BACKGROUND Obesity and type 2 diabetes mellitus(T2DM)are frequent co-occurring disorders that affect regular metabolic functions.Obesity has also been linked to an inc-reased risk of developing diabetes.Obesity and di...BACKGROUND Obesity and type 2 diabetes mellitus(T2DM)are frequent co-occurring disorders that affect regular metabolic functions.Obesity has also been linked to an inc-reased risk of developing diabetes.Obesity and diabetes are on the rise,increa-sing healthcare costs and raising mortality rates.Research has revealed that the expression profile of microRNAs(miRNAs)changes as diabetes progresses.Fur-thermore,vitamin D may have an anti-obesity effect and inverse association with body weight and body mass index(BMI).Low vitamin D levels do not solely cause obesity,which could be a factor in the etiology of T2DM.METHODS This study included 210 participants,of which,82 were obese(BMI>30 kg/m2)without T2DM,28 were obese with T2DM,and 100 were healthy controls.BMI was evaluated and both fasting and postprandial blood glucose were used to confirm T2DM.Exosomal miRNA-200a and miRNA-200b expression were analyzed using real-time PCR using Taqman probes,and vitamin-D levels were evaluated using an electrochemiluminescence-based immunoassay technique.All data analyses were performed using SPSS 20.0 and GraphPad Prism 5 software.RESULTS Overall,a 2.20-and 4.40-fold increase in miRNA-200a and miRNA-200b expression was observed among participants compared to healthy controls.MiRNA-200a and miRNA-200b expression among obese participants increased 2.40-fold and 3.93-fold,respectively,while in obese T2DM participants these values were 2.67-fold,and 5.78-fold,respectively,and these differences were found to be statistically significant(P=0.02)(P<0.0001).Obese participants showed a vitamin D level of 34.27 ng/mL,while in obese-T2DM participants vitamin D level was 22.21 ng/mL(P<0.0001).Vitamin D was negatively correlated with miRNA-200a(r=-0.22,P=0.01)and miRNA-200b(r=-0.19,P=0.04).MiRNA-200a sensitivity was 75%,and specificity was 57%,with a cutoff value of 2.07-fold.MiRNA-200b sensitivity was 75%,and specificity was 71%with a cutoff value of 4.12-fold,suggesting that miRNA-200a and miRNA-200b with an increased expression of 2.07-and 4.12-fold could be predictive indicators for the risk of diabetes in obese participants.CONCLUSION MiRNA-200a and miRNA-200b were higher in diabetic obese participants vs non-diabetic obese participants,and insufficient vitamin D levels in obese T2DM participants may be involved in poor clinical outcome.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
Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance ...Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance and differentiation,inhibiting canonical autophagy via deletion of core genes,such as Atg5,Atg16l1,and Atg7,or blockade of canonical interactions between FIP200 and ATG13(designated as FIP200-4A mutant or FIP200 KI)does not produce comparable detrimental effects.This highlights the likely critical involvement of the non-canonical functions of FIP200,the mechanisms of which have remained elusive.Here,utilizing genetic mouse models,we demonstrated that FIP200 mediates non-canonical autophagic degradation of p62/sequestome1,primarily via TAX1BP1 in NSCs.Conditional deletion of Tax1bp1 in fip200hGFAP conditional knock-in(cKI)mice led to NSC deficiency,resembling the fip200hGFAP conditional knockout(cKO)mouse phenotype.Notably,reintroducing wild-type TAX1BP1 not only restored the maintenance of NSCs derived from tax1bp1-knockout fip200hGFAP cKI mice but also led to a marked reduction in p62 aggregate accumulation.Conversely,a TAX1BP1 mutant incapable of binding to FIP200 or NBR1/p62 failed to achieve this restoration.Furthermore,conditional deletion of Tax1bp1 in fip200hGFAP cKO mice exacerbated NSC deficiency and p62 aggregate accumulation compared to fip200hGFAP cKO mice.Collectively,these findings illustrate the essential role of the FIP200-TAX1BP1 axis in mediating the non-canonical autophagic degradation of p62 aggregates towards NSC maintenance and function,presenting novel therapeutic targets for neurodegenerative diseases.展开更多
基金Supported by The Deputyship for Research&Innovation,Ministry of Education in Saudi Arabia,for funding this research work through the project number ISP-24.,Jazan University,Jazan 82817,Saudi Arabia.
文摘BACKGROUND Obesity and type 2 diabetes mellitus(T2DM)are frequent co-occurring disorders that affect regular metabolic functions.Obesity has also been linked to an inc-reased risk of developing diabetes.Obesity and diabetes are on the rise,increa-sing healthcare costs and raising mortality rates.Research has revealed that the expression profile of microRNAs(miRNAs)changes as diabetes progresses.Fur-thermore,vitamin D may have an anti-obesity effect and inverse association with body weight and body mass index(BMI).Low vitamin D levels do not solely cause obesity,which could be a factor in the etiology of T2DM.METHODS This study included 210 participants,of which,82 were obese(BMI>30 kg/m2)without T2DM,28 were obese with T2DM,and 100 were healthy controls.BMI was evaluated and both fasting and postprandial blood glucose were used to confirm T2DM.Exosomal miRNA-200a and miRNA-200b expression were analyzed using real-time PCR using Taqman probes,and vitamin-D levels were evaluated using an electrochemiluminescence-based immunoassay technique.All data analyses were performed using SPSS 20.0 and GraphPad Prism 5 software.RESULTS Overall,a 2.20-and 4.40-fold increase in miRNA-200a and miRNA-200b expression was observed among participants compared to healthy controls.MiRNA-200a and miRNA-200b expression among obese participants increased 2.40-fold and 3.93-fold,respectively,while in obese T2DM participants these values were 2.67-fold,and 5.78-fold,respectively,and these differences were found to be statistically significant(P=0.02)(P<0.0001).Obese participants showed a vitamin D level of 34.27 ng/mL,while in obese-T2DM participants vitamin D level was 22.21 ng/mL(P<0.0001).Vitamin D was negatively correlated with miRNA-200a(r=-0.22,P=0.01)and miRNA-200b(r=-0.19,P=0.04).MiRNA-200a sensitivity was 75%,and specificity was 57%,with a cutoff value of 2.07-fold.MiRNA-200b sensitivity was 75%,and specificity was 71%with a cutoff value of 4.12-fold,suggesting that miRNA-200a and miRNA-200b with an increased expression of 2.07-and 4.12-fold could be predictive indicators for the risk of diabetes in obese participants.CONCLUSION MiRNA-200a and miRNA-200b were higher in diabetic obese participants vs non-diabetic obese participants,and insufficient vitamin D levels in obese T2DM participants may be involved in poor clinical outcome.
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.
基金National Natural Science Foundation of China(U2004138,81773132,81820108021)University Excellent Teaching Team of“Qinglan Project”in Jiangsu Province(2022-25)+1 种基金Henan Province Key Research and Development Project(232102521028)Excellent Youth Foundation of Henan Scientific Committee(21230040016)。
文摘Autophagy plays a pivotal role in diverse biological processes,including the maintenance and differentiation of neural stem cells(NSCs).Interestingly,while complete deletion of Fip200 severely impairs NSC maintenance and differentiation,inhibiting canonical autophagy via deletion of core genes,such as Atg5,Atg16l1,and Atg7,or blockade of canonical interactions between FIP200 and ATG13(designated as FIP200-4A mutant or FIP200 KI)does not produce comparable detrimental effects.This highlights the likely critical involvement of the non-canonical functions of FIP200,the mechanisms of which have remained elusive.Here,utilizing genetic mouse models,we demonstrated that FIP200 mediates non-canonical autophagic degradation of p62/sequestome1,primarily via TAX1BP1 in NSCs.Conditional deletion of Tax1bp1 in fip200hGFAP conditional knock-in(cKI)mice led to NSC deficiency,resembling the fip200hGFAP conditional knockout(cKO)mouse phenotype.Notably,reintroducing wild-type TAX1BP1 not only restored the maintenance of NSCs derived from tax1bp1-knockout fip200hGFAP cKI mice but also led to a marked reduction in p62 aggregate accumulation.Conversely,a TAX1BP1 mutant incapable of binding to FIP200 or NBR1/p62 failed to achieve this restoration.Furthermore,conditional deletion of Tax1bp1 in fip200hGFAP cKO mice exacerbated NSC deficiency and p62 aggregate accumulation compared to fip200hGFAP cKO mice.Collectively,these findings illustrate the essential role of the FIP200-TAX1BP1 axis in mediating the non-canonical autophagic degradation of p62 aggregates towards NSC maintenance and function,presenting novel therapeutic targets for neurodegenerative diseases.
