This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Ta...This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets.Injury-related molecules were obtained from GeenCards database,and the predicted targets of PF for injury treatment were selected by Wayne diagram.For mechanism analysis,the protein-protein interactions were constructed by String,and the KEGG analysis was conducted in Webgestalt.Then,cell viability and cytotoxicity assay were established by CCK8 assay.Also,the experimental cells were allocated to control,model(200μmol·L^(−1) H_(2)O_(2)),SB20358010μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+SB20358010μmol·L^(−1)),PF 50μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 50μmol·L^(−1)),and PF 100μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 100μmol·L^(−1))groups.We measured the intracellular ROS,Hoechst 33258 staining,cell apoptosis,the levels of Bcl-xl,Bcl-2,Caspase-3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK.There are 96 potential targets that may be associated with PF for injury treatment.Then,we chose the“Inflammatory mediator regulation of TRP channels”pathway for the experimental verification from the first 10 KEGG pathway.In experimental verification,H_(2)O_(2) decreased the cell viability moderately(P<0.05),and 100μmol·L^(−1) PF increased the cell viability significantly(P<0.05).Depending on the difference of intracellular ROS fluorescence intensity,PF inhibited H_(2)O_(2)-induced reactive oxygen species production in Schwann cells.In Hoechst 33258 staining,PF reversed the condensed chromatin and apoptotic nuclei following H_(2)O_(2) treatment.Moreover,Flow cytometry results showed that PF could substantially inhibit H_(2)O_(2) induced apoptosis(P<0.05).Pretreatment with PF obviously reduced the levels of Caspase3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the展开更多
生长停滞特异性蛋白6(growth arrest specific protein 6,GAS6)在肿瘤发生发展中发挥重要作用,其信号转导参与细胞增殖、黏附与迁移,但它在子宫内膜异位症(endometriosis, EMs)的相关功能及分子机制尚不明确。本研究从GEO数据库检索并...生长停滞特异性蛋白6(growth arrest specific protein 6,GAS6)在肿瘤发生发展中发挥重要作用,其信号转导参与细胞增殖、黏附与迁移,但它在子宫内膜异位症(endometriosis, EMs)的相关功能及分子机制尚不明确。本研究从GEO数据库检索并下载子宫内膜异位症相关转录物组数据集,并对其进行GEO在线分析,筛选差异表达基因并进行GO聚类和KEGG通路富集分析。利用10例无内异症且无明确疾病妇女的在位子宫内膜,以及11例卵巢巧克力囊肿病人异位子宫内膜,对3个以上数据集共有的差异基因的mRNA水平进行实时荧光定量PCR验证。在子宫内膜异位症临床样本中,采用免疫组化、实时荧光定量PCR验证关键调控因子GAS6及上皮间充质转化(epithelial mesenchymal transition, EMT)标记基因的表达水平,并利用免疫荧光对GAS6和E-钙黏着蛋白(E-cadherin)进行共标。研究发现:从4个转录物组数据集中共筛选出47个差异表达基因,其主要富集于细胞迁移等过程以及MAPK、PI3K-AKT、紧密连接等相关信号通路。3个以上数据集所共有的9个差异基因在子宫内膜异位症病人中的mRNA水平均符合生物信息学分析的结果。GAS6在子宫内膜异位症病人异位内膜中的表达水平高于对照组(P<0.05),并且子宫内膜异位症病人的内膜组织中存在EMT现象,EMT的标志物E-钙黏着蛋白表达水平下调(P<0.05)、波形蛋白(vimentin)表达水平上调(P<0.01)。在GAS6高表达的子宫内膜异位症患者异位子宫内膜腺上皮细胞中,E-cadherin显示低表达,提示GAS6可能在子宫内膜异位症中介导EMT过程。综上所述,本研究初步揭示GAS6在子宫内膜异位症病人中高表达,及其可能介导EMT过程参与子宫内膜异位症的发生与发展,为子宫内膜异位症的临床治疗提供潜在靶标。展开更多
Tillering is an important agronomic trait which has a direct impact on plant type and grain yield. Strigolactones are a class of important phytohormones regulating rice tillering. ATMAX1 is an important gene involved ...Tillering is an important agronomic trait which has a direct impact on plant type and grain yield. Strigolactones are a class of important phytohormones regulating rice tillering. ATMAX1 is an important gene involved in strigolactone biosynthesis through encoding the protein P450 in Arabidopsis. Based on sequence BLASTp, we identified five homologous genes of ATMAX1 in rice, i.e., OsMAXla, OsMAXlb, OsMAXlc, OsMAXld and OsMAXle. Among them, OsMAXla and OsMAXle showed stable and high expression in rice tissues. In addition, we observed that OsMAXla and OsMAXle can rescue the branched phenotype and the influences caused by MAX1 mutation in Arabidopsis. Moreover, the expression of OsMAX1a and OsMAXle can respond to phosphate deficiency and different phytohormones, especially GR24, a strigolactone analogue. Therefore, it is concluded that OsMAX1a and OsMAX1e are involved in the biosynthesis of strigolactones and regulated rice tillering.展开更多
基金the National Natural Science Foundation of China(No.81874404).
文摘This study was to investigate the protective effect of paeoniflorin(PF)on hydrogen peroxide-induced injury.Firstly,“SMILES”of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets.Injury-related molecules were obtained from GeenCards database,and the predicted targets of PF for injury treatment were selected by Wayne diagram.For mechanism analysis,the protein-protein interactions were constructed by String,and the KEGG analysis was conducted in Webgestalt.Then,cell viability and cytotoxicity assay were established by CCK8 assay.Also,the experimental cells were allocated to control,model(200μmol·L^(−1) H_(2)O_(2)),SB20358010μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+SB20358010μmol·L^(−1)),PF 50μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 50μmol·L^(−1)),and PF 100μmol·L^(−1)(200μmol·L^(−1) H_(2)O_(2)+PF 100μmol·L^(−1))groups.We measured the intracellular ROS,Hoechst 33258 staining,cell apoptosis,the levels of Bcl-xl,Bcl-2,Caspase-3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the phosphorylation expression of p38MAPK.There are 96 potential targets that may be associated with PF for injury treatment.Then,we chose the“Inflammatory mediator regulation of TRP channels”pathway for the experimental verification from the first 10 KEGG pathway.In experimental verification,H_(2)O_(2) decreased the cell viability moderately(P<0.05),and 100μmol·L^(−1) PF increased the cell viability significantly(P<0.05).Depending on the difference of intracellular ROS fluorescence intensity,PF inhibited H_(2)O_(2)-induced reactive oxygen species production in Schwann cells.In Hoechst 33258 staining,PF reversed the condensed chromatin and apoptotic nuclei following H_(2)O_(2) treatment.Moreover,Flow cytometry results showed that PF could substantially inhibit H_(2)O_(2) induced apoptosis(P<0.05).Pretreatment with PF obviously reduced the levels of Caspase3,Cleaved-caspase3,Cleaved-caspase7,TRPA1,TRPV1,and the
文摘生长停滞特异性蛋白6(growth arrest specific protein 6,GAS6)在肿瘤发生发展中发挥重要作用,其信号转导参与细胞增殖、黏附与迁移,但它在子宫内膜异位症(endometriosis, EMs)的相关功能及分子机制尚不明确。本研究从GEO数据库检索并下载子宫内膜异位症相关转录物组数据集,并对其进行GEO在线分析,筛选差异表达基因并进行GO聚类和KEGG通路富集分析。利用10例无内异症且无明确疾病妇女的在位子宫内膜,以及11例卵巢巧克力囊肿病人异位子宫内膜,对3个以上数据集共有的差异基因的mRNA水平进行实时荧光定量PCR验证。在子宫内膜异位症临床样本中,采用免疫组化、实时荧光定量PCR验证关键调控因子GAS6及上皮间充质转化(epithelial mesenchymal transition, EMT)标记基因的表达水平,并利用免疫荧光对GAS6和E-钙黏着蛋白(E-cadherin)进行共标。研究发现:从4个转录物组数据集中共筛选出47个差异表达基因,其主要富集于细胞迁移等过程以及MAPK、PI3K-AKT、紧密连接等相关信号通路。3个以上数据集所共有的9个差异基因在子宫内膜异位症病人中的mRNA水平均符合生物信息学分析的结果。GAS6在子宫内膜异位症病人异位内膜中的表达水平高于对照组(P<0.05),并且子宫内膜异位症病人的内膜组织中存在EMT现象,EMT的标志物E-钙黏着蛋白表达水平下调(P<0.05)、波形蛋白(vimentin)表达水平上调(P<0.01)。在GAS6高表达的子宫内膜异位症患者异位子宫内膜腺上皮细胞中,E-cadherin显示低表达,提示GAS6可能在子宫内膜异位症中介导EMT过程。综上所述,本研究初步揭示GAS6在子宫内膜异位症病人中高表达,及其可能介导EMT过程参与子宫内膜异位症的发生与发展,为子宫内膜异位症的临床治疗提供潜在靶标。
文摘Tillering is an important agronomic trait which has a direct impact on plant type and grain yield. Strigolactones are a class of important phytohormones regulating rice tillering. ATMAX1 is an important gene involved in strigolactone biosynthesis through encoding the protein P450 in Arabidopsis. Based on sequence BLASTp, we identified five homologous genes of ATMAX1 in rice, i.e., OsMAXla, OsMAXlb, OsMAXlc, OsMAXld and OsMAXle. Among them, OsMAXla and OsMAXle showed stable and high expression in rice tissues. In addition, we observed that OsMAXla and OsMAXle can rescue the branched phenotype and the influences caused by MAX1 mutation in Arabidopsis. Moreover, the expression of OsMAX1a and OsMAXle can respond to phosphate deficiency and different phytohormones, especially GR24, a strigolactone analogue. Therefore, it is concluded that OsMAX1a and OsMAX1e are involved in the biosynthesis of strigolactones and regulated rice tillering.
