Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human or...Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human oral mucosafibroblasts(hOMF)were induced with transforming growth factor beta1(TGF-β1)and intervened with Pue.Expressions offibrosis-related markers were analyzed by Western blot and IF staining.Cell viability was characterized by the CCK-8 assay.Expressions of miR-30 family members were quantified by qRT-PCR.The correlation betweenfibroblast activation protein(FAP)and miR-30 family expression was evaluated by the Pearson correlation coefficient.Bioinformatics prediction and dual-luciferase reporter assay were employed to verify the regulation between FAP and miR-30b-5p.The specific mechanism of Pue on OSF was explored through the promotion or inhibition of miR-30b-5p.Results:After induction by TGF-β1,hOMF showed upregulated Collagen I,Collagen III,and FAP expressions,while miR-30 family expression was downregulated with miR-30b-5p being the most significant.Pue intervention inhibited the excessive proliferation of TGF-β1-induced hOMF,downregulated FAP,collagen type 3(COL3A1),collagen type 1(COL1A1),matrix metalloproteinase 1(MMP1),and matrix metalloproteinase 3(MMP3)expressions,and restored miR-30 family expression.Bioinformatics prediction and dual-luciferase reporter assay revealed that miR-30b-5p selectively inhibited FAP expression.Mechanistically,miR-30b-5p mimic suppressed the excessive proliferation of TGF-β1-induced hOMF and declinedfibrosis levels.Pue intervention significantly reversed the promotion of TGF-β1-induced OSF by miR-30b-5p inhibition.Conclusion:Pue mediated miR-30b-5p targeting FAP against OSF,which provided a theoretical basis for the pathogenesis research and Pue application in OSF.展开更多
Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be e...Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be effective in treating BLCA.The purpose of this study was to investigate the potential treating mechanisms of Puerarin on BLCA.Methods:The cell counting kit 8 assay and flow cytometry were performed to confirm Puerarin’s ability to suppress BLCA.The differentially expressed proteins(DEPs)were obtained by Tandem Mass Tags technology and functional enrichment analysis performed by R studio.The most enriched pathways were selected for study and the DEPs were screened out.Protein-protein interaction network maps were created using String and Cytoscape and key proteins,which will be analyzed for survival,expression,and upstream transcription factor prediction,were screened out using the cytoHubba plugin.CHEA3 was used to obtain upstream transcription factor validated by molecular docking and western blotting experiments.Results:Cell counting kit 8 showed that Puerarin inhibited BLCA cells,with 50%inhibitory concentration of 218μmol/L in T24 and 198μmol/L in 5637.Flow cytometry reveals that Puerarin blocks T24 and 5637 cells in G1 phase.1,385 DEPs were obtained and the enrichment analysis revealed that cell cycle and DNA replication were the two main areas in which DEPs were enriched.Cyclin-B-cyclin dependent kinase 1(CDK1),cyclin B1(CCNB1),and polo-like kinase 1(PLK1)were identified as key proteins,and their upstream transcription factor was predicted to be centromere protein A(CENPA).Puerarin’s binding energy to CENPA was determined by molecular docking to be−6.3 kcal/mol,indicating a strong binding interaction.Western blot showed that Puerarin significantly reduced the expression of CENPA.Conclusion:We hypothesize that Puerarin may inhibit the proliferation of bladder cancer cells by inhibiting CENPA expression to regulate PLK1 and CCNB1 expression,thereby affecting cell cycle.展开更多
Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three g...Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three groups(n=12).Twelve rats as the control group and the remaining rats were to establish a chronic periodontitis model ligated with orthodontic ligature wire on the cervical part of the left maxillary first molar under general anesthesia.All three groups were administered by gavage for 14 days:equal amounts of saline were given to the control and periodontitis groups,and 400 mg/kg concentration of puerarin was given to the puerarin group.All rats were anesthetized after 24 h of the last administration,blood was collected from the abdominal aorta,and the serum was centrifuged for the detection of IL-4,IL-10,IL-6,and IFN-γin peripheral blood,then all rats were executed,some rats separated from the left maxilla,fresh gingival tissue removed from the buccal-palatal side of the left maxillary first molar,and the remaining maxillary bone tissue used for the detection of Micro-CT;some rats subjected to the left maxillary bone specimens taken with the gingival tissues,used for HE staining detection.Results:HE staining showed that inflammatory cell infiltration was significantly reduced in the puerarin group compared to the periodontitis group.ELISA analysis showed that puerarin promoted IL-4 and IL-10 expression and decreased IL-6 and IFN-γexpression levels in serum(P<0.05).Micro-CT showed that puerarin significantly reduced alveolar bone resorption compared to the periodontitis group(P<0.05).Conclusion:Puerarin may inhibit inflammation and alveolar bone resorption in experimental periodontitis rats.展开更多
目的通过Meta分析评价葛根素对大鼠和小鼠骨密度的影响。方法通过中国知网、中国生物医学文献、万方、维普、PubMed、EMBase、Web of Science、Cochrane图书馆、Scopus数据库中自建库至2023年11月6日收录的有关葛根素治疗对大鼠和小鼠...目的通过Meta分析评价葛根素对大鼠和小鼠骨密度的影响。方法通过中国知网、中国生物医学文献、万方、维普、PubMed、EMBase、Web of Science、Cochrane图书馆、Scopus数据库中自建库至2023年11月6日收录的有关葛根素治疗对大鼠和小鼠骨密度影响的文献。文献纳入标准包括研究类型为随机对照试验(对照为安慰剂或空白组),研究对象为大鼠或小鼠,干预措施为葛根素,结果包含骨密度检测。文献排除标准包括葛根素联合其他药物治疗,没有原始研究数据,未公开发表,骨密度检测部位为下颌骨。采用SYRCLE'sRoB工具对纳入文献中的研究进行风险偏倚评估,采用Stata 16.0和Rev Man 5.3软件进行Meta分析。结果经数据库检索共获得429篇文献,根据纳入及排除标准最终纳入42篇文献。纳入文献中涉及41项研究,共925只动物纳入数据分析。与对照组相比,葛根素可改善大鼠和小鼠的骨密度:股骨37项研究,n=824,标准化均数差(standardized mean difference,SMD)=2.12,95%置信区间(confidence interval,CI)=1.69~2.54,P<0.0001、腰椎(13项研究,n=271,SMD=2.25,95%CI=1.49~3.01,P<0.0001)、胫骨(4项研究,n=95,SMD=0.94,95%CI=0.05~1.83,P=0.04)和全身(4项研究,n=94,SMD=1.89,95%CI=0.50~3.29,P=0.008)的组间骨密度差异均具有统计学意义。结论葛根素能够改善大鼠和小鼠的骨密度,本研究可以为葛根素防治骨质疏松症的临床研究提供良好的参考。展开更多
背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw2...背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw264.7分化为破骨细胞的影响。方法:将Raw264.7细胞分为7组干预培养,空白对照组采用DMEM高糖完全培养基培养,破骨细胞诱导组采用破骨诱导培养基培养,葛根素干预组在破骨诱导的同时加入50μmol/L葛根素培养,葛根素+Notch1 si RNA对照组、葛根素+Notch1 si RNA组、葛根素+Notch1过表达对照组、葛根素+Notch1过表达组分别采用Notch1 si RNA对照序列、Notch1 si RNA序列、Notch1过表达对照质粒、Notch1过表达质粒转染Raw264.7细胞后,加入破骨诱导培养基和葛根素进行培养。培养7 d后,采用抗酒石酸酸性磷酸酶染色观察破骨细胞的数量和大小,F-actin染色观察破骨细胞骨架形成情况,RT-PCR检测破骨细胞形成标志物的基因表达水平。结果与结论:(1)抗酒石酸酸性磷酸酶染色显示:葛根素干预可以抑制破骨细胞的生成,Notch1沉默会进一步减少破骨细胞的生成数量,Notch1过表达后破骨细胞生成数量明显增加;(2)F-actin染色显示:Raw264.7细胞经破骨诱导可以形成边界清晰的F-actin环,葛根素干预会抑制细胞骨架的形成,Notch1沉默会增强葛根素的抑制作用,而Notch1过表达则能减弱葛根素的抑制作用;(3)RT-PCR检测显示,葛根素可以抑制抗酒石酸酸性磷酸酶、组织蛋白酶K和c-Fos的m RNA表达,Notch1沉默后上述3个因子的m RNA表达进一步降低,Notch1过表达后上述3个因子的m RNA表达增加。结果表明:Notch信号通路在Raw264.7细胞分化为破骨细胞的过程中发挥作用,葛根素通过抑制Notch信号通路抑制Raw264.7细胞分化为破骨细胞。展开更多
葛根素是近年来中医药领域的主要研究热点之一。本研究利用文献计量学方法,梳理了PubMed和Web of science数据库近年发表的与葛根素相关的文献,通过关键词及热点突现分析,结合图谱内容解读,得出以下结论:葛根素在重大慢性疾病,如心脑血...葛根素是近年来中医药领域的主要研究热点之一。本研究利用文献计量学方法,梳理了PubMed和Web of science数据库近年发表的与葛根素相关的文献,通过关键词及热点突现分析,结合图谱内容解读,得出以下结论:葛根素在重大慢性疾病,如心脑血管缺血性疾病、代谢性疾病、肿瘤等的临床前模型中展现出显著的疗效,其主要作用机制与抗炎、抗氧化、抗凋亡等相关,但不同作用机制间的相互关系尚不完全清楚;葛根素在糖尿病肾病、神经退行性疾病以及肠道菌群紊乱等疾病的治疗中展现出有较高的潜力,相关研究值得进一步探索;葛根素联合用药的协同作用愈发被关注,但其生物利用度低、高质量临床试验缺乏等问题日益凸显。总的来说,葛根素具有巨大的潜在应用价值,但相关作用机理及其关系仍需深入挖掘,相关的高质量临床研究亦值得关注和重视。展开更多
基金This work was supported by the National Natural Science Foundation of China(Nos.81874496,82374530)the Clinical Medical Technology Innovation Guide Project of Hunan Province(No.2020SK53206)+3 种基金the Natural Science Foundation of Hunan Province(No.2021JJ70062)the Changsha Natural Science Foundation Project(No.kq2014019)the Health Special Fund Research Project of Hunan Province(No.B2020-07)the Clinical Pharmaceutical Research Fund of Hunan Medical Association(No.B202012).
文摘Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human oral mucosafibroblasts(hOMF)were induced with transforming growth factor beta1(TGF-β1)and intervened with Pue.Expressions offibrosis-related markers were analyzed by Western blot and IF staining.Cell viability was characterized by the CCK-8 assay.Expressions of miR-30 family members were quantified by qRT-PCR.The correlation betweenfibroblast activation protein(FAP)and miR-30 family expression was evaluated by the Pearson correlation coefficient.Bioinformatics prediction and dual-luciferase reporter assay were employed to verify the regulation between FAP and miR-30b-5p.The specific mechanism of Pue on OSF was explored through the promotion or inhibition of miR-30b-5p.Results:After induction by TGF-β1,hOMF showed upregulated Collagen I,Collagen III,and FAP expressions,while miR-30 family expression was downregulated with miR-30b-5p being the most significant.Pue intervention inhibited the excessive proliferation of TGF-β1-induced hOMF,downregulated FAP,collagen type 3(COL3A1),collagen type 1(COL1A1),matrix metalloproteinase 1(MMP1),and matrix metalloproteinase 3(MMP3)expressions,and restored miR-30 family expression.Bioinformatics prediction and dual-luciferase reporter assay revealed that miR-30b-5p selectively inhibited FAP expression.Mechanistically,miR-30b-5p mimic suppressed the excessive proliferation of TGF-β1-induced hOMF and declinedfibrosis levels.Pue intervention significantly reversed the promotion of TGF-β1-induced OSF by miR-30b-5p inhibition.Conclusion:Pue mediated miR-30b-5p targeting FAP against OSF,which provided a theoretical basis for the pathogenesis research and Pue application in OSF.
基金supported by National Natural Science Fund Item Number(82004110)Xuzhou Science and Technology Plan Project(KC22096)+3 种基金China Postdoctoral Science Foundation(2022M722674)Xuzhou Medical Reserve Talents Project(XWRCHT20220009)the Xuzhou Clinical Medicine Expert Team Project(2018TD004)Peixian Science and Technology Program(P202410)。
文摘Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be effective in treating BLCA.The purpose of this study was to investigate the potential treating mechanisms of Puerarin on BLCA.Methods:The cell counting kit 8 assay and flow cytometry were performed to confirm Puerarin’s ability to suppress BLCA.The differentially expressed proteins(DEPs)were obtained by Tandem Mass Tags technology and functional enrichment analysis performed by R studio.The most enriched pathways were selected for study and the DEPs were screened out.Protein-protein interaction network maps were created using String and Cytoscape and key proteins,which will be analyzed for survival,expression,and upstream transcription factor prediction,were screened out using the cytoHubba plugin.CHEA3 was used to obtain upstream transcription factor validated by molecular docking and western blotting experiments.Results:Cell counting kit 8 showed that Puerarin inhibited BLCA cells,with 50%inhibitory concentration of 218μmol/L in T24 and 198μmol/L in 5637.Flow cytometry reveals that Puerarin blocks T24 and 5637 cells in G1 phase.1,385 DEPs were obtained and the enrichment analysis revealed that cell cycle and DNA replication were the two main areas in which DEPs were enriched.Cyclin-B-cyclin dependent kinase 1(CDK1),cyclin B1(CCNB1),and polo-like kinase 1(PLK1)were identified as key proteins,and their upstream transcription factor was predicted to be centromere protein A(CENPA).Puerarin’s binding energy to CENPA was determined by molecular docking to be−6.3 kcal/mol,indicating a strong binding interaction.Western blot showed that Puerarin significantly reduced the expression of CENPA.Conclusion:We hypothesize that Puerarin may inhibit the proliferation of bladder cancer cells by inhibiting CENPA expression to regulate PLK1 and CCNB1 expression,thereby affecting cell cycle.
基金Hainan Provincial Financial Science and Technology Program Funded Project-Hainan Provincial Key Research and Development Social Development Project 2020(No.ZDYF2020166)Hainan Provincial Financial Science and Technology Program funded project-Hainan Provincial Key Research and Development Social Development Project 2023(No.ZDYF2023SHFZ095)。
文摘Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three groups(n=12).Twelve rats as the control group and the remaining rats were to establish a chronic periodontitis model ligated with orthodontic ligature wire on the cervical part of the left maxillary first molar under general anesthesia.All three groups were administered by gavage for 14 days:equal amounts of saline were given to the control and periodontitis groups,and 400 mg/kg concentration of puerarin was given to the puerarin group.All rats were anesthetized after 24 h of the last administration,blood was collected from the abdominal aorta,and the serum was centrifuged for the detection of IL-4,IL-10,IL-6,and IFN-γin peripheral blood,then all rats were executed,some rats separated from the left maxilla,fresh gingival tissue removed from the buccal-palatal side of the left maxillary first molar,and the remaining maxillary bone tissue used for the detection of Micro-CT;some rats subjected to the left maxillary bone specimens taken with the gingival tissues,used for HE staining detection.Results:HE staining showed that inflammatory cell infiltration was significantly reduced in the puerarin group compared to the periodontitis group.ELISA analysis showed that puerarin promoted IL-4 and IL-10 expression and decreased IL-6 and IFN-γexpression levels in serum(P<0.05).Micro-CT showed that puerarin significantly reduced alveolar bone resorption compared to the periodontitis group(P<0.05).Conclusion:Puerarin may inhibit inflammation and alveolar bone resorption in experimental periodontitis rats.
文摘目的通过Meta分析评价葛根素对大鼠和小鼠骨密度的影响。方法通过中国知网、中国生物医学文献、万方、维普、PubMed、EMBase、Web of Science、Cochrane图书馆、Scopus数据库中自建库至2023年11月6日收录的有关葛根素治疗对大鼠和小鼠骨密度影响的文献。文献纳入标准包括研究类型为随机对照试验(对照为安慰剂或空白组),研究对象为大鼠或小鼠,干预措施为葛根素,结果包含骨密度检测。文献排除标准包括葛根素联合其他药物治疗,没有原始研究数据,未公开发表,骨密度检测部位为下颌骨。采用SYRCLE'sRoB工具对纳入文献中的研究进行风险偏倚评估,采用Stata 16.0和Rev Man 5.3软件进行Meta分析。结果经数据库检索共获得429篇文献,根据纳入及排除标准最终纳入42篇文献。纳入文献中涉及41项研究,共925只动物纳入数据分析。与对照组相比,葛根素可改善大鼠和小鼠的骨密度:股骨37项研究,n=824,标准化均数差(standardized mean difference,SMD)=2.12,95%置信区间(confidence interval,CI)=1.69~2.54,P<0.0001、腰椎(13项研究,n=271,SMD=2.25,95%CI=1.49~3.01,P<0.0001)、胫骨(4项研究,n=95,SMD=0.94,95%CI=0.05~1.83,P=0.04)和全身(4项研究,n=94,SMD=1.89,95%CI=0.50~3.29,P=0.008)的组间骨密度差异均具有统计学意义。结论葛根素能够改善大鼠和小鼠的骨密度,本研究可以为葛根素防治骨质疏松症的临床研究提供良好的参考。
文摘背景:前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的:探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw264.7分化为破骨细胞的影响。方法:将Raw264.7细胞分为7组干预培养,空白对照组采用DMEM高糖完全培养基培养,破骨细胞诱导组采用破骨诱导培养基培养,葛根素干预组在破骨诱导的同时加入50μmol/L葛根素培养,葛根素+Notch1 si RNA对照组、葛根素+Notch1 si RNA组、葛根素+Notch1过表达对照组、葛根素+Notch1过表达组分别采用Notch1 si RNA对照序列、Notch1 si RNA序列、Notch1过表达对照质粒、Notch1过表达质粒转染Raw264.7细胞后,加入破骨诱导培养基和葛根素进行培养。培养7 d后,采用抗酒石酸酸性磷酸酶染色观察破骨细胞的数量和大小,F-actin染色观察破骨细胞骨架形成情况,RT-PCR检测破骨细胞形成标志物的基因表达水平。结果与结论:(1)抗酒石酸酸性磷酸酶染色显示:葛根素干预可以抑制破骨细胞的生成,Notch1沉默会进一步减少破骨细胞的生成数量,Notch1过表达后破骨细胞生成数量明显增加;(2)F-actin染色显示:Raw264.7细胞经破骨诱导可以形成边界清晰的F-actin环,葛根素干预会抑制细胞骨架的形成,Notch1沉默会增强葛根素的抑制作用,而Notch1过表达则能减弱葛根素的抑制作用;(3)RT-PCR检测显示,葛根素可以抑制抗酒石酸酸性磷酸酶、组织蛋白酶K和c-Fos的m RNA表达,Notch1沉默后上述3个因子的m RNA表达进一步降低,Notch1过表达后上述3个因子的m RNA表达增加。结果表明:Notch信号通路在Raw264.7细胞分化为破骨细胞的过程中发挥作用,葛根素通过抑制Notch信号通路抑制Raw264.7细胞分化为破骨细胞。
文摘葛根素是近年来中医药领域的主要研究热点之一。本研究利用文献计量学方法,梳理了PubMed和Web of science数据库近年发表的与葛根素相关的文献,通过关键词及热点突现分析,结合图谱内容解读,得出以下结论:葛根素在重大慢性疾病,如心脑血管缺血性疾病、代谢性疾病、肿瘤等的临床前模型中展现出显著的疗效,其主要作用机制与抗炎、抗氧化、抗凋亡等相关,但不同作用机制间的相互关系尚不完全清楚;葛根素在糖尿病肾病、神经退行性疾病以及肠道菌群紊乱等疾病的治疗中展现出有较高的潜力,相关研究值得进一步探索;葛根素联合用药的协同作用愈发被关注,但其生物利用度低、高质量临床试验缺乏等问题日益凸显。总的来说,葛根素具有巨大的潜在应用价值,但相关作用机理及其关系仍需深入挖掘,相关的高质量临床研究亦值得关注和重视。
