The inhibitory effect of astilbin on transplant arteriosclerosis in murine model of thoracic aorta transplantation was examined. Model of rat thoracic aorta transplantation was established. Ninety rats were divided in...The inhibitory effect of astilbin on transplant arteriosclerosis in murine model of thoracic aorta transplantation was examined. Model of rat thoracic aorta transplantation was established. Ninety rats were divided into three groups. In isograft group, the thoracic aorta of Brown Norway （BN） rat was anastomosed with the abdominal aorta of another BN rat. In allograft group, the thoracic aorta of BN rat was anastomosed with the abdominal aorta of Lewis rat. In astilbin group, the rats receiving allo-transplantation were given astilbin 5 mg/kg per day for a time of 28 days. The donor thoracic aorta and the recipient abdominal aorta were anastomosed by means of a polyethylene can- nula （inner diameter： 1.5 mm, length： 3 mm length）. The grafts were histologically examined for structural changes. The areas of arterial lumen and endatrium were calculated. Our results showed that, in the allograft group, 28 days after allografting, conspicuous proliferation of smooth muscles and infiltration with a great number of inflammatory cells were found in the tunica intima and tunica media. Astilbin significantly inhibited the proliferation of smooth muscles and ameliorated the infiltration of inflammatory cells thereyby prevent against the development of transplant arteriosclerosis. It is concluded that asltilbin can effectively prevent the development of arteriosclerosis in allotransplant by inhibiting the proliferation of smooth muscles and inhibit the proliferation of smooth muscles in tunica of intima and media and reducing infiltration of the inflammatory cells.展开更多
This study examined the effect of astilbin on the proliferation of rat aortic smooth muscle cells (RASMCs) induced by angiotensin Ⅱ (AngⅡ) and explored the possible mechanisms. Cell proliferation model of RASMCs was...This study examined the effect of astilbin on the proliferation of rat aortic smooth muscle cells (RASMCs) induced by angiotensin Ⅱ (AngⅡ) and explored the possible mechanisms. Cell proliferation model of RASMCs was induced by treatmente with AngⅡ. Cells were randomly divided to 8 groups. Normally cultured VSMCs serves as blank control group; in AngⅡ model group, cells were treated with AngⅡ at 10–7 mol/L; in three astilbin groups, cells were treated with 10, 15, 30 mg/L of astilbin; in three AngⅡ+astilbin groups, cells were treated with AngⅡ (at 10–7 mol/L) and astilbin at 10, 15, 30 mg/L. Cell proliferation ability was detected by MTT method and the cell cycles and proliferation index were flow cytometrically determined. The expression of c-myc mRNA was assessed by using reverse transcription polymerase chain reaction (RT-PCR), and the expression of NF-κB in RASMCs was immunocytochemically observed. Our results showed that MTT metabo-lism in RASMCs in the basic and AngII stimulated situation was inhibited by astilbin, and the cells numbers of G0/G1 phase were increased and that of G2/S phase were decreased markedly. Not only highly expression of c-myc gene stimulated by AngⅡ could be inhibited by Astilbin significantly, but also the expression of NF-κB protein can be down regulated by Astilbin. We are led to conclude that astilbin astilbin can inhibit the AngⅡ-mediated proliferation of RASMCs by blocking the transition of RASMCs from G0/G1 phase to S phase and by down-regulating the expression of NF-κB, c-myc gene.展开更多
Objective:To investigate the osteoblastogenic activity of the ethyl acetate(EtOAc)extract of Smilax glabra Roxb roots and its major active compound astilbin.Methods:Astilbin was isolated from EtOAc extract using silic...Objective:To investigate the osteoblastogenic activity of the ethyl acetate(EtOAc)extract of Smilax glabra Roxb roots and its major active compound astilbin.Methods:Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization.Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature.MTT method was used to detect the toxicity.Alkaline phosphatase(ALP)activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate.Calcium deposition was stained with alizarin red-S,distained with cetylpyridium chloride,and quantified at 562 nm.In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6.The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR.Results:Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0,10,25,and 50μg/mL.At 25μg/mL,they enhanced ALP activity and mineralization of osteoblasts up to 30%and 55%for the EtOAc extract and 22%and 41%for astilbin,respectively.Molecular docking analysis of astilbin-ALP interaction revealed Arg167,Asp320,His324,and His437 were key residues participating in hydrophobic interaction;meanwhile,His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP.Moreover,the expression level of genes opn,col1,osx,and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2;3.7;4.1;2.3,respectively at 10μg/mL(P<0.05).Conclusions:The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation.It could be a new and promising osteogenic agent with dual actions for therapeutic applications.展开更多
Astilbin is a potential immunosuppressive agent with minor cytotoxicity. Its oral bioavailability is supposed to be rather low and therefore a sensitive analytical method is required for its pharmacokinetic study afte...Astilbin is a potential immunosuppressive agent with minor cytotoxicity. Its oral bioavailability is supposed to be rather low and therefore a sensitive analytical method is required for its pharmacokinetic study after oral administration. A simple, sensitive and rapid liquid chromatography-tandem mass spectrometry（LC-MS/MS） method was developed and validated for the determination of astilbin in rat plasma. Plasma samples were subjected to liquid-liquid extraction with ethyl acetate and separated by reversed phase high performance liquid chromatography（HPLC） with methanol-0.01%（volume fraction） formic acid（50：50, volume ratio） as mobile phase. Quantitive determination was achieved on negative LC-MS/MS by a multiple reaction moitoring method with transitions m/z 449.1→150.9（quantifier） and m/z 449.1→284.9（qualifier） for astilbin and m/z 128.9→42.0 for internal standard（IS）. A lower limit of quantification（LLOQ） of ng/mL was achieved within a short cycle time of 3.4 min. The method was successfully applied to a pharmacokinetic study involving oral and intravenous administrations of 6 mg/kg astilbin to six rats.展开更多
为建立火索藤(Bauhinia aurea)的鉴别方法及含量测定方法,解决其真伪辨别的问题,分析其药效不一的原因,该文采用基原鉴别、性状鉴别、显微鉴别、薄层色谱法及高效液相色谱法对火索藤开展生药学研究。结果表明:(1)火索藤为多年生粗壮木...为建立火索藤(Bauhinia aurea)的鉴别方法及含量测定方法,解决其真伪辨别的问题,分析其药效不一的原因,该文采用基原鉴别、性状鉴别、显微鉴别、薄层色谱法及高效液相色谱法对火索藤开展生药学研究。结果表明:(1)火索藤为多年生粗壮木质藤本,其茎、叶、花、果实表面均被红棕色茸毛。(2)茎横切面皮层可见大量石细胞群及晶纤维,髓部常见草酸钙簇晶、方晶;叶横切面显示其叶为异面叶,皮层可见草酸钙簇晶、方晶,韧皮部可见分泌腔断续排列成环,韧皮部外侧常见晶纤维。粉末中可见草酸钙簇晶、晶纤维、石细胞、具缘纹孔导管、非腺毛、气孔。(3)各批次火索藤TLC斑点与对照品金丝桃苷、落新妇苷、槲皮苷在薄层色谱图的相同位置上显相同颜色荧光。(4)落新妇苷和槲皮苷进样量分别在0.005888~2.355μg(r=0.9996)、0.03955~1.582μg(r=0.9998)与峰面积呈现良好的线性关系,加样回收率分别为96.42%、104.20%,相对标准偏差(relative standard deviation,RSD)值分别为2.55%、1.79%。该文建立了火索藤的生药学鉴别方法和一种同时测定其中落新妇苷、槲皮苷的含量测定方法,该方法简便、稳定、准确,可为该药材质量标准的制定提供依据。展开更多
目的:采用反相高效液相色谱法,测定不同来源土茯苓药材中落新妇苷和白藜芦醇的含量。方法:采用 YMC-PackODS-A(5 μm,4.6 mm×250 mm)色谱柱,乙腈(A)-0.05%磷酸水溶液(B)梯度洗脱(0～10 min 时25%A,10～15 min 时25%→35%A,15～20 m...目的:采用反相高效液相色谱法,测定不同来源土茯苓药材中落新妇苷和白藜芦醇的含量。方法:采用 YMC-PackODS-A(5 μm,4.6 mm×250 mm)色谱柱,乙腈(A)-0.05%磷酸水溶液(B)梯度洗脱(0～10 min 时25%A,10～15 min 时25%→35%A,15～20 min 时35%A);流速0.8 mL·min^(-1);检测波长306 nm。结果:落新妇苷、白藜芦醇浓度分别在0.018～0.288 mg·mL^(-1)和0.0025～0.025 mg·mL^(-1)范围内,与峰面积呈良好线性关系。落新妇苷平均回收率为99.6%～102.1%,RSD<1.9%;白藜芦醇平均回收率为99.2%～101.8%,RSD<2.2%。结论:该方法简便、准确,重复性好,可作为土茯苓药材的质量控制方法。展开更多
基金supported by a grant from the National Natural Sciences Foundation of China (No.30500656)
文摘The inhibitory effect of astilbin on transplant arteriosclerosis in murine model of thoracic aorta transplantation was examined. Model of rat thoracic aorta transplantation was established. Ninety rats were divided into three groups. In isograft group, the thoracic aorta of Brown Norway （BN） rat was anastomosed with the abdominal aorta of another BN rat. In allograft group, the thoracic aorta of BN rat was anastomosed with the abdominal aorta of Lewis rat. In astilbin group, the rats receiving allo-transplantation were given astilbin 5 mg/kg per day for a time of 28 days. The donor thoracic aorta and the recipient abdominal aorta were anastomosed by means of a polyethylene can- nula （inner diameter： 1.5 mm, length： 3 mm length）. The grafts were histologically examined for structural changes. The areas of arterial lumen and endatrium were calculated. Our results showed that, in the allograft group, 28 days after allografting, conspicuous proliferation of smooth muscles and infiltration with a great number of inflammatory cells were found in the tunica intima and tunica media. Astilbin significantly inhibited the proliferation of smooth muscles and ameliorated the infiltration of inflammatory cells thereyby prevent against the development of transplant arteriosclerosis. It is concluded that asltilbin can effectively prevent the development of arteriosclerosis in allotransplant by inhibiting the proliferation of smooth muscles and inhibit the proliferation of smooth muscles in tunica of intima and media and reducing infiltration of the inflammatory cells.
基金supported by agrant from the National Natural Science Foundation of China(No.30500656)
文摘This study examined the effect of astilbin on the proliferation of rat aortic smooth muscle cells (RASMCs) induced by angiotensin Ⅱ (AngⅡ) and explored the possible mechanisms. Cell proliferation model of RASMCs was induced by treatmente with AngⅡ. Cells were randomly divided to 8 groups. Normally cultured VSMCs serves as blank control group; in AngⅡ model group, cells were treated with AngⅡ at 10–7 mol/L; in three astilbin groups, cells were treated with 10, 15, 30 mg/L of astilbin; in three AngⅡ+astilbin groups, cells were treated with AngⅡ (at 10–7 mol/L) and astilbin at 10, 15, 30 mg/L. Cell proliferation ability was detected by MTT method and the cell cycles and proliferation index were flow cytometrically determined. The expression of c-myc mRNA was assessed by using reverse transcription polymerase chain reaction (RT-PCR), and the expression of NF-κB in RASMCs was immunocytochemically observed. Our results showed that MTT metabo-lism in RASMCs in the basic and AngII stimulated situation was inhibited by astilbin, and the cells numbers of G0/G1 phase were increased and that of G2/S phase were decreased markedly. Not only highly expression of c-myc gene stimulated by AngⅡ could be inhibited by Astilbin significantly, but also the expression of NF-κB protein can be down regulated by Astilbin. We are led to conclude that astilbin astilbin can inhibit the AngⅡ-mediated proliferation of RASMCs by blocking the transition of RASMCs from G0/G1 phase to S phase and by down-regulating the expression of NF-κB, c-myc gene.
基金supported by the the Vietnam Academy of Science and Technology under grant NCCC 08.10/20-20the Institute of Biotechnology under grant CS20-01。
文摘Objective:To investigate the osteoblastogenic activity of the ethyl acetate(EtOAc)extract of Smilax glabra Roxb roots and its major active compound astilbin.Methods:Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization.Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature.MTT method was used to detect the toxicity.Alkaline phosphatase(ALP)activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate.Calcium deposition was stained with alizarin red-S,distained with cetylpyridium chloride,and quantified at 562 nm.In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6.The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR.Results:Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0,10,25,and 50μg/mL.At 25μg/mL,they enhanced ALP activity and mineralization of osteoblasts up to 30%and 55%for the EtOAc extract and 22%and 41%for astilbin,respectively.Molecular docking analysis of astilbin-ALP interaction revealed Arg167,Asp320,His324,and His437 were key residues participating in hydrophobic interaction;meanwhile,His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP.Moreover,the expression level of genes opn,col1,osx,and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2;3.7;4.1;2.3,respectively at 10μg/mL(P<0.05).Conclusions:The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation.It could be a new and promising osteogenic agent with dual actions for therapeutic applications.
基金the National Natural Science Foundation of China，the Science and Technology Major Specialized Projects for ＂Significant New Drugs Creation＂ of the 12th Five-year Plan of China，the National Key Technology R＆D Program of the Ministry of Science and Technology,China
文摘Astilbin is a potential immunosuppressive agent with minor cytotoxicity. Its oral bioavailability is supposed to be rather low and therefore a sensitive analytical method is required for its pharmacokinetic study after oral administration. A simple, sensitive and rapid liquid chromatography-tandem mass spectrometry（LC-MS/MS） method was developed and validated for the determination of astilbin in rat plasma. Plasma samples were subjected to liquid-liquid extraction with ethyl acetate and separated by reversed phase high performance liquid chromatography（HPLC） with methanol-0.01%（volume fraction） formic acid（50：50, volume ratio） as mobile phase. Quantitive determination was achieved on negative LC-MS/MS by a multiple reaction moitoring method with transitions m/z 449.1→150.9（quantifier） and m/z 449.1→284.9（qualifier） for astilbin and m/z 128.9→42.0 for internal standard（IS）. A lower limit of quantification（LLOQ） of ng/mL was achieved within a short cycle time of 3.4 min. The method was successfully applied to a pharmacokinetic study involving oral and intravenous administrations of 6 mg/kg astilbin to six rats.
文摘为建立火索藤(Bauhinia aurea)的鉴别方法及含量测定方法,解决其真伪辨别的问题,分析其药效不一的原因,该文采用基原鉴别、性状鉴别、显微鉴别、薄层色谱法及高效液相色谱法对火索藤开展生药学研究。结果表明:(1)火索藤为多年生粗壮木质藤本,其茎、叶、花、果实表面均被红棕色茸毛。(2)茎横切面皮层可见大量石细胞群及晶纤维,髓部常见草酸钙簇晶、方晶;叶横切面显示其叶为异面叶,皮层可见草酸钙簇晶、方晶,韧皮部可见分泌腔断续排列成环,韧皮部外侧常见晶纤维。粉末中可见草酸钙簇晶、晶纤维、石细胞、具缘纹孔导管、非腺毛、气孔。(3)各批次火索藤TLC斑点与对照品金丝桃苷、落新妇苷、槲皮苷在薄层色谱图的相同位置上显相同颜色荧光。(4)落新妇苷和槲皮苷进样量分别在0.005888~2.355μg(r=0.9996)、0.03955~1.582μg(r=0.9998)与峰面积呈现良好的线性关系,加样回收率分别为96.42%、104.20%,相对标准偏差(relative standard deviation,RSD)值分别为2.55%、1.79%。该文建立了火索藤的生药学鉴别方法和一种同时测定其中落新妇苷、槲皮苷的含量测定方法,该方法简便、稳定、准确,可为该药材质量标准的制定提供依据。
文摘目的:采用反相高效液相色谱法,测定不同来源土茯苓药材中落新妇苷和白藜芦醇的含量。方法:采用 YMC-PackODS-A(5 μm,4.6 mm×250 mm)色谱柱,乙腈(A)-0.05%磷酸水溶液(B)梯度洗脱(0～10 min 时25%A,10～15 min 时25%→35%A,15～20 min 时35%A);流速0.8 mL·min^(-1);检测波长306 nm。结果:落新妇苷、白藜芦醇浓度分别在0.018～0.288 mg·mL^(-1)和0.0025～0.025 mg·mL^(-1)范围内,与峰面积呈良好线性关系。落新妇苷平均回收率为99.6%～102.1%,RSD<1.9%;白藜芦醇平均回收率为99.2%～101.8%,RSD<2.2%。结论:该方法简便、准确,重复性好,可作为土茯苓药材的质量控制方法。
