Aim： To investigate antioxidant potential of lupeol/mango pulp extract （MPE） in testosterone induced oxidative stress in prostate of male Swiss albino mice. Methods： Oral treatment of lupeol （1 mg/animal） and M...Aim： To investigate antioxidant potential of lupeol/mango pulp extract （MPE） in testosterone induced oxidative stress in prostate of male Swiss albino mice. Methods： Oral treatment of lupeol （1 mg/animal） and MPE （1 mL [20% w/v]/ animal） was given separately to animals along with subcutaneous injection of testosterone （5 mg/kg body weight） consecutively for 15 days. At the end of the study period, the prostate was dissected out for the determination of reactive oxygen species （ROS） levels, lipid peroxidation and antioxidant enzymes status （catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase）. Results： In testosterone treated animals, increased ROS resulted in depletion of antioxidant enzymes and increase in lipid peroxidation in mouse prostate. However, lupeol/MPE treatment resulted in a decrease in ROS levels with restoration in the levels of lipid peroxidation and antioxidant enzymes. Conclusion： The results of the present study demonstrate that lupeol/MPE are effective in combating oxidative stress-induced cellular injury of mouse prostate. Mango and its constituents, therefore, deserve study as a potential chemopreventive agent against prostate cancer. （Asian J Androl 2008 Mar; 10： 313-318）展开更多
Objective:To optimize the ultrasonication method for efficient extraction ofβ-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology(RSM),and its valid...Objective:To optimize the ultrasonication method for efficient extraction ofβ-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology(RSM),and its validation by high performance thin layer chromatography(HPTLC)method.Methods:Ultrasonication method was used to extractβ-sitosterol and lupeol from Astragalus atropilosus(roots).RSM was used to optimize the different extraction parameters viz.liquid to solid ratio(10–14 m L/g),temperature(60-80℃)and time(40–60 min)to maximize the yield ofβ-sitosterol and lupeol.The quantitative estimation ofβ-sitosterol and lupeol was done in chloroform extract of Astragalus atropilosus by validated HPTLC method on 10 cm×20 cm glass-backed silica gel 60 F254 plate using hexane and ethyl acetate(8:2,v/v)as mobile phase.Results:A quadratic polynomial model was found to be most appropriate with regard to R1(yield of total extraction;R2/%CV=0.9948/0.28),R2(β-sitosterol yield;R2/%CV=0.9923/0.39)and R3(lupeol yield;R2/%CV=0.9942/0.97).The values of adjusted R2/predicted R2/signal to noise ratio for R1,R2,and R3 were 0.9782/0.9551/48.77,0.9904/0.9110/31.33,and 0.9927/0.9401/36.08,respectively,indicating a high degree of correlation and adequate signal.The linear correlation plot between the predicted and experimental values for R1,R2,and R3 showed high values of R2 ranging from 0.9905-0.9973.β-sitosterol and lupeol in chloroform extract of Astragalus atropilosus were detected at Rf values of 0.22 and 0.34,respectively,atλmax=518 nm.The optimized ultrasonic extraction produced 8.462%w/w of R1,0.451%w/w of R2 and 0.172%w/w of R3 at 13.5 m L/g liquid to solid ratio,78℃of temperature and 60 min of time.Conclusions:The experimental findings of RSM optimized extraction and HPTLC analysis can be further applied for the efficient extraction ofβ-sitosterol and lupeol in other species of Astragalus.展开更多
Objective: To discover lead lupane triterpenoid's potential isolated from Pueraria lobata roots against b-site amyloid precursor protein cleaving enzyme 1(BACE1), which serve as a rate limiting step in amyloid bet...Objective: To discover lead lupane triterpenoid's potential isolated from Pueraria lobata roots against b-site amyloid precursor protein cleaving enzyme 1(BACE1), which serve as a rate limiting step in amyloid beta(Aβ) production altering the course of Alzheimer's disease. In addition, enzyme kinetics study and molecular docking were conducted to establish the inhibition type and structure activity relationship.Methods: A systematic study of 70% ethanolic P. lobata root extract was employed to identify its BACE1 inhibitory potential. Further, BACE1 inhibitory potential of two lupane terpenoids, yielded from ethanolic extract, was assessed. In order to determine their inhibition mode, Lineweaver–Burk plots and Michaelis–Menten model for BACE1 was performed. Auto Dock 4.2 program in addition determined the molecular interaction of BACE1 with isolated terpenoids.Results: Considering the inhibitory potential of 70% ethanolic extract of P. lobata against BACE1(IC_(50)= 80.35 mg/mL), lupeol and lupenone were subsequently isolated and exhibited notable or moderate BACE1 inhibitory activity with IC_(50) values of 5.12 and 62.98 mmol/L, respectively, as compared to the positive control quercetin(IC_(50)= 21.28 mmol/L). The enzyme kinetics study enabled us to identify both compounds as competitive inhibitors, where lupeol displayed a very potent inhibition against BACE1 with low inhibition constant(Ki) value of 1.43 mmol/L, signifying greater binding affinity.In order to understand the binding mechanism and structure–activity relationship of two triterpene-based BACE1 inhibitors, we employed computer aided docking studies which evidently revealed that hydroxyl group of lupeol formed two hydrogen bonds with the ASP32(catalytic aspartic residue) and SER35 residues of BACE1 with the binding energy of(-8.2 kcal/mol), while the ketone group of lupenone did not form any hydrogen bonds with BACE1 giving evidence for less binding affinity. These results in turn have predicted the dependence of the inhibitory activity in the presence of hydroxyl group which has provided a new basis for BACE1 blockade.Conclusions: Our results have successfully explored the molecular mechanism of lupane triterpenoids via BACE1 inhibition, suggesting that lupeol in particular could be utilized as a useful therapeutic and preventive agent to mitigate Alzheimer's disease.展开更多
From the ethyl acetate extract of the medicinal plant Graptophyllum glandulosum Turrill, five known compounds: Lupeol (1), Oleanolic acid (2), Chrysoeriol (3), N-methyl-isonicotinamide (4) and β-sitosterol 3-O-β-D-g...From the ethyl acetate extract of the medicinal plant Graptophyllum glandulosum Turrill, five known compounds: Lupeol (1), Oleanolic acid (2), Chrysoeriol (3), N-methyl-isonicotinamide (4) and β-sitosterol 3-O-β-D-glucopyranoside (5) were isolated. In addition, oxidation reactions carried out on lupeol (1) yielded two semi-synthetic compounds, including a previously unreported: (20R)-formyloxy-29-nor-lupan-3-one (1b) and one other well-known Lupenone (1a). The structures of natural and semi-synthetic compounds were determined by analysis of 1D-(1H, 13C), 2D-(COSY, HSQC and HMBC) NMR data in conjunction with mass spectrometry (TOFESIMS and HR-TOFESIMS) and by comparison with the reported data. The evaluation of antimicrobial activities of substrate (1) as well as semi-synthetic derivatives (1a and 1b) using broth microdilution method showed that compound 1b was the most active (16 ≤ MIC ≤ 32 μg/mL) against Escherichia coli, Staphylococcus aureus and Candida albicans compared to the starting material 1 (16 ≤ MIC ≤ 64 μg/mL) and derivative 1a (32 ≤ MIC ≤ 64 μg/mL).展开更多
Objective:To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound,stigmasterol,in Abutilon indicum(A.indicum)using high-performance thin-layer ...Objective:To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound,stigmasterol,in Abutilon indicum(A.indicum)using high-performance thin-layer chromatography(HPTLC).Methods:TLC aluminum plates precoated with silica-gel 60 F254(20 cmí10 cm)were used with a mobile phase of toluene-methanol-formic acid(7.0:2.7:0.3,v/v/v)and densitometric determination of these compounds was carried out at 530 nm in reflectance/absorbance mode.Results:Compact bands for lupeol and stigmasterol were obtained at R_(f)0.52±0.02 and 0.28±0.05.The limit of detection(45 and 18 ng/band),limit of quantification(135 and 54 ng/band),recovery(98.2%-99.7%and 97.2%-99.6%)and precision(≤2.18 and 1.91)were satisfactory for lupeol and stigmasterol respectively.Linearity range for lupeol and stigmasterol were 100-1000(r^(2)=0.9994)and 50-500 ng/band(r^(2)=0.9941)and the contents were estimated as(0.59±0.10)%and(0.83±0.10)%w/w respectively.The total phenolic,flavonoid,proanthocyanidin,alkaloidal and saponin contents of methanolic extract of A.indicum were also measured in this work.According to International Conference on Harmonization(ICH)guidelines,the method was validated for linearity,precision,accuracy,and recovery,limit of detection,limit of quantification,specificity,and robustness.Conclusions:The HPTLC method was found to be reproducible,accurate,and precise and could detect these two compounds at nanogram level from the A.indicum.展开更多
文摘Aim： To investigate antioxidant potential of lupeol/mango pulp extract （MPE） in testosterone induced oxidative stress in prostate of male Swiss albino mice. Methods： Oral treatment of lupeol （1 mg/animal） and MPE （1 mL [20% w/v]/ animal） was given separately to animals along with subcutaneous injection of testosterone （5 mg/kg body weight） consecutively for 15 days. At the end of the study period, the prostate was dissected out for the determination of reactive oxygen species （ROS） levels, lipid peroxidation and antioxidant enzymes status （catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase）. Results： In testosterone treated animals, increased ROS resulted in depletion of antioxidant enzymes and increase in lipid peroxidation in mouse prostate. However, lupeol/MPE treatment resulted in a decrease in ROS levels with restoration in the levels of lipid peroxidation and antioxidant enzymes. Conclusion： The results of the present study demonstrate that lupeol/MPE are effective in combating oxidative stress-induced cellular injury of mouse prostate. Mango and its constituents, therefore, deserve study as a potential chemopreventive agent against prostate cancer. （Asian J Androl 2008 Mar; 10： 313-318）
基金the Researchers Supporting Project Number(RSP-2019/132),King Saud University,Riyadh,Kingdom of Saudi Arabia.
文摘Objective:To optimize the ultrasonication method for efficient extraction ofβ-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology(RSM),and its validation by high performance thin layer chromatography(HPTLC)method.Methods:Ultrasonication method was used to extractβ-sitosterol and lupeol from Astragalus atropilosus(roots).RSM was used to optimize the different extraction parameters viz.liquid to solid ratio(10–14 m L/g),temperature(60-80℃)and time(40–60 min)to maximize the yield ofβ-sitosterol and lupeol.The quantitative estimation ofβ-sitosterol and lupeol was done in chloroform extract of Astragalus atropilosus by validated HPTLC method on 10 cm×20 cm glass-backed silica gel 60 F254 plate using hexane and ethyl acetate(8:2,v/v)as mobile phase.Results:A quadratic polynomial model was found to be most appropriate with regard to R1(yield of total extraction;R2/%CV=0.9948/0.28),R2(β-sitosterol yield;R2/%CV=0.9923/0.39)and R3(lupeol yield;R2/%CV=0.9942/0.97).The values of adjusted R2/predicted R2/signal to noise ratio for R1,R2,and R3 were 0.9782/0.9551/48.77,0.9904/0.9110/31.33,and 0.9927/0.9401/36.08,respectively,indicating a high degree of correlation and adequate signal.The linear correlation plot between the predicted and experimental values for R1,R2,and R3 showed high values of R2 ranging from 0.9905-0.9973.β-sitosterol and lupeol in chloroform extract of Astragalus atropilosus were detected at Rf values of 0.22 and 0.34,respectively,atλmax=518 nm.The optimized ultrasonic extraction produced 8.462%w/w of R1,0.451%w/w of R2 and 0.172%w/w of R3 at 13.5 m L/g liquid to solid ratio,78℃of temperature and 60 min of time.Conclusions:The experimental findings of RSM optimized extraction and HPTLC analysis can be further applied for the efficient extraction ofβ-sitosterol and lupeol in other species of Astragalus.
基金Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(2012R1A6A1028677)
文摘Objective: To discover lead lupane triterpenoid's potential isolated from Pueraria lobata roots against b-site amyloid precursor protein cleaving enzyme 1(BACE1), which serve as a rate limiting step in amyloid beta(Aβ) production altering the course of Alzheimer's disease. In addition, enzyme kinetics study and molecular docking were conducted to establish the inhibition type and structure activity relationship.Methods: A systematic study of 70% ethanolic P. lobata root extract was employed to identify its BACE1 inhibitory potential. Further, BACE1 inhibitory potential of two lupane terpenoids, yielded from ethanolic extract, was assessed. In order to determine their inhibition mode, Lineweaver–Burk plots and Michaelis–Menten model for BACE1 was performed. Auto Dock 4.2 program in addition determined the molecular interaction of BACE1 with isolated terpenoids.Results: Considering the inhibitory potential of 70% ethanolic extract of P. lobata against BACE1(IC_(50)= 80.35 mg/mL), lupeol and lupenone were subsequently isolated and exhibited notable or moderate BACE1 inhibitory activity with IC_(50) values of 5.12 and 62.98 mmol/L, respectively, as compared to the positive control quercetin(IC_(50)= 21.28 mmol/L). The enzyme kinetics study enabled us to identify both compounds as competitive inhibitors, where lupeol displayed a very potent inhibition against BACE1 with low inhibition constant(Ki) value of 1.43 mmol/L, signifying greater binding affinity.In order to understand the binding mechanism and structure–activity relationship of two triterpene-based BACE1 inhibitors, we employed computer aided docking studies which evidently revealed that hydroxyl group of lupeol formed two hydrogen bonds with the ASP32(catalytic aspartic residue) and SER35 residues of BACE1 with the binding energy of(-8.2 kcal/mol), while the ketone group of lupenone did not form any hydrogen bonds with BACE1 giving evidence for less binding affinity. These results in turn have predicted the dependence of the inhibitory activity in the presence of hydroxyl group which has provided a new basis for BACE1 blockade.Conclusions: Our results have successfully explored the molecular mechanism of lupane triterpenoids via BACE1 inhibition, suggesting that lupeol in particular could be utilized as a useful therapeutic and preventive agent to mitigate Alzheimer's disease.
文摘From the ethyl acetate extract of the medicinal plant Graptophyllum glandulosum Turrill, five known compounds: Lupeol (1), Oleanolic acid (2), Chrysoeriol (3), N-methyl-isonicotinamide (4) and β-sitosterol 3-O-β-D-glucopyranoside (5) were isolated. In addition, oxidation reactions carried out on lupeol (1) yielded two semi-synthetic compounds, including a previously unreported: (20R)-formyloxy-29-nor-lupan-3-one (1b) and one other well-known Lupenone (1a). The structures of natural and semi-synthetic compounds were determined by analysis of 1D-(1H, 13C), 2D-(COSY, HSQC and HMBC) NMR data in conjunction with mass spectrometry (TOFESIMS and HR-TOFESIMS) and by comparison with the reported data. The evaluation of antimicrobial activities of substrate (1) as well as semi-synthetic derivatives (1a and 1b) using broth microdilution method showed that compound 1b was the most active (16 ≤ MIC ≤ 32 μg/mL) against Escherichia coli, Staphylococcus aureus and Candida albicans compared to the starting material 1 (16 ≤ MIC ≤ 64 μg/mL) and derivative 1a (32 ≤ MIC ≤ 64 μg/mL).
文摘Objective:To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound,stigmasterol,in Abutilon indicum(A.indicum)using high-performance thin-layer chromatography(HPTLC).Methods:TLC aluminum plates precoated with silica-gel 60 F254(20 cmí10 cm)were used with a mobile phase of toluene-methanol-formic acid(7.0:2.7:0.3,v/v/v)and densitometric determination of these compounds was carried out at 530 nm in reflectance/absorbance mode.Results:Compact bands for lupeol and stigmasterol were obtained at R_(f)0.52±0.02 and 0.28±0.05.The limit of detection(45 and 18 ng/band),limit of quantification(135 and 54 ng/band),recovery(98.2%-99.7%and 97.2%-99.6%)and precision(≤2.18 and 1.91)were satisfactory for lupeol and stigmasterol respectively.Linearity range for lupeol and stigmasterol were 100-1000(r^(2)=0.9994)and 50-500 ng/band(r^(2)=0.9941)and the contents were estimated as(0.59±0.10)%and(0.83±0.10)%w/w respectively.The total phenolic,flavonoid,proanthocyanidin,alkaloidal and saponin contents of methanolic extract of A.indicum were also measured in this work.According to International Conference on Harmonization(ICH)guidelines,the method was validated for linearity,precision,accuracy,and recovery,limit of detection,limit of quantification,specificity,and robustness.Conclusions:The HPTLC method was found to be reproducible,accurate,and precise and could detect these two compounds at nanogram level from the A.indicum.
文摘目的基于网络药理学和分子对接探讨羽扇豆醇(Lupeol)治疗类风湿关节炎(Rheumatoid arthritis,RA)的作用机制。方法通过Swiss和TCMSP分析平台获取羽扇豆醇的作用靶点,在GeneCards数据库检索“RA”获取其相关靶点,并转化为相应的标准化基因名。利用R包绘制羽扇豆醇和RA共同靶点的韦恩图,获取交集基因。将交集基因导入STRING数据库,构建蛋白-蛋白相互作用(Protein-protein interactions,PPI)网络。通过clusterProfiler数据库进行京都基因(Geneontology,GO)和基因组百科全书(Kyoto encyclopedia of genes and gnomes,KEGG)通路富集分析。通过分子对接评价羽扇豆醇与AR、CASP3和CCNB1的结合作用。构建RA小鼠模型,测量各组小鼠的足体积,HE染色检测病理学变化,ELISA试剂盒检测小鼠血清TNF-α、IL-1β和IL-6的表达水平,采用Western blot检测各组小鼠Bcl-2、Bax、Casp3、Casp9的蛋白表达水平。结果获取羽扇豆醇40个作用靶点,RA疾病4734个相关靶点,羽扇豆醇-RA共同靶点27个,PPI网络自由度靠前的3个基因为AR、CASP3和CCNB1,GO富集结果是291个和KEGG通路富集为20条信号通路。分子对接显示羽扇豆醇与AR、CASP3和CCNB1亲和作用较好。第8、12、16、20天模型组小鼠足体积显著高于正常对照组(P<0.05),相较于模型组,羽扇豆醇组在第8、12、16、20天时小鼠足体积显著降低(P<0.05),双氯芬酸钠组在第12、16、20天时小鼠足体积显著降低(P<0.05)。HE染色结果表明,与模型组相比,羽扇豆醇药物组明显改善病理学状态。与模型组相比,羽扇豆醇药物组小鼠血清中TNF-α、IL-1β和IL-6水平降低(P<0.01)。Western blot结果显示,与正常组相比,模型组小鼠Bax、Casp3和Casp9蛋白表达显著降低(P<0.05),Bcl-2的蛋白表达显著升高(P<0.05)。与模型组比较,双氯芬酸钠组和羽扇豆醇药物组小鼠Bax、Casp3和Casp9蛋白表达显著上调(P<0.05),Bcl-2蛋白表达显著下调(P<0.05)。结论羽扇豆醇可通过调控p53信号通路中的Bax、Bcl-2、Casp3和Casp9从而发挥对RA的治疗作用。
