目的基于骨组织Dlx5启动子甲基化水平,探索补肾中药复方对大鼠去卵巢骨质疏松症的疗效机制。方法去卵巢建立绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠模型,分为正常组、模型组、补肾中药复方组、仙灵骨葆阳性对照组。灌...目的基于骨组织Dlx5启动子甲基化水平,探索补肾中药复方对大鼠去卵巢骨质疏松症的疗效机制。方法去卵巢建立绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠模型,分为正常组、模型组、补肾中药复方组、仙灵骨葆阳性对照组。灌胃14周后,X线吸收测量法检测骨密度,质谱法检测Dlx5启动子甲基化水平。结果与正常组比较,模型组第1~6腰椎骨密度明显降低(P<0.01),骨组织Dlx5启动子-470 bp^-4 bp CpG2.3甲基化水平明显升高(P<0.01)。②与模型组比较,补肾中药复方组、仙灵骨葆阳性对照组第1~6腰椎骨密度明显升高(P<0.05),骨组织Dlx5启动子-470 bp^-4 bp CpG1甲基化水平明显降低(P<0.05)。结论补肾中药复方通过降低骨组织Dlx5启动子甲基化水平的表观遗传学机制,有效防治PMOP。展开更多
Osteoarthritis(OA)has been considered non-reversible as articular cartilage wears down with limited repair capacity.Enhanced chondrocyte hypertrophy and increased type X collagen gene(COL10A1)expression have been asso...Osteoarthritis(OA)has been considered non-reversible as articular cartilage wears down with limited repair capacity.Enhanced chondrocyte hypertrophy and increased type X collagen gene(COL10A1)expression have been associated with OA.Therefore,regulators controlling collagen X expression and chondrocyte hypertrophy may play a role in OA intervention.Here,we investigated how Distal-less homeobox 5(DLX5),the distal-less homeobox family member,controls murine Col10a1 gene expression and chondrocyte hypertrophy in chondrogenic cell models and its role in a murine OA model.Through qRT-PCR and Western blot analyses,we detected significantly increased levels of COL10A1 and DLX5 in hypertrophic MCT and ATDC5 cells compared to their proliferative stage.Forced expression of Dlx5 further increases,while knockdown of Dlx5 decreases COL10A1 expression in hypertrophic MCT cells.We have performed dual-luciferase reporter and ChiP assays and demonstrated that DLX5 promotes reporter activity through direct interaction with Col10a1 cis-enhancer.We established a murine OA model and detected markedly increased COL10A1 and DLX5 in the articular cartilage and subchondral bone of the OA mice compared with the controls.Notably,forced overexpression of DLX5 in hypertrophic MCT cells up-regulates RUNX2,and adjacent DLX5 and RUNX2 binding sites have previously been found within the Col10a1 cis-enhancer.Together,our data suggest that DLX5 may cooperate with RUNX2 to control cell-specific Col10a1 expression and chondrocyte hypertrophy and is involved in OA pathogenesis.展开更多
文摘目的基于骨组织Dlx5启动子甲基化水平,探索补肾中药复方对大鼠去卵巢骨质疏松症的疗效机制。方法去卵巢建立绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠模型,分为正常组、模型组、补肾中药复方组、仙灵骨葆阳性对照组。灌胃14周后,X线吸收测量法检测骨密度,质谱法检测Dlx5启动子甲基化水平。结果与正常组比较,模型组第1~6腰椎骨密度明显降低(P<0.01),骨组织Dlx5启动子-470 bp^-4 bp CpG2.3甲基化水平明显升高(P<0.01)。②与模型组比较,补肾中药复方组、仙灵骨葆阳性对照组第1~6腰椎骨密度明显升高(P<0.05),骨组织Dlx5启动子-470 bp^-4 bp CpG1甲基化水平明显降低(P<0.05)。结论补肾中药复方通过降低骨组织Dlx5启动子甲基化水平的表观遗传学机制,有效防治PMOP。
基金supported by the Jiangsu Provincial Key Research and Development Program(China)(No.BE2020679 to Q.Z.)the Innovation Team(leader)of Jiangsu Province,China(2017,QZ)the National Natural Science Foundation of China(No.82001576 to L.Q.)。
文摘Osteoarthritis(OA)has been considered non-reversible as articular cartilage wears down with limited repair capacity.Enhanced chondrocyte hypertrophy and increased type X collagen gene(COL10A1)expression have been associated with OA.Therefore,regulators controlling collagen X expression and chondrocyte hypertrophy may play a role in OA intervention.Here,we investigated how Distal-less homeobox 5(DLX5),the distal-less homeobox family member,controls murine Col10a1 gene expression and chondrocyte hypertrophy in chondrogenic cell models and its role in a murine OA model.Through qRT-PCR and Western blot analyses,we detected significantly increased levels of COL10A1 and DLX5 in hypertrophic MCT and ATDC5 cells compared to their proliferative stage.Forced expression of Dlx5 further increases,while knockdown of Dlx5 decreases COL10A1 expression in hypertrophic MCT cells.We have performed dual-luciferase reporter and ChiP assays and demonstrated that DLX5 promotes reporter activity through direct interaction with Col10a1 cis-enhancer.We established a murine OA model and detected markedly increased COL10A1 and DLX5 in the articular cartilage and subchondral bone of the OA mice compared with the controls.Notably,forced overexpression of DLX5 in hypertrophic MCT cells up-regulates RUNX2,and adjacent DLX5 and RUNX2 binding sites have previously been found within the Col10a1 cis-enhancer.Together,our data suggest that DLX5 may cooperate with RUNX2 to control cell-specific Col10a1 expression and chondrocyte hypertrophy and is involved in OA pathogenesis.
