樟树叶挥发性成分研究

通过GC/MS方法,对湖南樟树嫩叶、老叶以及枯叶挥发油的成分和抗氧化性能进行了研究,分析鉴定了其中含量占95%以上的31个化合物.结果表明:嫩叶中以Copaene(28.55%)、石竹烯(25.81%)和α-石竹烯(12.69%)为主要成分;老叶挥发油以芳樟醇含量最高(78.30%);枯叶挥发油主含石竹烯(38.64%...     

土牛膝抗炎成分分离、鉴定与含量测定研究

本研究建立脂多糖诱导的巨噬细胞炎症模型,结合层析法对土牛膝抗炎活性成分进行分离,并用NMR、MS以及与对照品比较,对得到的化合物进行结构鉴定。结果表明粗毛牛膝醇提取物抗炎效果最佳,其反相层析50%、70%及100%甲醇洗脱物抗炎活性高和细胞毒性低,从50%～70%甲醇洗脱物分离鉴定出9种单体化合物:β-蜕皮甾酮(1)、牛膝甾酮(2)、水龙骨甾酮B(3)、N-反式阿魏酰酪胺(4)、N-顺式阿魏酰酪胺(5)、N-顺式阿魏酰-3-甲氧基酪胺(6)、N-反式阿魏酰-3-甲氧基酪胺(7)、竹节参皂苷Ⅳa(8)、5,2′-二甲氧基-6-甲氧甲基-7-羟基-异黄酮(10);100%甲醇洗脱物鉴定出主要成分竹节参皂苷Ⅰ(9)。在LPS+25μM单体浓度下,化合物抗炎活性由强到弱为41526910378。反相液相色谱测定结果表明,野生牛膝和柳叶牛膝(两年生)地下根茎β-蜕皮甾酮含量在温度高季节含量高,8月份达到最高,分别为0.914±0.016和1.412±0.038 mg/g;两年生粗毛牛膝随季节变化规律不明显。具有抗炎活性的阿魏酰酪胺类生物碱首次从土牛膝中分离鉴定。     

不同杂交兰品种花朵挥发性成分分析

采用顶空–固相微萃取技术,分析杂交兰不同品种、不同花期和花器官不同部位的花香成分。结果表明,杂交兰花香的主要成分为萜烯类化合物。不同品种间花香释放量差异明显,‘K24’以桉油精(23.91%)和正己醇(13.74%)为主,‘K21-1’以反式–橙花叔醇(30.39%)和环己烷(22.99%)为主;‘红美人’以α-法呢烯(43.50%)和芳樟醇(34.52%)为主;‘K18’以石竹烯(43.57%)和α-香柑油烯(19.59%)为主,‘黄金龙’以β-月桂烯(25.23%)和α-香柑油烯(15.45%)为主;‘十八格格’以β-石竹烯(42.99%)和α-法呢烯(19.65%)为主;‘汉城公主’以β-石竹烯(52.40%)和α-法呢烯(9.99%)为主。‘K18’释放量和化合物数量在盛开期最高。在花器官不同部位中,花瓣和萼片主要释放β-石竹烯,唇瓣主要释放β-罗勒烯。     

广东紫珠中的萜类化合物及其抗炎活性

对岭南药材广东紫珠(Callicarpa kwangtungensis)地上部分进行化学成分研究,得到11个萜类化合物,分别鉴定为sambucunlin A(1)、2α-羟基羽扇豆醇(2)、swinhoeic acid(3)、3β-羟基-乌苏烷-11-烯-13β,28-内酯(4)、蔷薇酸(5)、2α,3β,6β,18β,23-pentahydroxy-olean-12-en-28-oic acid(6)、rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo-oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid(7)、salvionoside B(8)、齐墩果酸(9)、白桦脂酸(10)和α-香树脂醇(11)。其中,化合物1～4和6～8为首次从该属植物中分离得到。在化学成分分离基础上,进一步选择脂多糖(LPS)诱导的RAW 264.7小鼠巨噬细胞炎症模型进行萜类化合物抗炎活性测试。结果表明:化合物3和9具有显著的抗炎活性,对比结构发现,三萜类化合物(3～6、9和11)抗炎活性优于倍半萜类化合物(7和...     

云南枸杞根中酰胺类化学成分及其抗炎活性研究

研究云南枸杞(Lycium yunnanense Kuang)根部的酰胺类化学成分。通过硅胶和Sephadex LH-20柱层析,以及高效液相制备层析等分离纯化手段,从云南枸杞根部醇提物中共分离得到15个酰胺类化合物,运用现代波谱技术鉴定了它们的结构,分别为反式-N-阿魏酰酪胺(1)、N-顺式-对羟基苯乙基阿魏酰胺(2)、dihydro-feruloyltyramine(3)、3-(4-hydroxy-3-methoxyphenyl)-N-[2-(4-hydroxyphenyl)-2-methoxyethyl]acrylamide(4)、N-反式桂皮酸酰对羟基苯乙胺(5)、反式-N-对羟基香豆酰酪胺(6)、反式-N-对羟基苯乙基咖啡酰胺(7)、二氢咖啡酰酪胺(8)、N-trans-coumaroyloctopamine(9)、N-[2-(3,4-dih-ydroxyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl)prop-2-enamide(10)、N-trans-feruloyl-3'-O-methyldopamine(11)、dihydro-feruloyl-5-methoxytyramine(12)、N-trans-sinapoyltyramine(13)、N-acetyl-N'-trans-feruloylputrescine(14)、N-acetyltyramine(15)。其中,化合物3~5,9~15为首次从该植物中分离得到。对分离得到的15个酰胺类化合物进行抗炎活性筛选,其中,化合物4、6、7和12具有显著的抗炎活性,其IC50值均小于17.21±0.50μM。     

云南枸杞根中酰胺类化学成分及其抗炎活性研究北大核心CSCD

研究云南枸杞(Lycium yunnanense Kuang)根部的酰胺类化学成分。通过硅胶和Sephadex LH-20柱层析,以及高效液相制备层析等分离纯化手段,从云南枸杞根部醇提物中共分离得到15个酰胺类化合物,运用现代波谱技术鉴定了它们的结构,分别为反式-N-阿魏酰酪胺(1)、N-顺式-对羟基苯乙基阿魏酰胺(2)、dihydro-feruloyltyramine(3)、3-(4-hydroxy-3-methoxyphenyl)-N-[2-(4-hydroxyphenyl)-2-methoxyethyl]acrylamide(4)、N-反式桂皮酸酰对羟基苯乙胺(5)、反式-N-对羟基香豆酰酪胺(6)、反式-N-对羟基苯乙基咖啡酰胺(7)、二氢咖啡酰酪胺(8)、N-trans-coumaroyloctopamine(9)、N-[2-(3,4-dih-ydroxyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl)prop-2-enamide(10)、N-trans-feruloyl-3'-O-methyldopamine(11)、dihydro-feruloyl-5-methoxytyramine(12)、N-trans-sinapoyltyramine(13)、N-acetyl-N'-trans-feruloylputrescine(14)、N-acetyltyramine(15)。其中,化合物3~5,9~15为首次从该植物中分离得到。对分离得到的15个酰胺类化合物进行抗炎活性筛选,其中,化合物4、6、7和12具有显著的抗炎活性,其IC50值均小于17.21±0.50μM。     

中国特有植物银杉叶精油成分的研究

本文用气相色谱-质谱-计算机联用仪,分离了银杉(Cathaya argyrophylla)叶精油主要成分26个,鉴定了19个化合物,并测定了含量。其中主要成分有α-蒎烯、β-蒎烯、α-龙脑烯醛、反式-松香芹醇、桃金娘烯醛、马鞭草烯酮和石竹烯氧化物等。     

草八角精油化学成分的研究

采用PGC,GC/MS及IR方法研究了草八角精油化学成分为:芳樟醇(0.08％)、胡椒酚甲醚(21.94％)、顺式大茴香醚(0.03％)、茴香醛(0.05％)、反式大茴香醚(75.39％)、大茴香丙酮(0.03％)、石竹烯(0.08％)、蛇麻烯(0.15％)、异愈疮木烯(0.01％)等10个组分。为进一步开发利用提供依据。     

猫须草抗炎活性成分研究

为了研究猫须草(Clerodendranthus spicatus)抗炎活性成分,该研究采用活性追踪的方法,利用硅胶、MCI柱色谱以及HPLC等分离技术,对猫须草抗炎活性部位进行分离纯化,通过波谱数据分析和文献比对鉴定化合物结构,并利用LPS诱导的RAW 264.7巨噬细胞炎症模型进行抗炎活性评价。结果表明:(1)从猫须草抗炎活性部位分离得到10个化合物,分别鉴定为泡桐素(1)、鼠尾草素(2)、对苯二甲酸二辛酯(3)、N-(N-苯甲酰基-L-苯丙酰基)-L-苯基丙醇(4)、fragransin B₁(5)、6,7,8,4''-四甲氧基黄酮(6)、N-反式-阿魏酰酪胺(7)、N-顺式-阿魏酰酪胺(8)、trans-N-cinnamoyltyramine(9)、新海胆灵 A(10),其中化合物1、4、8-10为首次从该植物中分离得到。(2)抗炎结果显示,猫须草抗炎活性成分主要存在于中低极性部位,从中分离得到的大部分化合物显示出一定的NO生成抑制活性,其中酰胺类成分(7-9)均具有较好的抗炎活性,表明该类成分是猫须草抗炎作用的主要成分之一。该研究丰富了猫须草抗炎物质基础,为其开发利用提供科学依据。     

火麻仁挥发油的化学成分研究

本文采用水蒸气蒸馏法提取传统中药火麻仁中的挥发油,用GC毛细管柱进行分析,面积归一化法测定其相对含量,并用GC-MS鉴定其化学成分.共检出90个色谱峰,鉴定其中62个化合物,占挥发油总量的89 25%.鉴定的成分包括44种烃类化合物、7种芳香族化合物和11种脂肪族化合物及其衍生物.烃类化合物中以萜类为主,占挥发油总量的32.01%,相对含量较高的单萜及其衍生物包括β-芳樟醇(1.66%)、β-顺式-罗勒烯(1.30%)、樟脑(0.93%)、冰片和薄荷醇(0.44%);相对含量较高的倍半萜及其衍生物包括大牦牛儿烯 D(8.20%)、α-芹子烯(5.23%)、β-丁香烯(3.65%)、氧化石竹烯(2.22%)、δ-愈创木烯(1.55%)和α-丁香烯(1.19%).     

The molecular mechanisms of the effect of Dexamethasone and Cyclosporin A on TLR4 /NF-κB signaling pathway activation in oral lichen planus

Toll-like receptors (TLRs) and the nuclear factor-kappa B (NF-κB) signaling transduction pathway play important roles in the pathogenesis of several chronic inflammatory diseases, but its function in oral lichen planus (OLP) remains unclear. In this study, we examined the expression of TLR4 and NF-κB-p65 and inflammatory cytokines TNF-α and IL-1β by immunohistochemistry in OLP tissues, and found that TLR4 and NF-κB-p65 were significantly upregulated in OLP compared to normal oral mucosa (P<0.05). We used keratinocytes HaCaT stimulated with lipopolysaccharide (LPS) to simulate the local OLP immune environment to some extent. RT-PCR and immunoblotting analyses showed significant activation of TLR4 and NF-κB-p65 in the circumstance of LPS-induced inflammatory response. The high expression of TLR4 and NF-κB-p65 are correlated with expression of cytokines TNF-α and IL-1β (P<0.05). We further showed that NF-κB could act as an anti-apoptotic molecule in OLP. We conclude that TLR4 and the NF-κB signaling pathway may interact with the perpetuation of OLP. Steroids and cyclosporine are effective in the treatment of symptomatic OLP. However, there was some weak evidence for the mechanism over Dexamethasone (DeX) and Cyclosporine A (CsA) for the palliation of symptomatic OLP. In the present study, we found that Dexamethasone and Cyclosporine A negatively regulated NF-κB signaling pathway under LPS simulation in HaCaT cells by inhibiting TLR4 expression, on the other hand, Cyclosporine A could inhibit HaCaT cell proliferation by the induction of the apoptosis of HaCaT cells to protect OLP from the destruction of epidermal cells effectively.     

CO-releasing molecules and increased heme oxygenase-1 induce protein S-glutathionylation to modulate NF-κB activity in endothelial cells

Protein glutathionylation is a protective mechanism that functions in response to mild oxidative stress. Carbon monoxide (CO) can increase the reactive oxygen species concentration from a low level via the inhibition of cytochrome c oxidase. We therefore hypothesized that CO would induce NF-κB-p65 glutathionylation and then show anti-inflammatory effects. In this study, we found that CO-releasing molecules suppress TNFα-induced monocyte adhesion to endothelial cells (ECs) and reduce ICAM-1 expression. Moreover, CO donors were further found to exert their inhibitory effects by blocking NF-κB-p65 nuclear translocation, but do so independent of IκBα degradation, in TNFα-treated ECs. In addition, p65 protein glutathionylation represents the response signal to CO donors and is reversed by the reducing agent dithiothreitol. Thiol modification of the cysteine residue in the p65 RHD region was required for the CO-modulated NF-κB activation. The suppression of p65 glutathionylation by a GSH synthesis inhibitor, BSO, and by catalase could also attenuate TNFα-induced p65 nuclear translocation and ICAM-1 expression. CO donors induce Nrf2 activation and Nrf2 siRNA suppresses CO-induced p65 glutathionylation and inhibition. Furthermore, we found that the CO donors induce heme oxygenase-1 (HO-1) expression, which increases p65 glutathionylation. In contrast, HO-1 siRNA attenuates CO donor- and hemin-induced p65 glutathionylation. Our results thus indicate that the glutathionylation of p65 is likely to be responsible for CO-mediated NF-κB inactivation and that the HO-1-dependent pathway may prolong the inhibitory effects of CO donors upon TNFα treatment of ECs.     

The glutathionylation of p65 modulates NF-κB activity in 15-deoxy-Δ¹²,¹⁴-prostaglandin J₂-treated endothelial cells

Protein glutathionylation is a posttranslational modification of cysteine residues with glutathione in response to mild oxidative stress. Because 15-deoxy-Δ12,14-prostaglandin J(2) (15d-PGJ(2)) is an electrophilic prostaglandin that can increase glutathione (GSH) levels and augment reactive oxygen species (ROS) production, we hypothesized that it induces NF-κB-p65 glutathionylation and would exert anti-inflammatory effects. Herein, we show that 15d-PGJ(2) suppresses the expression of ICAM-1 and NF-κB-p65 nuclear translocation. 15d-PGJ(2) upregulates the Nrf2-related glutathione synthase gene and thereby increases the GSH levels. Consistent with this, Nrf2 siRNA molecules abolish the inhibition of p65 nuclear translocation in 15d-PGJ(2)-induced endothelial cells (ECs). ECs treated with GSSG show increased thiol modifications of p65 and also a block in TNFα-induced p65 nuclear translocation and ICAM-1 expression, but not in IκBα degradation. However, the overexpression of glutaredoxin 1 was found to be accompanied by a modest increase in NF-κB activity. Furthermore, we found that multiple cysteine residues in p65 are responsible for glutathionylation. 15d-PGJ(2) was observed to induce p65 glutathionylation and is suppressed by a GSH synthesis inhibitor, buthionine sulfoximine, by catalase, and by Nrf2 siRNA molecules. Our results thus indicate that the GSH/ROS-dependent glutathionylation of p65 is likely to be responsible for 15d-PGJ(2)-mediated NF-κB inactivation and for the enhanced inhibitory effects of 15d-PGJ(2) on TNFα-treated ECs.     

Proepithelin stimulates growth plate chondrogenesis via nuclear factor-kappaB-p65-dependent mechanisms

Proepithelin, a previously unrecognized growth factor in cartilage, has recently emerged as an important regulator for cartilage formation and function. In the present study, we provide several lines of evidences in proepithelin-mediated induction of cell proliferation, differentiation, and apoptosis in the metatarsal growth plate. Proepithelin-mediated stimulation of metatarsal growth and growth plate chondrogenesis was neutralized by pyrrolidine dithiocarbamate, a known NF-κB inhibitor. In rat growth plate chondrocytes, proepithelin induced NF-κB-p65 nuclear translocation, and nuclear NF-κB-p65 initiated its target gene cyclin D1 to regulate chondrocyte functions. The inhibition of NF-κB-p65 expression and activity (by p65 short interfering RNA (siRNA) and pyrrolidine dithiocarbamate, respectively) in chondrocytes reversed the proepithelin-mediated induction of cell proliferation and differentiation and the proepithelin-mediated prevention of cell apoptosis. Moreover, the inhibition of the phosphatidylinositol 3-kinase and Akt abolished the effects of proepithelin on NF-κB activation. Finally, using siRNA and antisense strategies, we demonstrated that endogenously produced proepithelin by chondrocytes is important for chondrocyte growth in serum-deprived conditions. These results support the hypothesis that the induction of NF-κB activity of in growth plate chondrocytes is critical in proepithelin-mediated growth plate chondrogenesis and longitudinal bone growth.     

Honokiol: an effective inhibitor of tumor necrosis factor-α-induced up-regulation of inflammatory cytokine and chemokine production in human synovial fibroblasts

In this study, we investigated the mechanisms underlying the anti-inflammatory effects of honokiol in tumor necrosis factor (TNF)-α-stimulated rheumatoid arthritis synovial fibroblasts (RASFs). RASFs pre-treated with honokiol (0-20 μM) were stimulated with TNF-α (20 ng/ml). The levels of prostaglandin E2 (PGE2), nitric oxide (NO), soluble intercellular adhesion molecule-1 (sICAM-1), transforming growth factor-β1 (TGF-β1), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) in supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and Griess assay. In addition, protein expression levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and phosphorylated Akt, nuclear factor kappa B (NFκB), and extracellular signal-regulated kinase (ERK)1/2 were determined by western blot. The expression of NFκB-p65 was assessed by immunocytochemical analysis. TNF-α treatment significantly up-regulated the levels of PGE2, NO, sICAM-1, TGF-β1, MCP-1, and MIP-1α in the supernatants of RASFs, increased the protein expression of COX-2, iNOS, and induced phosphorylation of Akt, IκB-α, NFκB, and ERK1/2 in RASFs. TNF-α-induced expression of these molecules was inhibited in a dose-dependent manner by pre-treatment with honokiol. The inhibitory effect of honokiol on NFκB-p65 activity was also confirmed by immunocytochemical analysis. In conclusion, honokiol is a potential inhibitor of TNF-α-induced expression of inflammatory factors in RASFs, which holds promise as a potential anti-inflammatory drug.     

The spin trap 5,5-dimethyl-1-pyrroline N-oxide inhibits lipopolysaccharide-induced inflammatory response in RAW 264.7 cells

AimExposure of macrophages to lipopolysaccharide (LPS) induces oxidative and inflammatory stresses, which cause cell damage. Antioxidant and anti-inflammatory properties have been attributed to the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO), commonly used in free radical analysis, but these aspects of DMPO have been little explored. In this study, we sought to establish the anti-inflammatory activity of DMPO, presumably by removing free radicals which otherwise help activate inflammatory response and damage cells.Main methodsRAW 264.7 macrophages were treated with LPS and/or DMPO for different time points, cell damage, production of inflammatory mediators, inducible nitric oxide synthase (iNOS) expression, NF-κB p65 activation, phosphorylation of MAPKs and Akt, and intracellular reactive oxygen species (ROS) were determined.Key findingsAfter cells were treated with LPS and/or DMPO for 24 h, DMPO reduced the LPS-induced inflammatory response as indicated by downregulated iNOS expression and production of inflammatory mediators. Accordingly, DMPO protected cells from LPS-induced cytotoxicity. In order to understand the mechanistic basis of these DMPO effects, the NF-κB p65 activation and the phosphorylation of MAPKs and Akt were examined. We found, by assaying cells treated with LPS and/or DMPO for 15–60 min, that DMPO inhibited the phosphorylation of MAPKs, Akt, and IκBα, and reduced the NF-κB p65 translocation. Furthermore, we demonstrated that DMPO inhibited LPS-induced ROS production.SignificanceDMPO showed the anti-inflammatory activity and attenuated LPS-induced cell damage, most likely by reducing ROS production and thus preventing the subsequent inflammatory activation and damage.