Objectives The discovery of novel molecular targets to enhance the osteogenesis of human bone marrow-derived mesenchymal stem cells(H-BMSCs)represents a promising strategy for preventing and treating osteoporosis.Thus...Objectives The discovery of novel molecular targets to enhance the osteogenesis of human bone marrow-derived mesenchymal stem cells(H-BMSCs)represents a promising strategy for preventing and treating osteoporosis.Thus,the primary objective of this study is to elucidate the mechanisms by which long non-coding RNA FOXD2-AS1(lncRNA FOXD2-AS1)regulates early osteogenic differentiation in H-BMSCs,thereby identifying potential therapeutic targets.Methods Lentivirus-mediated vectors were constructed to either overexpress or silence FOXD2-AS1 in H-BMSCs.The effects of FOXD2-AS1 on osteogenesis were subsequently assessed by analyzing osteogenic marker expression and alkaline phosphatase(ALP)staining.To clarify the role of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway in this process,AG490 inhibitor(a JAK2/STAT3 pathway inhibitor)and knockdown of STAT3 were used to investigate the mechanisms of FOXD2-AS1.Results FOXD2-AS1 overexpression increased ALP activity and osteogenic marker expression,while its knockdown had the opposite effects.From a mechanistic perspective,FOXD2-AS1 overexpression promoted JAK2 and STAT3 phosphorylation,whereas its suppression attenuated their activation.Also,the osteogenic increase induced by FOXD2-AS1 overexpression was reversed by AG490 treatment or STAT3 silencing,indicating that the pathway plays a role in this process.Conclusion FOXD2-AS1 was identified as a novel genetic switch driving osteogenic commitment via JAK2/STAT3 activation,revealing a new regulatory mechanism and a potential therapeutic target for osteoporosis.展开更多
目的探讨银杏内酯B通过激活免疫性血小板减少症(ITP)哺乳动物雷帕霉素靶蛋白(mTOR)信号通路调节长链非编码RNA人浆细胞瘤转化迁移基因1(lncRNA PVT1)促进调节性T淋巴细胞/辅助性T淋巴细胞17(Treg/Th17)免疫平衡而抑制炎症反应的机制。...目的探讨银杏内酯B通过激活免疫性血小板减少症(ITP)哺乳动物雷帕霉素靶蛋白(mTOR)信号通路调节长链非编码RNA人浆细胞瘤转化迁移基因1(lncRNA PVT1)促进调节性T淋巴细胞/辅助性T淋巴细胞17(Treg/Th17)免疫平衡而抑制炎症反应的机制。方法选用80只雄性SD小鼠,采用腹腔注射抗小鼠血小板血清方法建立模型,随机分为正常组、模型组、低剂量组、高剂量组,每组20只,连续给药14天。实时荧光定量PCR(qRT-PCR)检测血清lncRNA PVT1表达,流式细胞仪检测Treg/Th17细胞,Western Blot检测磷脂酰肌醇-3-激酶(PI3K)、Akt、mTOR蛋白表达。检测血清白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)和全血血小板计数(PLT)。结果与正常组比较,模型组小鼠血清lncRNA PVT1、Th17、IFN-γ升高,Treg、Treg/Th17、PI3K、Akt、mTOR蛋白、IL-4、PLT降低,差异具有统计学意义(t=2.510~54.899,均P<0.05);与模型组比较,低剂量组、高剂量组lncRNA PVT1(1.99±0.14、1.25±0.11 vs 2.39±0.15)、Th17(1.76%±0.32%、0.87%±0.04%vs 5.28%±1.21%)、IFN-γ(7.65±0.28pg/ml、6.84±0.33pg/ml vs 8.45±0.36 pg/ml)均降低(t_(低剂量组)=8.718、8.782、3.292,t_(高剂量组)=27.408、9.789、6.542),Treg(3.46%±0.43%、4.77%±0.51%vs 2.41%±0.44%)、Treg/Th17(1.98±0.16、4.24±1.02 vs 0.45±0.05)、PI3K(0.88±0.08、1.22±0.21 vs 0.45±0.05)、Akt(0.66±0.07、1.11±0.11 vs 0.21±0.02)、mTOR蛋白(0.70±0.08、1.21±0.13 vs 0.45±0.06)、IL-4(12.28±1.28pg/ml、13.08±1.01pg/ml vs 11.45±1.05pg/ml)、PLT[(526.99±50.34)×10^(9)/L、(880.37±52.78)×10^(9)/L vs(218.58±50.35)×10^(9)/L]均升高(t_(低剂量组)=4.943~27.643,t_(高剂量组)=7.766~40.818),差异具有统计学意义(均P<0.05)。与低剂量组比较,高剂量组lncRNA PVT1、Th17、IFN-γ降低,Treg、Treg/Th17、PI3K、Akt、mTOR蛋白、IL-4、PLT升高,差异具有统计学意义(t=2.411~21.667,均P<0.05)。结论银杏内酯B可通过激活mTOR信号通路,调节lncRNA PVT1促进Treg/Th17免疫平衡,抑制ITP小鼠炎症反应,提升血小板数量。展开更多
基金supported by the Natural Science Foundation of Hubei Province of China(Grant No.2023AFB671)the National Natural Science Foundation of China(Grant Nos.82360177 and 82560182)+1 种基金the Key Project of Jiangxi Provincial Natural Science Foundation(Grant No.20224ACB206011)“Xuncheng Talents”Project in Jiujiang City,Jiangxi Province(Grant No.JJXC2023071).
文摘Objectives The discovery of novel molecular targets to enhance the osteogenesis of human bone marrow-derived mesenchymal stem cells(H-BMSCs)represents a promising strategy for preventing and treating osteoporosis.Thus,the primary objective of this study is to elucidate the mechanisms by which long non-coding RNA FOXD2-AS1(lncRNA FOXD2-AS1)regulates early osteogenic differentiation in H-BMSCs,thereby identifying potential therapeutic targets.Methods Lentivirus-mediated vectors were constructed to either overexpress or silence FOXD2-AS1 in H-BMSCs.The effects of FOXD2-AS1 on osteogenesis were subsequently assessed by analyzing osteogenic marker expression and alkaline phosphatase(ALP)staining.To clarify the role of the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway in this process,AG490 inhibitor(a JAK2/STAT3 pathway inhibitor)and knockdown of STAT3 were used to investigate the mechanisms of FOXD2-AS1.Results FOXD2-AS1 overexpression increased ALP activity and osteogenic marker expression,while its knockdown had the opposite effects.From a mechanistic perspective,FOXD2-AS1 overexpression promoted JAK2 and STAT3 phosphorylation,whereas its suppression attenuated their activation.Also,the osteogenic increase induced by FOXD2-AS1 overexpression was reversed by AG490 treatment or STAT3 silencing,indicating that the pathway plays a role in this process.Conclusion FOXD2-AS1 was identified as a novel genetic switch driving osteogenic commitment via JAK2/STAT3 activation,revealing a new regulatory mechanism and a potential therapeutic target for osteoporosis.
文摘目的探讨银杏内酯B通过激活免疫性血小板减少症(ITP)哺乳动物雷帕霉素靶蛋白(mTOR)信号通路调节长链非编码RNA人浆细胞瘤转化迁移基因1(lncRNA PVT1)促进调节性T淋巴细胞/辅助性T淋巴细胞17(Treg/Th17)免疫平衡而抑制炎症反应的机制。方法选用80只雄性SD小鼠,采用腹腔注射抗小鼠血小板血清方法建立模型,随机分为正常组、模型组、低剂量组、高剂量组,每组20只,连续给药14天。实时荧光定量PCR(qRT-PCR)检测血清lncRNA PVT1表达,流式细胞仪检测Treg/Th17细胞,Western Blot检测磷脂酰肌醇-3-激酶(PI3K)、Akt、mTOR蛋白表达。检测血清白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)和全血血小板计数(PLT)。结果与正常组比较,模型组小鼠血清lncRNA PVT1、Th17、IFN-γ升高,Treg、Treg/Th17、PI3K、Akt、mTOR蛋白、IL-4、PLT降低,差异具有统计学意义(t=2.510~54.899,均P<0.05);与模型组比较,低剂量组、高剂量组lncRNA PVT1(1.99±0.14、1.25±0.11 vs 2.39±0.15)、Th17(1.76%±0.32%、0.87%±0.04%vs 5.28%±1.21%)、IFN-γ(7.65±0.28pg/ml、6.84±0.33pg/ml vs 8.45±0.36 pg/ml)均降低(t_(低剂量组)=8.718、8.782、3.292,t_(高剂量组)=27.408、9.789、6.542),Treg(3.46%±0.43%、4.77%±0.51%vs 2.41%±0.44%)、Treg/Th17(1.98±0.16、4.24±1.02 vs 0.45±0.05)、PI3K(0.88±0.08、1.22±0.21 vs 0.45±0.05)、Akt(0.66±0.07、1.11±0.11 vs 0.21±0.02)、mTOR蛋白(0.70±0.08、1.21±0.13 vs 0.45±0.06)、IL-4(12.28±1.28pg/ml、13.08±1.01pg/ml vs 11.45±1.05pg/ml)、PLT[(526.99±50.34)×10^(9)/L、(880.37±52.78)×10^(9)/L vs(218.58±50.35)×10^(9)/L]均升高(t_(低剂量组)=4.943~27.643,t_(高剂量组)=7.766~40.818),差异具有统计学意义(均P<0.05)。与低剂量组比较,高剂量组lncRNA PVT1、Th17、IFN-γ降低,Treg、Treg/Th17、PI3K、Akt、mTOR蛋白、IL-4、PLT升高,差异具有统计学意义(t=2.411~21.667,均P<0.05)。结论银杏内酯B可通过激活mTOR信号通路,调节lncRNA PVT1促进Treg/Th17免疫平衡,抑制ITP小鼠炎症反应,提升血小板数量。
