BACKGROUND In China banxia xiexin decoction(BXD)has been used in treating gastric cancer(GC)for thousands of years and BXD has a good role in reversing GC histopathology,but its chemical composition and action mechani...BACKGROUND In China banxia xiexin decoction(BXD)has been used in treating gastric cancer(GC)for thousands of years and BXD has a good role in reversing GC histopathology,but its chemical composition and action mechanism are still unknown.AIM To investigate the mechanism of action of BXD against GC based on transcriptomics,network pharmacology,in vivo and in vitro experiments.METHODS The transplanted tumor model was prepared,and the nude mouse were pathologically examined after administration,and hematoxylin-eosin staining was performed.The active ingredients of BXD were quality controlled and identified using ultra-performance liquid chromatography tandem quadrupole electrostatic field orbitrap mass spectrometry(UPLC-Q-Orbitrap MS/MS),and traditional Chinese medicines systems pharmacology platform,drug bank and the Swiss target prediction platform to predict the relevant targets,the differentially expressed genes(DEGs)of GC were screened by RNA-seq sequencing,and the overlapping targets were analyzed to obtain the key targets and pathways.Cell Counting Kit-8,apoptosis assay,cell migration and Realtime fluorescence quantitative polymerase chain reaction were used for in vitro experiments.RESULTS All dosing groups inhibited the growth of transplanted tumors in laboratory-bred strain nude,with the capecitabine group and the BXD medium-dose group being the best.A total of 29 compounds and 859 potential targets in BXD were identified by UPLC-Q-Orbitrap MS/MS and network pharmacology,RNA-seq sequencing found 4767 GC DEGs,which were combined with network pharmacology and analyzed 246 potential therapeutic targets were obtained and pathway results showed that BXD may against GC through the Phosphoinositide 3-kinase(PI3K)/protein kinase B(AKt)signaling pathway.In vitro cellular experiments confirmed that BXDcontaining serum and LY294002 could inhibit the proliferation of GC cells,promote apoptosis,and inhibit the migration of GC cells by decreasing the expression of EGFR,PIK3CA,IL6,BCL2 and AKT1 in the PI3K-Akt pathway in MGC-803 expression.CONCLUSION BXD has the effect of inhibiting tumor growth rate and delaying the development of GC.Its mechanism of action may be related to the regulation of PI3K-Akt signaling pathway.展开更多
Objective: To investigate the differential expression of miRNA and related biological functions and signaling pathways after the intervention of the THP-1-derived foam cell model with the drug-containing serum of Yima...Objective: To investigate the differential expression of miRNA and related biological functions and signaling pathways after the intervention of the THP-1-derived foam cell model with the drug-containing serum of Yimaijiangzhi Decoction. Methods: The experiment was divided into macrophage group, foam cell model group and Yimaijiangzhi drug-containing serum group. THP-1 cells were induced into macrophages by Fopol ester, and induced differentiated macrophages were given ox-LDL to establish foam cell model, and Yimaijiangzhi decoction rat serum was used to intervene the foam cells. Total RNA was extracted from cells in each group for miRNA sequencing, differential expression of miRNA was screened, and relevant target genes were predicted for GO analysis and KEGG analysis, protein interaction network and miRNA-target gene interaction network were established, and RT-qPCR was used to verify the possible signaling pathways for improving atherosclerosis. Result: The difference miRNA between blank group and model group was hsa-miR-302c-3p, hsa-miR-302d-3p, hsa- mir-30d-3p, hsa-mir-3189-3p, hsa-mir-374b-5p, hsa-mir-423-5p, hsa-mir-423-5p, and hsa- mir-4781-3p, hsa-mir-663a;The miRNAs of model group and Yimaijiangzhi drug-containing serum group were hsa-mir-3150a-3p, hsa-mir-7704, hsa-mir-887-3p, hsa-mir-150-5p, hsa- mir-423-5p, hsa-mir-374c-3p, hsa-mir-374c-3p, hsa-mir-374b-5p;The difference of miRNAs prediction target genes between model group and Yimaijiangzhi drug-containing serum group showed that the miRNA prediction target genes were mainly enriched in MAPK signaling pathway, ErbB signaling pathway, Hippo signaling pathway, Wnt signaling pathway and other signaling pathways. SCN1A, PRKACA, MECP2, EIF4E, SRSF1, MBNL1, PRKCA, PPARGC1A may be the potential key targets for the effect of the drug-containing serum of Yimaijiangzhi Decoction on THP-1-derived foam cells. Conclusion: hsa-mir-374c-3p, hsa- mir-423-5p, and hsa-mir-374b-5p are important miRNAs that the drug-containing serum of Yimaijiangzhi Decoction acts on foam cells. The significantly differentially expressed mirnas and significantly enriched related signaling pathways may provide new ideas for the diagnosis and treatment of atherosclerosis.展开更多
Huatan Jiangzhuo decoction(HJD)is a combination of six traditional Chinese medicines that were used for lipid metabolism-related disorders,but its efficacy and underlying mechanisms have not been explored by modern re...Huatan Jiangzhuo decoction(HJD)is a combination of six traditional Chinese medicines that were used for lipid metabolism-related disorders,but its efficacy and underlying mechanisms have not been explored by modern research strategies.This study aimed to investigate the therapeutic role of HJD in determining the transcriptome level.Hyperlipidemia model was established by feeding Sprague–Dawley rats with high-fat diet.Differentially expressed genes(DEGs)were detected by high-through transcriptome sequencing,followed by gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.The total cholesterol(TC)and triglyceride(TG)levels in hyperlipidemia model rats were significantly increased,whereas high-density lipoprotein(HDL)concentration decreased when compared to normal rats,and HJD significantly downregulated TC concentrations and liver coefficient in the hyperlipidemia rats.Histology staining showed that HDJ greatly recovered the lipid accumulation in rat hepatic stellate cells and aortic arch vascular wall thickness of hyperlipidemia rats.One thousand nine hundred and thirty-six DEGs were identified in the HJD-treated hyperlipidemia rats,which were associated with various biological processes and signaling pathways such as peroxisome proliferator-activated receptors,AMP-activated Protein Kinase,and insulin signaling pathways.Quantitative reverse transcription–polymerase chain reaction further confirmed the downregulated expression of cholesterol 7-α-hydroxylase(CYP7A1),liver orphan receptor(LXRα),peroxisome proliferator-activated receptor gamma(PPARγ),and Sterol Response Element-Binding Protein 1c(SREBP1c)genes in hyperlipidemia rats treated with HJD.Our data first elucidated the gene expression profile of high-fat diet-induced hyperlipidemia in rats after HJD treatment,and lipid metabolism-related genes(CYP7A1,LXRα,PPARγ,and SREBP1c)may be potentially biomarkers for HJD-alleviated hyperlipidemia.展开更多
目的:采用高通量测序技术探讨六味地黄汤对肝肾阴虚型妊娠期便秘患者肠道菌群的影响。方法:选取肝肾阴虚型妊娠期便秘患者28例,分别于治疗前和服用六味地黄汤2周后收集其粪便标本各50 g,采用QIAamp DNA Stool Mini Kit试剂盒提取治疗前...目的:采用高通量测序技术探讨六味地黄汤对肝肾阴虚型妊娠期便秘患者肠道菌群的影响。方法:选取肝肾阴虚型妊娠期便秘患者28例,分别于治疗前和服用六味地黄汤2周后收集其粪便标本各50 g,采用QIAamp DNA Stool Mini Kit试剂盒提取治疗前后粪便样本的总基因组脱氧核糖核酸(DNA),通过聚合酶链反应(PCR)扩增获得16S rRNA基因V4标签片段,构建Illumina平台文库并测序;采用定量分析微生物生态学(QIIME)软件分析肠道菌群变化,绘制稀释曲线,分析治疗前后粪便样本生物信息学,进行Alpha多样性指数分析,并分别使用Chao1、Shannon及Simpson指数计算肠道菌群丰度和菌群多样性;采用LefSe技术分析治疗前后粪便样本中菌群组成中的差异性菌属。结果:稀释曲线表明测序数据量合理,基本能覆盖样本中所有的细菌种类;六味地黄汤治疗后患者肠道菌群的Chao1指数高于治疗前(P<0.05),但Shannon指数和Simpson指数在治疗前后比较,差异均无统计学意义(P>0.05);治疗前后患者肠道菌群在门水平中均以bacteroidetes、firmicutes和proteobacteria为优势菌群,治疗后在属水平肠道菌群共有10个菌属丰度发生明显改变,其中escherichia、alistipes、parabacteroides、bacteroides fragilis、coprococcus菌属丰度较治疗前明显降低,faecalibacterium、roseburia、bifidobacterium、prevotella、lactobacillus菌属丰度较治疗前升高(P<0.05)。结论:妊娠期便秘患者服用六味地黄汤后,肠道内菌群结构改变显著,尤其在属水平有明显差异。展开更多
目的:探讨活血降糖饮治疗糖尿病肾病的分子机制。方法:通过高脂饲料喂养加尾静脉注射链脉佐菌素(STZ)等建立糖尿病肾病大鼠模型,并将模型大鼠随机分为糖尿病肾病(DN)组、模型+双胍+中药组(DN+Met+ZY)、模型+双胍+卡托普利组(DN+Met+CP)...目的:探讨活血降糖饮治疗糖尿病肾病的分子机制。方法:通过高脂饲料喂养加尾静脉注射链脉佐菌素(STZ)等建立糖尿病肾病大鼠模型,并将模型大鼠随机分为糖尿病肾病(DN)组、模型+双胍+中药组(DN+Met+ZY)、模型+双胍+卡托普利组(DN+Met+CP),给予相应药物进行灌胃治疗,并另设正常对照组,每组10只,干预16周后,观察各组大鼠空腹血糖(FBG)、血脂、血清尿素氮(BUN)、血肌酐(Scr)、24 h尿总蛋白(24 h UTP)及尿微量清蛋白(24 h mALB)的变化。观察肾组织病理改变,并对各组肾组织进行RNA-seq测序。结果:与模型组比较,中药组和卡托普利组可以显著下调糖尿病肾病大鼠FBG、TC、TG、LDL-C、Scr、BUN、24 h UTP、24 h mALB等水平,而HDL-C无明显变化;与卡托普利组之间比较,中药组FBG、TC、TG、LDL-C明显下降,HDL-c、Scr、BUN、24 h UTP、24 h mALB均无明显差别。与模型组比较,中药组和卡托普利组均可以显著改善糖尿病肾病大鼠的肾脏病理学变化。转录组测序结果显示模型组对比正常组与中药组对比模型组转录本中基因总量接近,其中表达差异最明显的双向调控基因共筛选出10个;京都基因与基因组百科全书(KEGG)前20条显著富集通路分析显示模型组对比正常组与中药组对比模型组共同作用的信号通路有代谢相关通路及cAMP信号通路。结论:活血降糖饮联合二甲双胍能改善糖尿病肾病大鼠的糖脂代谢紊乱,减少尿白蛋白的排泄,改善肾功能和肾脏病理改变,延缓糖尿病肾病病情进展,其作用机制可能是通过Cftr、Pdk4、Angptl4、Hmgcs2等多靶点介导多通路实现的。展开更多
基金Supported by the Key Program of Shandong Province,China,No.2016CYJS08A01-6.
文摘BACKGROUND In China banxia xiexin decoction(BXD)has been used in treating gastric cancer(GC)for thousands of years and BXD has a good role in reversing GC histopathology,but its chemical composition and action mechanism are still unknown.AIM To investigate the mechanism of action of BXD against GC based on transcriptomics,network pharmacology,in vivo and in vitro experiments.METHODS The transplanted tumor model was prepared,and the nude mouse were pathologically examined after administration,and hematoxylin-eosin staining was performed.The active ingredients of BXD were quality controlled and identified using ultra-performance liquid chromatography tandem quadrupole electrostatic field orbitrap mass spectrometry(UPLC-Q-Orbitrap MS/MS),and traditional Chinese medicines systems pharmacology platform,drug bank and the Swiss target prediction platform to predict the relevant targets,the differentially expressed genes(DEGs)of GC were screened by RNA-seq sequencing,and the overlapping targets were analyzed to obtain the key targets and pathways.Cell Counting Kit-8,apoptosis assay,cell migration and Realtime fluorescence quantitative polymerase chain reaction were used for in vitro experiments.RESULTS All dosing groups inhibited the growth of transplanted tumors in laboratory-bred strain nude,with the capecitabine group and the BXD medium-dose group being the best.A total of 29 compounds and 859 potential targets in BXD were identified by UPLC-Q-Orbitrap MS/MS and network pharmacology,RNA-seq sequencing found 4767 GC DEGs,which were combined with network pharmacology and analyzed 246 potential therapeutic targets were obtained and pathway results showed that BXD may against GC through the Phosphoinositide 3-kinase(PI3K)/protein kinase B(AKt)signaling pathway.In vitro cellular experiments confirmed that BXDcontaining serum and LY294002 could inhibit the proliferation of GC cells,promote apoptosis,and inhibit the migration of GC cells by decreasing the expression of EGFR,PIK3CA,IL6,BCL2 and AKT1 in the PI3K-Akt pathway in MGC-803 expression.CONCLUSION BXD has the effect of inhibiting tumor growth rate and delaying the development of GC.Its mechanism of action may be related to the regulation of PI3K-Akt signaling pathway.
基金Guangxi Natural Science Foundation(No.2020GXNSFAA297158)The Fifth Batch of National Clinical Excellent Talent Training Projects[Guozhong Pharmaceutical Education(2022)No.1]+1 种基金Guangxi Youth Qihuang Scholar Training Program[Guizhong Medical Science and Education Development(2022)No.13]Guangxi Graduate Education Innovation Program Project(No.YCSW2022340)。
文摘Objective: To investigate the differential expression of miRNA and related biological functions and signaling pathways after the intervention of the THP-1-derived foam cell model with the drug-containing serum of Yimaijiangzhi Decoction. Methods: The experiment was divided into macrophage group, foam cell model group and Yimaijiangzhi drug-containing serum group. THP-1 cells were induced into macrophages by Fopol ester, and induced differentiated macrophages were given ox-LDL to establish foam cell model, and Yimaijiangzhi decoction rat serum was used to intervene the foam cells. Total RNA was extracted from cells in each group for miRNA sequencing, differential expression of miRNA was screened, and relevant target genes were predicted for GO analysis and KEGG analysis, protein interaction network and miRNA-target gene interaction network were established, and RT-qPCR was used to verify the possible signaling pathways for improving atherosclerosis. Result: The difference miRNA between blank group and model group was hsa-miR-302c-3p, hsa-miR-302d-3p, hsa- mir-30d-3p, hsa-mir-3189-3p, hsa-mir-374b-5p, hsa-mir-423-5p, hsa-mir-423-5p, and hsa- mir-4781-3p, hsa-mir-663a;The miRNAs of model group and Yimaijiangzhi drug-containing serum group were hsa-mir-3150a-3p, hsa-mir-7704, hsa-mir-887-3p, hsa-mir-150-5p, hsa- mir-423-5p, hsa-mir-374c-3p, hsa-mir-374c-3p, hsa-mir-374b-5p;The difference of miRNAs prediction target genes between model group and Yimaijiangzhi drug-containing serum group showed that the miRNA prediction target genes were mainly enriched in MAPK signaling pathway, ErbB signaling pathway, Hippo signaling pathway, Wnt signaling pathway and other signaling pathways. SCN1A, PRKACA, MECP2, EIF4E, SRSF1, MBNL1, PRKCA, PPARGC1A may be the potential key targets for the effect of the drug-containing serum of Yimaijiangzhi Decoction on THP-1-derived foam cells. Conclusion: hsa-mir-374c-3p, hsa- mir-423-5p, and hsa-mir-374b-5p are important miRNAs that the drug-containing serum of Yimaijiangzhi Decoction acts on foam cells. The significantly differentially expressed mirnas and significantly enriched related signaling pathways may provide new ideas for the diagnosis and treatment of atherosclerosis.
基金the Natural Science Foundation of Guangdong Province,China(No.2020A1515010725).
文摘Huatan Jiangzhuo decoction(HJD)is a combination of six traditional Chinese medicines that were used for lipid metabolism-related disorders,but its efficacy and underlying mechanisms have not been explored by modern research strategies.This study aimed to investigate the therapeutic role of HJD in determining the transcriptome level.Hyperlipidemia model was established by feeding Sprague–Dawley rats with high-fat diet.Differentially expressed genes(DEGs)were detected by high-through transcriptome sequencing,followed by gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.The total cholesterol(TC)and triglyceride(TG)levels in hyperlipidemia model rats were significantly increased,whereas high-density lipoprotein(HDL)concentration decreased when compared to normal rats,and HJD significantly downregulated TC concentrations and liver coefficient in the hyperlipidemia rats.Histology staining showed that HDJ greatly recovered the lipid accumulation in rat hepatic stellate cells and aortic arch vascular wall thickness of hyperlipidemia rats.One thousand nine hundred and thirty-six DEGs were identified in the HJD-treated hyperlipidemia rats,which were associated with various biological processes and signaling pathways such as peroxisome proliferator-activated receptors,AMP-activated Protein Kinase,and insulin signaling pathways.Quantitative reverse transcription–polymerase chain reaction further confirmed the downregulated expression of cholesterol 7-α-hydroxylase(CYP7A1),liver orphan receptor(LXRα),peroxisome proliferator-activated receptor gamma(PPARγ),and Sterol Response Element-Binding Protein 1c(SREBP1c)genes in hyperlipidemia rats treated with HJD.Our data first elucidated the gene expression profile of high-fat diet-induced hyperlipidemia in rats after HJD treatment,and lipid metabolism-related genes(CYP7A1,LXRα,PPARγ,and SREBP1c)may be potentially biomarkers for HJD-alleviated hyperlipidemia.
文摘目的:采用高通量测序技术探讨六味地黄汤对肝肾阴虚型妊娠期便秘患者肠道菌群的影响。方法:选取肝肾阴虚型妊娠期便秘患者28例,分别于治疗前和服用六味地黄汤2周后收集其粪便标本各50 g,采用QIAamp DNA Stool Mini Kit试剂盒提取治疗前后粪便样本的总基因组脱氧核糖核酸(DNA),通过聚合酶链反应(PCR)扩增获得16S rRNA基因V4标签片段,构建Illumina平台文库并测序;采用定量分析微生物生态学(QIIME)软件分析肠道菌群变化,绘制稀释曲线,分析治疗前后粪便样本生物信息学,进行Alpha多样性指数分析,并分别使用Chao1、Shannon及Simpson指数计算肠道菌群丰度和菌群多样性;采用LefSe技术分析治疗前后粪便样本中菌群组成中的差异性菌属。结果:稀释曲线表明测序数据量合理,基本能覆盖样本中所有的细菌种类;六味地黄汤治疗后患者肠道菌群的Chao1指数高于治疗前(P<0.05),但Shannon指数和Simpson指数在治疗前后比较,差异均无统计学意义(P>0.05);治疗前后患者肠道菌群在门水平中均以bacteroidetes、firmicutes和proteobacteria为优势菌群,治疗后在属水平肠道菌群共有10个菌属丰度发生明显改变,其中escherichia、alistipes、parabacteroides、bacteroides fragilis、coprococcus菌属丰度较治疗前明显降低,faecalibacterium、roseburia、bifidobacterium、prevotella、lactobacillus菌属丰度较治疗前升高(P<0.05)。结论:妊娠期便秘患者服用六味地黄汤后,肠道内菌群结构改变显著,尤其在属水平有明显差异。
文摘目的:探讨活血降糖饮治疗糖尿病肾病的分子机制。方法:通过高脂饲料喂养加尾静脉注射链脉佐菌素(STZ)等建立糖尿病肾病大鼠模型,并将模型大鼠随机分为糖尿病肾病(DN)组、模型+双胍+中药组(DN+Met+ZY)、模型+双胍+卡托普利组(DN+Met+CP),给予相应药物进行灌胃治疗,并另设正常对照组,每组10只,干预16周后,观察各组大鼠空腹血糖(FBG)、血脂、血清尿素氮(BUN)、血肌酐(Scr)、24 h尿总蛋白(24 h UTP)及尿微量清蛋白(24 h mALB)的变化。观察肾组织病理改变,并对各组肾组织进行RNA-seq测序。结果:与模型组比较,中药组和卡托普利组可以显著下调糖尿病肾病大鼠FBG、TC、TG、LDL-C、Scr、BUN、24 h UTP、24 h mALB等水平,而HDL-C无明显变化;与卡托普利组之间比较,中药组FBG、TC、TG、LDL-C明显下降,HDL-c、Scr、BUN、24 h UTP、24 h mALB均无明显差别。与模型组比较,中药组和卡托普利组均可以显著改善糖尿病肾病大鼠的肾脏病理学变化。转录组测序结果显示模型组对比正常组与中药组对比模型组转录本中基因总量接近,其中表达差异最明显的双向调控基因共筛选出10个;京都基因与基因组百科全书(KEGG)前20条显著富集通路分析显示模型组对比正常组与中药组对比模型组共同作用的信号通路有代谢相关通路及cAMP信号通路。结论:活血降糖饮联合二甲双胍能改善糖尿病肾病大鼠的糖脂代谢紊乱,减少尿白蛋白的排泄,改善肾功能和肾脏病理改变,延缓糖尿病肾病病情进展,其作用机制可能是通过Cftr、Pdk4、Angptl4、Hmgcs2等多靶点介导多通路实现的。
