. EPC proliferation was analyzed with the 3.（4,5.dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay. In vitro vasculogenesis was assayed using an in vitro vasculogenesis detection kit. A VEGF-ELISA kit was used to measure the amount of VEGF protein in the cell culture medium.
Results： Ginsenoside Rgl promoted EPC adhesion, proliferation, migration and in vitro vasculogenesis in a dose- and timedependent manner. Cell cycle analysis showed that 5.0 μmol/L ofginsenoside Rgl significantly increased the EPC proliferative phase （S phase） and decreased the resting phase （G0/G1 phase）. Ginsenoside Rgl increased vascular endothelial growth factor production. Conclusion： The results indicate that ginsenoside Rgl promotes proliferation, migration, adhesion and in vitro vasculogenesis.     

Ginsenoside Rg1 promotes endothelial progenitor cell migration and proliferation

Aim:

To investigate the effect of ginsenoside Rg1 on the migration, adhesion, proliferation, and VEGF expression of endothelial progenitor cells (EPCs).

Methods:

EPCs were isolated from human peripheral blood and incubated with different concentrations of ginsenoside Rg1 (0.1, 0.5, 1.0, and 5.0 μmol/L) and vehicle controls. EPC migration was detected with a modified Boyden chamber assay. EPC adhesion was determined by counting adherent cells on fibronectin-coated culture dishes. EPC proliferation was analyzed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. In vitro vasculogenesis was assayed using an in vitro vasculogenesis detection kit. A VEGF-ELISA kit was used to measure the amount of VEGF protein in the cell culture medium.

Results:

Ginsenoside Rg1 promoted EPC adhesion, proliferation, migration and in vitro vasculogenesis in a dose- and time-dependent manner. Cell cycle analysis showed that 5.0 μmol/L of ginsenoside Rg1 significantly increased the EPC proliferative phase (S phase) and decreased the resting phase (G₀/G₁ phase). Ginsenoside Rg1 increased vascular endothelial growth factor production.

Conclusion:

The results indicate that ginsenoside Rg1 promotes proliferation, migration, adhesion and in vitro vasculogenesis.