It is necessary to explore potent therapeutic agents via regulating gut microbiota and metabolism to combat Parkinson's disease(PD).Dioscin,a bioactive steroidal saponin,shows various activities.However,its effect...It is necessary to explore potent therapeutic agents via regulating gut microbiota and metabolism to combat Parkinson's disease(PD).Dioscin,a bioactive steroidal saponin,shows various activities.However,its effects and mechanisms against PD are limited.In this study,dioscin dramatically alleviated neuroinflammation and oxidative stress,and restored the disorders of mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).16 S rDNA sequencing assay demonstrated that dioscin reversed MPTP-induced gut dysbiosis to decrease Firmicutes-to-Bacteroidetes ratio and the abundances of Enterococcus,Streptococcus,Bacteroides and Lactobacillus genera,which further inhibited bile salt hydrolase(BSH)activity and blocked bile acid(BA)deconjugation.Fecal microbiome transplantation test showed that the anti-PD effect of dioscin was gut microbiota-dependent.In addition,non-targeted fecal metabolomics assays revealed many differential metabolites in adjusting steroid biosynthesis and primary bile acid biosynthesis.Moreover,targeted bile acid metabolomics assay indicated that dioscin increased the levels of ursodeoxycholic acid,tauroursodeoxycholic acid,taurodeoxycholic acid and bmuricholic acid in feces and serum.In addition,ursodeoxycholic acid administration markedly improved the protective effects of dioscin against PD in mice.Mechanistic test indicated that dioscin significantly up-regulated the levels of takeda G protein-coupled receptor 5(TGR5),glucagon-like peptide-1 receptor(GLP-1R),GLP-1,superoxide dismutase(SOD),and down-regulated NADPH oxidases 2(NOX2)and nuclear factor-kappaB(NF-kB)levels.Our data indicated that dioscin ameliorated PD phenotype by restoring gut dysbiosis and regulating bile acid-mediated oxidative stress and neuroinflammation via targeting GLP-1 signal in MPTP-induced PD mice,suggesting that the compound should be considered as a prebiotic agent to treat PD in the future.展开更多
Background and Objective:LTB4 has been shown to be involved in rheumatoid arthritis(RA)pathogenesis.The effect of Dioscin(Dio)on the LTB4 pathway of RA have not been reported yet.This study aimed at further exploring ...Background and Objective:LTB4 has been shown to be involved in rheumatoid arthritis(RA)pathogenesis.The effect of Dioscin(Dio)on the LTB4 pathway of RA have not been reported yet.This study aimed at further exploring whether Dioscin’s effects on TNF-αinduced collagen-induced arthritis(CIA)rat fibroblast-like synoviocytes(FLS)connected with the LTB4 and its receptor pathway.Materials&Methods:In this experiment,control group,TNF-αgroup,and different concentrations of Dioscin groups were established.Cell viability was evaluated using MTT assay.The levels of LTB4 in the samples of above groups were measured using ELISA.The mRNA expression levels of LTA4H,BLT1,and BLT2 were detected by quantitative real time PCR,while the expression level of LTA4H proteins were detected using western blot.The distribution of LTA4H was assessed by immunofluorescence assay.Results:the LTB4 level of TNF-αgroup in sample supernatant was higher than both control group and Dioscin groups with decreased LTB4 levels(p<0.05).Compared with the control group,the expression of LTA4H was significantly increased in TNF-αgroup(p<0.05),whereas LTA4H expressions were significantly decreased in all Dioscin groups when compared to TNF-αgroup(p<0.05).The mRNA expressions of BLT1 and BLT2 were markedly higher in TNF-αgroup than those in control group while Dioscin treatment significantly inhibited the increased expressions of BLT1 and BLT2 induced by TNF-α(p<0.05).Conclusions:These results firstly demonstrate that the protective effect of Dioscin on TNF-αinduced FLS may involve in its reducing LTB4 production by down-regulating LTA4H expression,and may inhibit its downstream pathway by decreasing LTB4 receptors levels.This findings suggest that dioscin produces a potential therapeutic effects for RA via its influencing LTA4H/LTB4/BLT pathway.展开更多
Dioscin is a natural steroid saponin derived from several plants,showing potent anti-cancer effect against a variety of tumor cell lines.In the present study,we investigated the anti-cancer activity of dioscin against...Dioscin is a natural steroid saponin derived from several plants,showing potent anti-cancer effect against a variety of tumor cell lines.In the present study,we investigated the anti-cancer activity of dioscin against human LNCaP cells,and evaluated the possible mechanism involved in its antineoplastic action.It was found that dioscin(1,2 and 4μmol/L)could significantly inhibit the viability of LNCaP cells in a time-and concentration-dependent manner.Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h,indicating that apoptosis was an important mechanism by which dioscin inhibited cancer.Western blotting was employed to detect the expression of caspase-3,Bcl-2 and Bax in LNCaP cells.The expression of cleaved caspase-3 was significantly increased,and meanwhile procaspase-3 was markedly decreased.The expression of anti-apoptotic protein Bcl-2 was down-regulated,whereas the pro-apoptotic protein Bax was up-regulated.Moreover,the Bcl-2/Bax ratio was drastically decreased.These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway,which might be associated with caspase-3 and Bcl-2 protein family.展开更多
Dioscin was extracted and isolated from Polygonatum Zanlanscianense Pamp. The effects of dioscin on HL60, HeLa, H14, and MDA MB 435 cell lines were studied with the results showing that dioscin dramatically inhibited ...Dioscin was extracted and isolated from Polygonatum Zanlanscianense Pamp. The effects of dioscin on HL60, HeLa, H14, and MDA MB 435 cell lines were studied with the results showing that dioscin dramatically inhibited the growth of the MDA MB 435, H14, HL60, and HeLa cell lines. The IC 50 of dioscin on these cell lines were 2.6, 0.8, 7.5, and 4.5 μmol/L respectively.展开更多
目的探讨薯蓣皂苷(Dioscin)通过调控腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)自噬信号通路对人骨肉瘤MG63细胞增殖及侵袭的影响。方法取对数期生长人骨肉瘤MG63细胞分为空白对照组、Dioscin不同剂量组(20、40、80、100μ...目的探讨薯蓣皂苷(Dioscin)通过调控腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)自噬信号通路对人骨肉瘤MG63细胞增殖及侵袭的影响。方法取对数期生长人骨肉瘤MG63细胞分为空白对照组、Dioscin不同剂量组(20、40、80、100μmol/L)。CCK-8法检测细胞增殖能力,划痕实验检测细胞迁移能力,细胞迁移实验检测细胞侵袭能力,丹酰尸胺法观察细胞自噬情况,蛋白质印迹法检测磷酸化腺苷酸活化蛋白激酶(p-AMPK)/AMPK、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)/mTOR、转录因子EB(TFEB)、人微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)、自噬效应蛋白beclin-1表达水平。结果与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞增殖率[(65.21±4.09)%、(51.94±3.95)%、(46.57±3.72)%、(37.31±4.23)%比100.00%]、划痕愈合率[(85.43±3.84)%、(70.62±3.17)%、(55.83±2.92)%、(44.23±3.27)%比(98.25±1.23)%]、侵袭细胞比率[(72.43±2.83)%、(63.76±3.34)%、(51.92±3.13)%、(42.44±3.09)%比(91.14±2.91)%]依次降低,呈浓度依赖性(P<0.05);空白对照组中仅有少量细胞自噬泡,作用48 h后Dioscin各组自噬泡依次增多;与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞中p-AMPK/AMPK[(0.35±0.04)、(0.50±0.02)、(0.60±0.03)、(0.88±0.03)比(0.20±0.02)]、TFEB[(0.42±0.02)、(0.68±0.03)、(0.92±0.04)、(1.36±0.10)比(0.30±0.02)]、LC3-Ⅱ[(0.41±0.03)、(0.70±0.04)、(0.88±0.03)、(1.17±0.07)比(0.26±0.02)]、beclin-1[(0.32±0.03)、(0.50±0.04)、(0.68±0.05)、(0.75±0.06)比(0.16±0.02)]蛋白表达依次增加,p-mTOR/mTOR[(0.75±0.02)、(0.52±0.03)、(0.40±0.04)、(0.31±0.02)比(0.86±0.05)]蛋白表达依次降低,差异均具有统计学意义(P<0.05);与Dioscin(100μmol/L)组比较,Dioscin(100μmol/L)+AMPK抑制剂(Compound C 10μmol/L)组p-AMPK/AMPK[(0.49±0.03)比(0.63±0.04)]、TFEB[(0.16±0.03)比(0.24±0.02)]、LC3-Ⅱ[(0.35±0.02)比(0.44±0.04)]、beclin-1[(0.44±0.03)比(0.56±0.04)]蛋白表达水平明显降低,p-mTOR/mTOR[(0.63±0.05)比(0.45±0.04)]表达明显升高(P<0.05)。结论Dioscin可能通过调控AMPK/mTOR-TFEB通路抑制骨肉瘤细胞增殖、迁移及侵袭。展开更多
基金funding from the Spring City Plan:The High-Level Talent Promotion and Training Project of Kunming and the Independent Research Fund of Yunnan Characteristic Plant Extraction Laboratory(Grant No.:2022YKZY001).
文摘It is necessary to explore potent therapeutic agents via regulating gut microbiota and metabolism to combat Parkinson's disease(PD).Dioscin,a bioactive steroidal saponin,shows various activities.However,its effects and mechanisms against PD are limited.In this study,dioscin dramatically alleviated neuroinflammation and oxidative stress,and restored the disorders of mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).16 S rDNA sequencing assay demonstrated that dioscin reversed MPTP-induced gut dysbiosis to decrease Firmicutes-to-Bacteroidetes ratio and the abundances of Enterococcus,Streptococcus,Bacteroides and Lactobacillus genera,which further inhibited bile salt hydrolase(BSH)activity and blocked bile acid(BA)deconjugation.Fecal microbiome transplantation test showed that the anti-PD effect of dioscin was gut microbiota-dependent.In addition,non-targeted fecal metabolomics assays revealed many differential metabolites in adjusting steroid biosynthesis and primary bile acid biosynthesis.Moreover,targeted bile acid metabolomics assay indicated that dioscin increased the levels of ursodeoxycholic acid,tauroursodeoxycholic acid,taurodeoxycholic acid and bmuricholic acid in feces and serum.In addition,ursodeoxycholic acid administration markedly improved the protective effects of dioscin against PD in mice.Mechanistic test indicated that dioscin significantly up-regulated the levels of takeda G protein-coupled receptor 5(TGR5),glucagon-like peptide-1 receptor(GLP-1R),GLP-1,superoxide dismutase(SOD),and down-regulated NADPH oxidases 2(NOX2)and nuclear factor-kappaB(NF-kB)levels.Our data indicated that dioscin ameliorated PD phenotype by restoring gut dysbiosis and regulating bile acid-mediated oxidative stress and neuroinflammation via targeting GLP-1 signal in MPTP-induced PD mice,suggesting that the compound should be considered as a prebiotic agent to treat PD in the future.
基金supported by the National Natural Science Foundation of China(Nos.82060661,81660751,81660151)Jiangxi Provincial Natural Science Foundation of China(No.20171BAB205085).
文摘Background and Objective:LTB4 has been shown to be involved in rheumatoid arthritis(RA)pathogenesis.The effect of Dioscin(Dio)on the LTB4 pathway of RA have not been reported yet.This study aimed at further exploring whether Dioscin’s effects on TNF-αinduced collagen-induced arthritis(CIA)rat fibroblast-like synoviocytes(FLS)connected with the LTB4 and its receptor pathway.Materials&Methods:In this experiment,control group,TNF-αgroup,and different concentrations of Dioscin groups were established.Cell viability was evaluated using MTT assay.The levels of LTB4 in the samples of above groups were measured using ELISA.The mRNA expression levels of LTA4H,BLT1,and BLT2 were detected by quantitative real time PCR,while the expression level of LTA4H proteins were detected using western blot.The distribution of LTA4H was assessed by immunofluorescence assay.Results:the LTB4 level of TNF-αgroup in sample supernatant was higher than both control group and Dioscin groups with decreased LTB4 levels(p<0.05).Compared with the control group,the expression of LTA4H was significantly increased in TNF-αgroup(p<0.05),whereas LTA4H expressions were significantly decreased in all Dioscin groups when compared to TNF-αgroup(p<0.05).The mRNA expressions of BLT1 and BLT2 were markedly higher in TNF-αgroup than those in control group while Dioscin treatment significantly inhibited the increased expressions of BLT1 and BLT2 induced by TNF-α(p<0.05).Conclusions:These results firstly demonstrate that the protective effect of Dioscin on TNF-αinduced FLS may involve in its reducing LTB4 production by down-regulating LTA4H expression,and may inhibit its downstream pathway by decreasing LTB4 receptors levels.This findings suggest that dioscin produces a potential therapeutic effects for RA via its influencing LTA4H/LTB4/BLT pathway.
基金supported by the National Natural Science Foundation of China(No.81173065)
文摘Dioscin is a natural steroid saponin derived from several plants,showing potent anti-cancer effect against a variety of tumor cell lines.In the present study,we investigated the anti-cancer activity of dioscin against human LNCaP cells,and evaluated the possible mechanism involved in its antineoplastic action.It was found that dioscin(1,2 and 4μmol/L)could significantly inhibit the viability of LNCaP cells in a time-and concentration-dependent manner.Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h,indicating that apoptosis was an important mechanism by which dioscin inhibited cancer.Western blotting was employed to detect the expression of caspase-3,Bcl-2 and Bax in LNCaP cells.The expression of cleaved caspase-3 was significantly increased,and meanwhile procaspase-3 was markedly decreased.The expression of anti-apoptotic protein Bcl-2 was down-regulated,whereas the pro-apoptotic protein Bax was up-regulated.Moreover,the Bcl-2/Bax ratio was drastically decreased.These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway,which might be associated with caspase-3 and Bcl-2 protein family.
基金partly by the Tsinghua U niversity- HongKong Baptist U niversity Joint Institute for Research ofChinese Medicine the" 985" Fund fromTsinghua U niversity.
文摘Dioscin was extracted and isolated from Polygonatum Zanlanscianense Pamp. The effects of dioscin on HL60, HeLa, H14, and MDA MB 435 cell lines were studied with the results showing that dioscin dramatically inhibited the growth of the MDA MB 435, H14, HL60, and HeLa cell lines. The IC 50 of dioscin on these cell lines were 2.6, 0.8, 7.5, and 4.5 μmol/L respectively.
文摘目的探讨薯蓣皂苷(Dioscin)通过调控腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)自噬信号通路对人骨肉瘤MG63细胞增殖及侵袭的影响。方法取对数期生长人骨肉瘤MG63细胞分为空白对照组、Dioscin不同剂量组(20、40、80、100μmol/L)。CCK-8法检测细胞增殖能力,划痕实验检测细胞迁移能力,细胞迁移实验检测细胞侵袭能力,丹酰尸胺法观察细胞自噬情况,蛋白质印迹法检测磷酸化腺苷酸活化蛋白激酶(p-AMPK)/AMPK、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)/mTOR、转录因子EB(TFEB)、人微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)、自噬效应蛋白beclin-1表达水平。结果与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞增殖率[(65.21±4.09)%、(51.94±3.95)%、(46.57±3.72)%、(37.31±4.23)%比100.00%]、划痕愈合率[(85.43±3.84)%、(70.62±3.17)%、(55.83±2.92)%、(44.23±3.27)%比(98.25±1.23)%]、侵袭细胞比率[(72.43±2.83)%、(63.76±3.34)%、(51.92±3.13)%、(42.44±3.09)%比(91.14±2.91)%]依次降低,呈浓度依赖性(P<0.05);空白对照组中仅有少量细胞自噬泡,作用48 h后Dioscin各组自噬泡依次增多;与空白对照组比较,Dioscin20、40、80、100μmol/L组MG63细胞中p-AMPK/AMPK[(0.35±0.04)、(0.50±0.02)、(0.60±0.03)、(0.88±0.03)比(0.20±0.02)]、TFEB[(0.42±0.02)、(0.68±0.03)、(0.92±0.04)、(1.36±0.10)比(0.30±0.02)]、LC3-Ⅱ[(0.41±0.03)、(0.70±0.04)、(0.88±0.03)、(1.17±0.07)比(0.26±0.02)]、beclin-1[(0.32±0.03)、(0.50±0.04)、(0.68±0.05)、(0.75±0.06)比(0.16±0.02)]蛋白表达依次增加,p-mTOR/mTOR[(0.75±0.02)、(0.52±0.03)、(0.40±0.04)、(0.31±0.02)比(0.86±0.05)]蛋白表达依次降低,差异均具有统计学意义(P<0.05);与Dioscin(100μmol/L)组比较,Dioscin(100μmol/L)+AMPK抑制剂(Compound C 10μmol/L)组p-AMPK/AMPK[(0.49±0.03)比(0.63±0.04)]、TFEB[(0.16±0.03)比(0.24±0.02)]、LC3-Ⅱ[(0.35±0.02)比(0.44±0.04)]、beclin-1[(0.44±0.03)比(0.56±0.04)]蛋白表达水平明显降低,p-mTOR/mTOR[(0.63±0.05)比(0.45±0.04)]表达明显升高(P<0.05)。结论Dioscin可能通过调控AMPK/mTOR-TFEB通路抑制骨肉瘤细胞增殖、迁移及侵袭。
