Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic m...Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic medium （MS, WPM, B5 and N6）, cytokinins type and concentration （0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L 6-BA; 0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L KT）, auxins type and concentration （0, 0.25, 0.50, 0.75, 1.00 mg/L IBA, 2,4-D and NAA） on the induction of D. nipponica axillary buds were respectively measured and compared. Then, the effects of sugar source （30 g/L sucrose, fructose, white sugar, maltose and glucose） and light intensity （0, 800, 1 600, 2 400 and 3 200 lx） on the proliferation coefficient of D. nipponica axillary buds were evaluated.[Results] Using stem segments with leaf axils as the explants, the highest induction rate （90.8%） of axillary buds was achieved in MS supplemented with 2.5 mg/L 6-BA and 0.5 mg/L 2,4-D, and the fresh and dry weights of tissue culture seedlings in this medium were also the highest, up to 1.5 g and 160.9 mg, respectively. Then, the highest proliferation coefficient of was D. nipponica axillary buds noted when sucrose was used as the sugar source in medium, and the optimal light intensity for the proliferation of D. nipponica axillary buds was 2 400 lx.[Conclusions] The results provide an experimental evidence for rapid propagation of D. nipponica .展开更多
Objective To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino(TSDN)on the arachidonic acid pathway in monosodium urate(MSU)crystal-induced M1-polarized m...Objective To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino(TSDN)on the arachidonic acid pathway in monosodium urate(MSU)crystal-induced M1-polarized macrophages.Methods M1 polarization of RAW264.7 cells were induced by 1µg/mL lipopolysaccharide(LPS).The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN.MSU(500µg/mL)was used to induce the gouty arthritis model.Afterwards,10µg/L TSDN and 8µmol/L celecoxib,which was used as a positive control,were added to the above LPS and MSU-induced cells for 24 h.The mRNA and protein expressions of cyclooxygenase(COX)2,5-lipoxygenase(5-LOX),microsomal prostaglandin E synthase derived eicosanoids(mPGES)-1,leukotriene B(LTB)4,cytochrome P450(CYP)4A,and prostaglandin E_(2)(PGE_(2))were tested by real-time polymerase chain reaction and Western blotting,respectively.The enzyme-linked immunosorbent assay was used to test the contents of M1 markers,including inducible nitric oxid synthase(NOS)2,CD80,and CD86.Results TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2,5-LOX,CYP4A,LTB4,and PGE_(2)(P<0.01)while increased the mRNA and protein expression of mPGES-1(P<0.05 or P<0.01).TSDN could also significantly decrease the contents of NOS2,CD80,and CD86(P<0.01).Conclusion TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.展开更多
Objective: To investigate the mechanism of Chinese herbal medicine Dioscorea nipponica for the treatment of monosodium urate crystals-induced gouty arthritis(GA) in rats. Methods: Sixty male Wistar rats were divided i...Objective: To investigate the mechanism of Chinese herbal medicine Dioscorea nipponica for the treatment of monosodium urate crystals-induced gouty arthritis(GA) in rats. Methods: Sixty male Wistar rats were divided into 6 groups: normal, model, indomethacin and three total saponin(900, 300 and 100 mg/kg) groups. The liver, kidney and serum levels of lysosomal enzymes, antioxidant capacities, and inflammatory factors were measured. In addition, the m RNA and protein levels of the NALP3 inflammasome components in the mononuclear cells of rats’ peripheral blood were analyzed using real-time polymerase chain reaction and Western blotting methods, respectively. Results: Total saponins groups could reduce the activities of β-galactosidase,β-N acetyl glucosamine enzyme,β-glucuronidase, acid phosphatase, and malonaldehyde as wel as the contents of TNF-α, IL-1β and IL-8(all P<0.05). They could also increase the activities of glutathione peroxidase and total superoxide dismutase(both P<0.05). Further studies showed that total saponins groups of high, middle and low doses could all increase the m RNA and protein levels of caspase-1, adapter apoptosis-associated speck-like protein containing a caspase-recruitment domain(ASC) and NALP3 in the mononuclear cells of peripheral blood(all P<0.05). Conclusion: Dioscorea nipponica may treat GA by regulating lysosomal enzymes, antioxidant capacities and the NALP3 inflammasome.展开更多
Key for worldwide acceptance of Traditional Chinese Medicine(TCM)is the ability to provide scientific evidence combined with a quality control system based on the bioactive ingredients.Modern scientific technology too...Key for worldwide acceptance of Traditional Chinese Medicine(TCM)is the ability to provide scientific evidence combined with a quality control system based on the bioactive ingredients.Modern scientific technology tools are now available to accomplish standardization of TCM products in order to achieve a high level of efficacy and safety,enhancing the introduction into the展开更多
[目的]探讨穿山龙总皂苷对哮喘大鼠气道上皮细胞损伤的影响及对Janus激酶2(JAK2)/信号转导与转录激活子3(STAT3)信号通路的调节作用。[方法]采用卵清蛋白诱导建立哮喘大鼠模型,将建模成功的大鼠随机分为模型组、穿山龙总皂苷低剂量组、...[目的]探讨穿山龙总皂苷对哮喘大鼠气道上皮细胞损伤的影响及对Janus激酶2(JAK2)/信号转导与转录激活子3(STAT3)信号通路的调节作用。[方法]采用卵清蛋白诱导建立哮喘大鼠模型,将建模成功的大鼠随机分为模型组、穿山龙总皂苷低剂量组、穿山龙总皂苷中剂量组和穿山龙总皂苷高剂量组,另设正常对照组,支气管插管测定大鼠肺顺应性和肺弹性阻力,酶联免疫吸附法(ELISA)测定肺泡灌洗液中白介素(IL)-4、IL-6和干扰素γ(IFN-γ)水平,苏木精-伊红(HE)染色观察支气管病理损伤,实时荧光定量(qRT-PCR)法检测支气管组织中转化生长因子β1(TGF-β1)和Smad2 mRNA水平,蛋白印迹法检测p-JAK2和p-STAT3蛋白相对表达量。[结果]正常组大鼠肺组织结构正常完整,无炎性细胞浸润;模型组大鼠肺组织炎症中性粒细胞浸润、间质水肿、肺泡间隔增厚、肺泡内及间质斑片状出血;穿山龙低、中和高剂量组大鼠肺组织病理损伤减轻。与正常对照组比较,模型组大鼠肺顺应性降低,肺弹性阻力、IL-4、IL-6和IFN-γ水平、TGF-β1和Smad2 mRNA水平升高以及p-JAK2/JAK2和p-STAT3/STAT3升高(P<0.05);与模型组比较,穿山龙总皂苷低、中和高剂量组大鼠肺顺应性升高,肺弹性阻力,IL-4、IL-6和IFN-γ水平,TGF-β1和Smad2 m RNA水平降低以及p-JAK2/JAK2和p-STAT3/STAT3降低(P<0.05),其中各指标以穿山龙总皂苷高剂量组变化最为显著,其次为穿山龙中剂量组和穿山龙低剂量组(P<0.05)。[结论]穿山龙总皂苷可减轻哮喘大鼠气道上皮细胞损伤,降低炎症反应,改善气道重塑,增强肺顺应性,降低肺弹性阻力,其可能是通过抑制JAK2/STAT3信号通路发挥作用。展开更多
目的:以具有抗肿瘤活性的穿山龙乙酸乙酯层提取物(extract from Discorea nipponica Makino,DNM)和蒺藜提取物哈尔满碱为研究对象,采用体内裸鼠肿瘤HepG2模型实验探究两药联合应用是否具有协同抗肿瘤的作用。方法:通过建立裸鼠肿瘤HepG...目的:以具有抗肿瘤活性的穿山龙乙酸乙酯层提取物(extract from Discorea nipponica Makino,DNM)和蒺藜提取物哈尔满碱为研究对象,采用体内裸鼠肿瘤HepG2模型实验探究两药联合应用是否具有协同抗肿瘤的作用。方法:通过建立裸鼠肿瘤HepG2模型,灌胃给药两周,观察穿山龙乙酸乙酯层提取物组、哈尔满碱组和联合用药组的对肿瘤细胞的抑制情况;通过经统计学分析和采用金正均法计算分析验证结果;对肿瘤组织进行HE切片染色,观察联合用药对肿瘤组织中肿瘤细胞生长的影响。结果:穿山龙乙酸乙酯层提取物组、哈尔满碱组和联合用药组抑瘤率分别为3.36%、45.31%、59.59%,经统计学分析后有显著性差异(P<0.05),表明联合用药抑瘤率高于单独用药组。显微镜下观察联合用药组肿瘤组织出现大面积坏死,且出现肿瘤细胞核质比降低,异型性减小等现象。结论:说明穿山龙乙酸乙酯层提取物和蒺藜提取物哈尔满碱联合用药在裸鼠体内具有协同抗肿瘤的作用。展开更多
基金Supported by University-enterprise Cooperation Project of Yanbian University([2006]3)
文摘Objectives] This study aimed to determine the optimal medium components and culture conditions for the induction and proliferation of Dioscorea nipponica Makino axillary buds in vitro .[Methods] The effects of basic medium （MS, WPM, B5 and N6）, cytokinins type and concentration （0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L 6-BA; 0, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/L KT）, auxins type and concentration （0, 0.25, 0.50, 0.75, 1.00 mg/L IBA, 2,4-D and NAA） on the induction of D. nipponica axillary buds were respectively measured and compared. Then, the effects of sugar source （30 g/L sucrose, fructose, white sugar, maltose and glucose） and light intensity （0, 800, 1 600, 2 400 and 3 200 lx） on the proliferation coefficient of D. nipponica axillary buds were evaluated.[Results] Using stem segments with leaf axils as the explants, the highest induction rate （90.8%） of axillary buds was achieved in MS supplemented with 2.5 mg/L 6-BA and 0.5 mg/L 2,4-D, and the fresh and dry weights of tissue culture seedlings in this medium were also the highest, up to 1.5 g and 160.9 mg, respectively. Then, the highest proliferation coefficient of was D. nipponica axillary buds noted when sucrose was used as the sugar source in medium, and the optimal light intensity for the proliferation of D. nipponica axillary buds was 2 400 lx.[Conclusions] The results provide an experimental evidence for rapid propagation of D. nipponica .
基金Supported by Joint Guidance Project of Heilongjiang Natural Science Foundation(No.LH2021H099)Start Fund of Postdoctoral Research in Heilongjiang Province(No.LBH-Q19188)+3 种基金Outstanding Youth Development Fund of Heilongjiang University of Chinese Medicine(No.2019JC06)Project of Heilongjiang Administration of Traditional Chinese Medicine(No.ZHY202094)Graduate Innovative Scientific Research Project of Heilongjiang University of Chinese Medicine(No.2020yjscx057)Natural Science Foundation of Heilongjiang Province(Key project,No.ZD2020H006)。
文摘Objective To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino(TSDN)on the arachidonic acid pathway in monosodium urate(MSU)crystal-induced M1-polarized macrophages.Methods M1 polarization of RAW264.7 cells were induced by 1µg/mL lipopolysaccharide(LPS).The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN.MSU(500µg/mL)was used to induce the gouty arthritis model.Afterwards,10µg/L TSDN and 8µmol/L celecoxib,which was used as a positive control,were added to the above LPS and MSU-induced cells for 24 h.The mRNA and protein expressions of cyclooxygenase(COX)2,5-lipoxygenase(5-LOX),microsomal prostaglandin E synthase derived eicosanoids(mPGES)-1,leukotriene B(LTB)4,cytochrome P450(CYP)4A,and prostaglandin E_(2)(PGE_(2))were tested by real-time polymerase chain reaction and Western blotting,respectively.The enzyme-linked immunosorbent assay was used to test the contents of M1 markers,including inducible nitric oxid synthase(NOS)2,CD80,and CD86.Results TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2,5-LOX,CYP4A,LTB4,and PGE_(2)(P<0.01)while increased the mRNA and protein expression of mPGES-1(P<0.05 or P<0.01).TSDN could also significantly decrease the contents of NOS2,CD80,and CD86(P<0.01).Conclusion TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.
基金Supported by the National Natural Science Foundation of China(Nos.81173618,81403156)China Postdoctoral Science Foundation(No.2015M57036)+2 种基金Heilongjiang Postdoctoral Special Assisted Fund(Nos.LBH-TZ0520,LBH-Z13191)Scientific Fund of Heilongjiang University of Chinese Medicine(No.2013bs05)the Outstanding Innovative Talent Support Programs of Heilongjiang University of Chinese Medicine
文摘Objective: To investigate the mechanism of Chinese herbal medicine Dioscorea nipponica for the treatment of monosodium urate crystals-induced gouty arthritis(GA) in rats. Methods: Sixty male Wistar rats were divided into 6 groups: normal, model, indomethacin and three total saponin(900, 300 and 100 mg/kg) groups. The liver, kidney and serum levels of lysosomal enzymes, antioxidant capacities, and inflammatory factors were measured. In addition, the m RNA and protein levels of the NALP3 inflammasome components in the mononuclear cells of rats’ peripheral blood were analyzed using real-time polymerase chain reaction and Western blotting methods, respectively. Results: Total saponins groups could reduce the activities of β-galactosidase,β-N acetyl glucosamine enzyme,β-glucuronidase, acid phosphatase, and malonaldehyde as wel as the contents of TNF-α, IL-1β and IL-8(all P<0.05). They could also increase the activities of glutathione peroxidase and total superoxide dismutase(both P<0.05). Further studies showed that total saponins groups of high, middle and low doses could all increase the m RNA and protein levels of caspase-1, adapter apoptosis-associated speck-like protein containing a caspase-recruitment domain(ASC) and NALP3 in the mononuclear cells of peripheral blood(all P<0.05). Conclusion: Dioscorea nipponica may treat GA by regulating lysosomal enzymes, antioxidant capacities and the NALP3 inflammasome.
文摘Key for worldwide acceptance of Traditional Chinese Medicine(TCM)is the ability to provide scientific evidence combined with a quality control system based on the bioactive ingredients.Modern scientific technology tools are now available to accomplish standardization of TCM products in order to achieve a high level of efficacy and safety,enhancing the introduction into the
文摘[目的]探讨穿山龙总皂苷对哮喘大鼠气道上皮细胞损伤的影响及对Janus激酶2(JAK2)/信号转导与转录激活子3(STAT3)信号通路的调节作用。[方法]采用卵清蛋白诱导建立哮喘大鼠模型,将建模成功的大鼠随机分为模型组、穿山龙总皂苷低剂量组、穿山龙总皂苷中剂量组和穿山龙总皂苷高剂量组,另设正常对照组,支气管插管测定大鼠肺顺应性和肺弹性阻力,酶联免疫吸附法(ELISA)测定肺泡灌洗液中白介素(IL)-4、IL-6和干扰素γ(IFN-γ)水平,苏木精-伊红(HE)染色观察支气管病理损伤,实时荧光定量(qRT-PCR)法检测支气管组织中转化生长因子β1(TGF-β1)和Smad2 mRNA水平,蛋白印迹法检测p-JAK2和p-STAT3蛋白相对表达量。[结果]正常组大鼠肺组织结构正常完整,无炎性细胞浸润;模型组大鼠肺组织炎症中性粒细胞浸润、间质水肿、肺泡间隔增厚、肺泡内及间质斑片状出血;穿山龙低、中和高剂量组大鼠肺组织病理损伤减轻。与正常对照组比较,模型组大鼠肺顺应性降低,肺弹性阻力、IL-4、IL-6和IFN-γ水平、TGF-β1和Smad2 mRNA水平升高以及p-JAK2/JAK2和p-STAT3/STAT3升高(P<0.05);与模型组比较,穿山龙总皂苷低、中和高剂量组大鼠肺顺应性升高,肺弹性阻力,IL-4、IL-6和IFN-γ水平,TGF-β1和Smad2 m RNA水平降低以及p-JAK2/JAK2和p-STAT3/STAT3降低(P<0.05),其中各指标以穿山龙总皂苷高剂量组变化最为显著,其次为穿山龙中剂量组和穿山龙低剂量组(P<0.05)。[结论]穿山龙总皂苷可减轻哮喘大鼠气道上皮细胞损伤,降低炎症反应,改善气道重塑,增强肺顺应性,降低肺弹性阻力,其可能是通过抑制JAK2/STAT3信号通路发挥作用。
文摘目的:以具有抗肿瘤活性的穿山龙乙酸乙酯层提取物(extract from Discorea nipponica Makino,DNM)和蒺藜提取物哈尔满碱为研究对象,采用体内裸鼠肿瘤HepG2模型实验探究两药联合应用是否具有协同抗肿瘤的作用。方法:通过建立裸鼠肿瘤HepG2模型,灌胃给药两周,观察穿山龙乙酸乙酯层提取物组、哈尔满碱组和联合用药组的对肿瘤细胞的抑制情况;通过经统计学分析和采用金正均法计算分析验证结果;对肿瘤组织进行HE切片染色,观察联合用药对肿瘤组织中肿瘤细胞生长的影响。结果:穿山龙乙酸乙酯层提取物组、哈尔满碱组和联合用药组抑瘤率分别为3.36%、45.31%、59.59%,经统计学分析后有显著性差异(P<0.05),表明联合用药抑瘤率高于单独用药组。显微镜下观察联合用药组肿瘤组织出现大面积坏死,且出现肿瘤细胞核质比降低,异型性减小等现象。结论:说明穿山龙乙酸乙酯层提取物和蒺藜提取物哈尔满碱联合用药在裸鼠体内具有协同抗肿瘤的作用。
