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
MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to...MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to provide a theoretical basis for treatment.Bioinformatics was used to analyze pituitary adenoma-related genes and screen new targets related to RECK and miRNA.As well,the relationship between miR-200b-3p and RECK protein was verified using a double-luciferase reporter gene assay.The expression of miR-200b-3p in clinical samples was analyzed by in situ hybridization.Transfection of the miR-200b-3p inhibitor and small interfering-RECK(si-RECK)was verified by qPCR.GH3 cell viability and proliferation were detected using CCK8 and EdU assays.Apoptosis was detected by flow cytometry and western blotting.Wound healing and Transwell assays were used to detect cell migration and invasion.The effects of miR-200b-3p and RECK on GH3 cells were verified using salvage experiments.miR-200b-3p was highly expressed in pituitary tumor tissue.Inhibitors of miR-200b-3p inhibited cell proliferation promoted cell apoptosis,inhibited invasion and migration,and inhibited the expression of matrix metalloproteinases.Interestingly,miR-200b-3p negatively regulated RECK.The expression of RECK in pituitary adenoma tissues was lower than that in neighboring tissues.Si-RECK rescued the function of miR-200b-3p inhibitors in the above cellular behaviors,and miR-200b-3p accelerated the development of pituitary adenoma by negatively regulating RECK expression.In summary,this study investigated the molecular mechanism by which miR-200b-3p regulates the progression of pituitary adenoma through the negative regulation of RECK.The findings provide a new target for the treatment of pituitary adenoma.展开更多
目的探讨葛根素调控miR-30b-5p表达对类固醇诱导的兔股骨头坏死(osteonecrosis of the femoral head,ONFH)的作用及机制。方法将84只新西兰大白兔分为对照组、ONFH组、葛根素低剂量组、葛根素高剂量组、阳性对照组、葛根素高剂量+antago...目的探讨葛根素调控miR-30b-5p表达对类固醇诱导的兔股骨头坏死(osteonecrosis of the femoral head,ONFH)的作用及机制。方法将84只新西兰大白兔分为对照组、ONFH组、葛根素低剂量组、葛根素高剂量组、阳性对照组、葛根素高剂量+antagomir NC组、葛根素高剂量+miR-30b-5p antagomir组,每组12只。通过Micro-CT检测大白兔股骨头骨小梁数量、骨小梁厚度及骨小梁分离度;HE染色检测股骨头组织病理变化;ELISA法检测股骨头组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;TUNEL染色检测股骨头组织中骨细胞凋亡;qRT-PCR检测股骨头组织中miR-30b-5p表达;Western Blot检测股骨头组织中Dickkopf-1(DKK-1)蛋白表达。结果与对照组比较,ONFH组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达降低,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达升高(P<0.05);与ONFH组比较,葛根素低、高剂量组、阳性对照组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达升高,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达降低(P<0.05);miR-30b-5p antagomir减弱了高剂量葛根素对ONFH兔氧化应激及细胞凋亡的抑制作用。结论葛根素可抑制氧化应激及细胞凋亡进而发挥对ONFH兔的保护作用,其机制可能与上调miR-30b-5p表达有关。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 Correlation between RECK and GH-type pituitary adenomas(No.21JR11RE027).
文摘MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to provide a theoretical basis for treatment.Bioinformatics was used to analyze pituitary adenoma-related genes and screen new targets related to RECK and miRNA.As well,the relationship between miR-200b-3p and RECK protein was verified using a double-luciferase reporter gene assay.The expression of miR-200b-3p in clinical samples was analyzed by in situ hybridization.Transfection of the miR-200b-3p inhibitor and small interfering-RECK(si-RECK)was verified by qPCR.GH3 cell viability and proliferation were detected using CCK8 and EdU assays.Apoptosis was detected by flow cytometry and western blotting.Wound healing and Transwell assays were used to detect cell migration and invasion.The effects of miR-200b-3p and RECK on GH3 cells were verified using salvage experiments.miR-200b-3p was highly expressed in pituitary tumor tissue.Inhibitors of miR-200b-3p inhibited cell proliferation promoted cell apoptosis,inhibited invasion and migration,and inhibited the expression of matrix metalloproteinases.Interestingly,miR-200b-3p negatively regulated RECK.The expression of RECK in pituitary adenoma tissues was lower than that in neighboring tissues.Si-RECK rescued the function of miR-200b-3p inhibitors in the above cellular behaviors,and miR-200b-3p accelerated the development of pituitary adenoma by negatively regulating RECK expression.In summary,this study investigated the molecular mechanism by which miR-200b-3p regulates the progression of pituitary adenoma through the negative regulation of RECK.The findings provide a new target for the treatment of pituitary adenoma.
文摘目的探讨葛根素调控miR-30b-5p表达对类固醇诱导的兔股骨头坏死(osteonecrosis of the femoral head,ONFH)的作用及机制。方法将84只新西兰大白兔分为对照组、ONFH组、葛根素低剂量组、葛根素高剂量组、阳性对照组、葛根素高剂量+antagomir NC组、葛根素高剂量+miR-30b-5p antagomir组,每组12只。通过Micro-CT检测大白兔股骨头骨小梁数量、骨小梁厚度及骨小梁分离度;HE染色检测股骨头组织病理变化;ELISA法检测股骨头组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;TUNEL染色检测股骨头组织中骨细胞凋亡;qRT-PCR检测股骨头组织中miR-30b-5p表达;Western Blot检测股骨头组织中Dickkopf-1(DKK-1)蛋白表达。结果与对照组比较,ONFH组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达降低,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达升高(P<0.05);与ONFH组比较,葛根素低、高剂量组、阳性对照组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达升高,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达降低(P<0.05);miR-30b-5p antagomir减弱了高剂量葛根素对ONFH兔氧化应激及细胞凋亡的抑制作用。结论葛根素可抑制氧化应激及细胞凋亡进而发挥对ONFH兔的保护作用,其机制可能与上调miR-30b-5p表达有关。
