目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方...目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。展开更多
transcription and expression,and its potential biological mechanisms.Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended v...transcription and expression,and its potential biological mechanisms.Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended validation.Hg-exposed cell models and PTEN lowexpression models were established in vitro using 293T cells.PTEN gene expression was assessed using qRT-PCR,and Western blotting was used to measure PTEN,AKT,and PI3K protein levels.IL-6 expression was determined by ELISA.Results Combined findings from gene expression microarray analysis,bioinformatics,and population expansion validation indicated significant downregulation of the PTEN gene in the high-concentration Hg exposure group.In the Hg-exposed cell model(25 and 10μmol/L),a significant decrease in PTEN expression was observed,accompanied by a significant increase in PI3K,AKT,and IL-6 expression.Similarly,a low-expression cell model demonstrated that PTEN gene knockdown led to a significant decrease in PTEN protein expression and a substantial increase in PI3K,AKT,and IL-6 levels.Conclusion This is the first study to report that Hg exposure downregulates the PTEN gene,activates the PI3K/AKT regulatory pathway,and increases the expression of inflammatory factors,ultimately resulting in kidney inflammation.展开更多
Restenosis is a common complication following coronary angioplasty.The traditional use of seaweeds for health benefits has increasingly been explored,however few studies exist reporting its protective effects on the d...Restenosis is a common complication following coronary angioplasty.The traditional use of seaweeds for health benefits has increasingly been explored,however few studies exist reporting its protective effects on the development of restenosis and gut dysbiosis.The aim of this study was to investigate the potential of seaweed extracts(SE) of Ascophyllum nodosum and Fucus vesiculosus in inhibiting intimal hyperplasia in a rat model of restenosis and its underlying mechanisms in macrophages and vascular smooth muscle cells(vSMCs).16S rRNA sequencing was done to investigate the regulatory effect of SE on the gut microbiome of injured rats.As indicated by the results,SE significantly inhibited the progression of intimal hyperplasia in vivo,attenuated inflammation in macrophages and could inhibit the proliferation,dedifferentiation and migration of vSMCs.It was observed through immunoblotting assays that treatment with SE significantly upregulated PTEN expression in macrophages and inhibited the upregulation of PI3K and AKT expression in vSMCs.Meanwhile,according to the 16S rRNA gene sequencing analysis,supplementation with SE modulated gut microbiota composition in injured rats.In conclusion,SE could ameliorate intimal hyperplasia by inhibiting inflammation and vSMCs proliferation through the regulation of the PTEN/PI3K/AKT pathway and modulating the gut microbiome.展开更多
目的观察真武汤、越婢汤对阿霉素肾病大鼠肾组织磷酸酶张力蛋白基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)及肾脏自噬标志性分子微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)...目的观察真武汤、越婢汤对阿霉素肾病大鼠肾组织磷酸酶张力蛋白基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)及肾脏自噬标志性分子微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)表达的影响。方法将SD大鼠随机分为空白组、模型组、真武汤组(真武组)、越婢汤组(越婢组),每组12只。用阿霉素复制肾病综合征大鼠模型。造模成功后空白组和模型组以生理盐水灌胃,中药两组分别以真武汤和越婢汤颗粒剂灌胃,6周后检测血清白蛋白(ALB)、血肌酐(Scr)、尿素氮(BUN)及24 h尿蛋白定量(24-hour urine total protein quantity,24 h UTP)等,光镜观察肾脏病理,免疫组化观察大鼠肾组织PTEN、LC3的表达。结果与空白组相比,模型组肾脏病理可见部分肾小球肥大、毛细血管腔变窄,肾小管扩张、管腔可见蛋白质管型,间质可见炎性细胞浸润等;24 h UTP及Scr、BUN升高,ALB、LC3及PTEN表达明显降低,差异有统计学意义(P<0.05或P<0.01)。与模型组比较,中药两组肾脏病理改善,24 h UTP、Scr明显下降,ALB升高,肾组织LC3、PTEN表达增强,差异有统计学意义(P<0.05)。而真武组与越婢组比较无明显差异。结论真武汤和越婢汤均能改善阿霉素肾病大鼠的生化指标及肾脏病理,其作用机制可能与升高肾组织PTEN、LC3表达有关。展开更多
文摘目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。
基金supported by the Jiangsu Province’s Outstanding Medical Academic Leader Program[CXTDA2017029]and the Jiangsu Provincial Key Medical Discipline[ZDXK202249].
文摘transcription and expression,and its potential biological mechanisms.Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended validation.Hg-exposed cell models and PTEN lowexpression models were established in vitro using 293T cells.PTEN gene expression was assessed using qRT-PCR,and Western blotting was used to measure PTEN,AKT,and PI3K protein levels.IL-6 expression was determined by ELISA.Results Combined findings from gene expression microarray analysis,bioinformatics,and population expansion validation indicated significant downregulation of the PTEN gene in the high-concentration Hg exposure group.In the Hg-exposed cell model(25 and 10μmol/L),a significant decrease in PTEN expression was observed,accompanied by a significant increase in PI3K,AKT,and IL-6 expression.Similarly,a low-expression cell model demonstrated that PTEN gene knockdown led to a significant decrease in PTEN protein expression and a substantial increase in PI3K,AKT,and IL-6 levels.Conclusion This is the first study to report that Hg exposure downregulates the PTEN gene,activates the PI3K/AKT regulatory pathway,and increases the expression of inflammatory factors,ultimately resulting in kidney inflammation.
基金supported by the research grants from the Ministry of Science and Technology of Taiwan (MOST108-2320B-038-040-MY3 and MOST 111-2320-B-038-049)the National Science and Technology Council (NSTC111-2320-B-038-049)。
文摘Restenosis is a common complication following coronary angioplasty.The traditional use of seaweeds for health benefits has increasingly been explored,however few studies exist reporting its protective effects on the development of restenosis and gut dysbiosis.The aim of this study was to investigate the potential of seaweed extracts(SE) of Ascophyllum nodosum and Fucus vesiculosus in inhibiting intimal hyperplasia in a rat model of restenosis and its underlying mechanisms in macrophages and vascular smooth muscle cells(vSMCs).16S rRNA sequencing was done to investigate the regulatory effect of SE on the gut microbiome of injured rats.As indicated by the results,SE significantly inhibited the progression of intimal hyperplasia in vivo,attenuated inflammation in macrophages and could inhibit the proliferation,dedifferentiation and migration of vSMCs.It was observed through immunoblotting assays that treatment with SE significantly upregulated PTEN expression in macrophages and inhibited the upregulation of PI3K and AKT expression in vSMCs.Meanwhile,according to the 16S rRNA gene sequencing analysis,supplementation with SE modulated gut microbiota composition in injured rats.In conclusion,SE could ameliorate intimal hyperplasia by inhibiting inflammation and vSMCs proliferation through the regulation of the PTEN/PI3K/AKT pathway and modulating the gut microbiome.
文摘目的观察真武汤、越婢汤对阿霉素肾病大鼠肾组织磷酸酶张力蛋白基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)及肾脏自噬标志性分子微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)表达的影响。方法将SD大鼠随机分为空白组、模型组、真武汤组(真武组)、越婢汤组(越婢组),每组12只。用阿霉素复制肾病综合征大鼠模型。造模成功后空白组和模型组以生理盐水灌胃,中药两组分别以真武汤和越婢汤颗粒剂灌胃,6周后检测血清白蛋白(ALB)、血肌酐(Scr)、尿素氮(BUN)及24 h尿蛋白定量(24-hour urine total protein quantity,24 h UTP)等,光镜观察肾脏病理,免疫组化观察大鼠肾组织PTEN、LC3的表达。结果与空白组相比,模型组肾脏病理可见部分肾小球肥大、毛细血管腔变窄,肾小管扩张、管腔可见蛋白质管型,间质可见炎性细胞浸润等;24 h UTP及Scr、BUN升高,ALB、LC3及PTEN表达明显降低,差异有统计学意义(P<0.05或P<0.01)。与模型组比较,中药两组肾脏病理改善,24 h UTP、Scr明显下降,ALB升高,肾组织LC3、PTEN表达增强,差异有统计学意义(P<0.05)。而真武组与越婢组比较无明显差异。结论真武汤和越婢汤均能改善阿霉素肾病大鼠的生化指标及肾脏病理,其作用机制可能与升高肾组织PTEN、LC3表达有关。
