A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference ...A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.展开更多
Aim To evaluate the anti-inflammatory effects of ethanol fraction prepared from Disporum cantoniense （Lour.） Merr. 70% ethanol extract with a cellular model of LPS-stimulated RAW264.7 cell. Methods RAW264.7 cells we...Aim To evaluate the anti-inflammatory effects of ethanol fraction prepared from Disporum cantoniense （Lour.） Merr. 70% ethanol extract with a cellular model of LPS-stimulated RAW264.7 cell. Methods RAW264.7 cells were treated with different concentrations of ethanol fraction （25,50 and 100 g · L^-1 ） and stimu- the conditioned media was collected and analyzed. The quantity of ni- lated with LPS （10 μg· L^-1） for 24 hours, tric oxide （NO） was assayed by Griess reagent. The production of inflammatory mediators was determined by en- zyme-linked immunosorbent assay （ELISA）, such as prostaglandin E2 （PGE2）, tumor necrosis factor ot （TNF-ot） interleukin- 1 β （IL-1 β） and interleukin 6 （IL-6） in cell supernatant. The concentrations of inflammatory medi- 9 ators were calculated according to the standard curves generated by each of the recombinant cytokines provided with LPS can induce RAW264.7 cells to promote the pro- the ELISA kits. Results Compared with the control group, TNF-α, IL-1β and IL-6. Compared with the duction of inflammatory mediators （P 〈 0.01 ） , including NO, PGE2, model group, ethanol fraction significantly suppressed LPS induced release of inflammatory mediators such as nitric NO, PGE2, TNF-α, IL-1β and IL-6 in a good dose dependent manner （P 〈 0.05, P 〈 0.01 ）. Conclusions Eth- anol fraction could significantly inhibit the production of LPS-induced inflammatory response in RAW264.7 cells, and its anti-inflammatory effect may be related to reduce the production of inflammatory mediators NO, PGE2, TNF-α, IL-1 β and IL-6. These results demonstrate that the ethanol fraction is the bioactive component of Disporum can- toniense （Lour.） Merr. , and the ethanol fraction will be further developed as a herbal remedy for preventive and/ or curative purposes in various inflammatory diseases.展开更多
基金Supported by Harbin Science and Technology Bureau(2016RQYXJ018,2017RAQXJ104)the Key Laboratory of Soybean Biology in the Chinese Ministry of Education,Northeast Agricultural University(SB17A01)+3 种基金the National Natural Science Foundation of China(31801386)Heilongjiang Natural Science Foundation(LC2018008)Heilongjiang General Young Innovative Talents Training Plan(UNPYSCT-2018158)Certificate of China Postdoctoral Science Foundation Grant(2018M641839)
文摘A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.
文摘Aim To evaluate the anti-inflammatory effects of ethanol fraction prepared from Disporum cantoniense （Lour.） Merr. 70% ethanol extract with a cellular model of LPS-stimulated RAW264.7 cell. Methods RAW264.7 cells were treated with different concentrations of ethanol fraction （25,50 and 100 g · L^-1 ） and stimu- the conditioned media was collected and analyzed. The quantity of ni- lated with LPS （10 μg· L^-1） for 24 hours, tric oxide （NO） was assayed by Griess reagent. The production of inflammatory mediators was determined by en- zyme-linked immunosorbent assay （ELISA）, such as prostaglandin E2 （PGE2）, tumor necrosis factor ot （TNF-ot） interleukin- 1 β （IL-1 β） and interleukin 6 （IL-6） in cell supernatant. The concentrations of inflammatory medi- 9 ators were calculated according to the standard curves generated by each of the recombinant cytokines provided with LPS can induce RAW264.7 cells to promote the pro- the ELISA kits. Results Compared with the control group, TNF-α, IL-1β and IL-6. Compared with the duction of inflammatory mediators （P 〈 0.01 ） , including NO, PGE2, model group, ethanol fraction significantly suppressed LPS induced release of inflammatory mediators such as nitric NO, PGE2, TNF-α, IL-1β and IL-6 in a good dose dependent manner （P 〈 0.05, P 〈 0.01 ）. Conclusions Eth- anol fraction could significantly inhibit the production of LPS-induced inflammatory response in RAW264.7 cells, and its anti-inflammatory effect may be related to reduce the production of inflammatory mediators NO, PGE2, TNF-α, IL-1 β and IL-6. These results demonstrate that the ethanol fraction is the bioactive component of Disporum can- toniense （Lour.） Merr. , and the ethanol fraction will be further developed as a herbal remedy for preventive and/ or curative purposes in various inflammatory diseases.
