激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SIONFH)是由于糖皮质激素使用不当或过度而引起的髋关节疾病,发病机制尚未统一,临床疗效亦不佳。当前,没有效果明确的药物可以延缓疾病进程,而中医药治疗SIONFH在...激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SIONFH)是由于糖皮质激素使用不当或过度而引起的髋关节疾病,发病机制尚未统一,临床疗效亦不佳。当前,没有效果明确的药物可以延缓疾病进程,而中医药治疗SIONFH在临床上取得一定疗效。即便如此,仍未能完整的从分子生物及细胞生物学角度阐明中药治疗SIONFH的作用机制。转化生长因子-β(TGF-β)/骨形态发生蛋白(BMP)/Smad信号通路的转导是防治SIONFH的研究热点之一,故该文阐明了该信号通路的转导机制以及与SIONFH的联系,检索了基于该通路治疗SIONFH的全部中药及复方并阐述其影响机制。基于中医对SIONFH的认识,现临床上使用补肝肾强筋骨以及活血祛瘀通络类的方药治疗SIONFH,且具有良好的疗效。中药通过调控该通路,可刺激骨髓间充质干细胞成骨分化,降低破骨细胞含量,减少脂肪生成,改善微循环,抗氧化损伤,促进股骨头内血管新生,从而促进股骨头损伤的修复。现基于TGF-β/BMP/Smad信号通路对中医药治疗SIONFH的研究进展做一综述,期许为中医药治疗SIONFH提供理论依据及参考。展开更多
旨在探究不同FecB基因型对绵羊卵泡中BMP/SMAD通路活性和蛋白表达的影响;揭示成熟大卵泡和小卵泡之间BMP/SMAD通路活性和蛋白表达的差异。本研究采用TaqMan分型方法筛选出不同FecB基因型的母羊,同期发情后取卵泡期成熟卵泡和黄体期卵巢...旨在探究不同FecB基因型对绵羊卵泡中BMP/SMAD通路活性和蛋白表达的影响;揭示成熟大卵泡和小卵泡之间BMP/SMAD通路活性和蛋白表达的差异。本研究采用TaqMan分型方法筛选出不同FecB基因型的母羊,同期发情后取卵泡期成熟卵泡和黄体期卵巢表面小卵泡,利用免疫印迹法(Western blot)测定BMP/SMAD通路相关蛋白表达水平和通路活性。结果表明,对于小卵泡组,FecB突变型卵泡中骨形态发生蛋白1B型受体(bone morphogenetic protein receptor type 1B,BMPR1B)表达量显著高于野生型卵泡(P<0.05),但SMAD家族成员4(SMAD family member 4,SMAD4)表达量和SMAD1/5/9的磷酸化水平显著低于野生型卵泡(P<0.05);对于成熟大卵泡组,FecB突变型卵泡中FKBP脯氨酰异构酶1A(FKBP prolyl isomerase 1A,FKBP1A)和SMAD4表达量显著低于野生型卵泡(P<0.05),Ⅰ型受体(BMPR1B)和Ⅱ型受体(BMPR2)的蛋白表达量及SMAD1/5/9的磷酸化水平在两种基因型之间未显示出显著差异。另一方面,对比FecB突变型小卵泡和成熟大卵泡发现:成熟大卵泡中BMPR1B和SMAD4蛋白表达量和SMAD1/5/9磷酸化程度显著高于小卵泡(P<0.05)。上述结果表明,由于SMAD4表达量的下降,FecB突变型大、小卵泡中结合到基因组靶区域的SMAD4-SMAD1/5/9蛋白复合物均相对较少,将导致通路活性降低,而且由于小卵泡中较低的SMAD1/5/9磷酸化水平,其通路活性更低。另外,绵羊突变型卵泡生长发育成熟后BMP/SMAD通路活性显著增强。展开更多
目的:研究丹皮酚对强直性脊柱炎(AS)模型小鼠分泌型糖蛋白(Wnt)和骨形态发生蛋白(BMP)/细胞信号转导分子(Smad)通路的影响,探讨丹皮酚防治AS的机制。方法:将40只小鼠随机分为正常组、模型组、柳氮磺吡啶组(阳性对照,9 mg/kg)和丹皮酚组(...目的:研究丹皮酚对强直性脊柱炎(AS)模型小鼠分泌型糖蛋白(Wnt)和骨形态发生蛋白(BMP)/细胞信号转导分子(Smad)通路的影响,探讨丹皮酚防治AS的机制。方法:将40只小鼠随机分为正常组、模型组、柳氮磺吡啶组(阳性对照,9 mg/kg)和丹皮酚组(3 mg/kg),每组10只。除正常组外的其余各组小鼠均采用完全弗氏佐剂+蛋白聚糖腹腔注射法复制AS模型。各给药组小鼠在成模后灌胃相应药物,正常组和模型组小鼠灌胃等体积蒸馏水,每天给药1次,连续20 d。末次给药后处死小鼠,透射电镜下观察各组小鼠骶髂关节滑膜细胞超微病理结构变化,采用酶联免疫吸附法检测小鼠血清中肿瘤坏死因子α(TNF-α)、Wnt信号通路病理性骨化相关因子(DKK-1)含量,实时荧光定量聚合酶链式反应法检测小鼠滑膜组织中骨形态发生蛋白2(BMP-2)、细胞内核心结合因子α1(Cbfα1)、Smad1 m RNA表达。结果:与正常组比较,模型组小鼠血清中TNF-α含量明显增加、DKK-1含量明显减少,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显升高,差异均有统计学意义(P<0.05或P<0.01);电镜下可见模型组小鼠滑膜细胞增生,排列紊乱,分泌活性细胞器分泌亢进,细胞间隙增宽。与模型组比较,柳氮磺吡啶组和丹皮酚组小鼠血清中TNF-α含量均明显减少,丹皮酚组小鼠血清中DKK-1含量明显增加,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显降低,差异均有统计学意义(P<0.05或P<0.01);电镜下可见丹皮酚组小鼠滑膜细胞线粒体、溶酶体、粗面内质网的形态明显改善。结论:丹皮酚防治AS的机制可能与降低血清中TNF-α含量、升高血清中DKK-1含量,下调滑膜细胞中BMP-2、Cbfα1和Smad1 m RNA表达,抑制Wnt和BMP/Smad骨化相关信号转导通路逆转滑膜细胞成骨分化有关。展开更多
Background:Osteoporosis is a chronic bone disease characterized by bone loss and decreased bone strength.However,current anti-resorptive drugs carry a risk of various complications.The deep learning-based efficacy pre...Background:Osteoporosis is a chronic bone disease characterized by bone loss and decreased bone strength.However,current anti-resorptive drugs carry a risk of various complications.The deep learning-based efficacy prediction system(DLEPS)is a forecasting tool that can effectively compete in drug screening and prediction based on gene expression changes.This study aimed to explore the protective effect and potential mechanisms of cinobufotalin(CB),a traditional Chinese medicine(TCM),on bone loss.Methods:DLEPS was employed for screening anti-osteoporotic agents according to gene profile changes in primary osteoporosis.Micro-CT,histological and morphological analysis were applied for the bone protective detection of CB,and the osteogenic differentiation/function in human bone marrow mesenchymal stem cells(hBMMSCs)were also investigated.The underlying mechanism was verified using qRT-PCR,Western blot(WB),immunofluorescence(IF),etc.Results:A safe concentration(0.25mg/kg in vivo,0.05μM in vitro)of CB could effectively preserve bone mass in estrogen deficiency-induced bone loss and promote osteogenic differentiation/function of hBMMSCs.Both BMPs/SMAD and Wnt/β-catenin signaling pathways participated in CB-induced osteogenic differentiation,further regulating the expression of osteogenesis-associated factors,and ultimately promoting osteogenesis.Conclusion:Our study demonstrated that CB could significantly reverse estrogen deficiency-induced bone loss,further promoting osteogenic differentiation/function of hBMMSCs,with BMPs/SMAD and Wnt/β-catenin signaling pathways involved.展开更多
文摘激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SIONFH)是由于糖皮质激素使用不当或过度而引起的髋关节疾病,发病机制尚未统一,临床疗效亦不佳。当前,没有效果明确的药物可以延缓疾病进程,而中医药治疗SIONFH在临床上取得一定疗效。即便如此,仍未能完整的从分子生物及细胞生物学角度阐明中药治疗SIONFH的作用机制。转化生长因子-β(TGF-β)/骨形态发生蛋白(BMP)/Smad信号通路的转导是防治SIONFH的研究热点之一,故该文阐明了该信号通路的转导机制以及与SIONFH的联系,检索了基于该通路治疗SIONFH的全部中药及复方并阐述其影响机制。基于中医对SIONFH的认识,现临床上使用补肝肾强筋骨以及活血祛瘀通络类的方药治疗SIONFH,且具有良好的疗效。中药通过调控该通路,可刺激骨髓间充质干细胞成骨分化,降低破骨细胞含量,减少脂肪生成,改善微循环,抗氧化损伤,促进股骨头内血管新生,从而促进股骨头损伤的修复。现基于TGF-β/BMP/Smad信号通路对中医药治疗SIONFH的研究进展做一综述,期许为中医药治疗SIONFH提供理论依据及参考。
文摘旨在探究不同FecB基因型对绵羊卵泡中BMP/SMAD通路活性和蛋白表达的影响;揭示成熟大卵泡和小卵泡之间BMP/SMAD通路活性和蛋白表达的差异。本研究采用TaqMan分型方法筛选出不同FecB基因型的母羊,同期发情后取卵泡期成熟卵泡和黄体期卵巢表面小卵泡,利用免疫印迹法(Western blot)测定BMP/SMAD通路相关蛋白表达水平和通路活性。结果表明,对于小卵泡组,FecB突变型卵泡中骨形态发生蛋白1B型受体(bone morphogenetic protein receptor type 1B,BMPR1B)表达量显著高于野生型卵泡(P<0.05),但SMAD家族成员4(SMAD family member 4,SMAD4)表达量和SMAD1/5/9的磷酸化水平显著低于野生型卵泡(P<0.05);对于成熟大卵泡组,FecB突变型卵泡中FKBP脯氨酰异构酶1A(FKBP prolyl isomerase 1A,FKBP1A)和SMAD4表达量显著低于野生型卵泡(P<0.05),Ⅰ型受体(BMPR1B)和Ⅱ型受体(BMPR2)的蛋白表达量及SMAD1/5/9的磷酸化水平在两种基因型之间未显示出显著差异。另一方面,对比FecB突变型小卵泡和成熟大卵泡发现:成熟大卵泡中BMPR1B和SMAD4蛋白表达量和SMAD1/5/9磷酸化程度显著高于小卵泡(P<0.05)。上述结果表明,由于SMAD4表达量的下降,FecB突变型大、小卵泡中结合到基因组靶区域的SMAD4-SMAD1/5/9蛋白复合物均相对较少,将导致通路活性降低,而且由于小卵泡中较低的SMAD1/5/9磷酸化水平,其通路活性更低。另外,绵羊突变型卵泡生长发育成熟后BMP/SMAD通路活性显著增强。
文摘目的:研究丹皮酚对强直性脊柱炎(AS)模型小鼠分泌型糖蛋白(Wnt)和骨形态发生蛋白(BMP)/细胞信号转导分子(Smad)通路的影响,探讨丹皮酚防治AS的机制。方法:将40只小鼠随机分为正常组、模型组、柳氮磺吡啶组(阳性对照,9 mg/kg)和丹皮酚组(3 mg/kg),每组10只。除正常组外的其余各组小鼠均采用完全弗氏佐剂+蛋白聚糖腹腔注射法复制AS模型。各给药组小鼠在成模后灌胃相应药物,正常组和模型组小鼠灌胃等体积蒸馏水,每天给药1次,连续20 d。末次给药后处死小鼠,透射电镜下观察各组小鼠骶髂关节滑膜细胞超微病理结构变化,采用酶联免疫吸附法检测小鼠血清中肿瘤坏死因子α(TNF-α)、Wnt信号通路病理性骨化相关因子(DKK-1)含量,实时荧光定量聚合酶链式反应法检测小鼠滑膜组织中骨形态发生蛋白2(BMP-2)、细胞内核心结合因子α1(Cbfα1)、Smad1 m RNA表达。结果:与正常组比较,模型组小鼠血清中TNF-α含量明显增加、DKK-1含量明显减少,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显升高,差异均有统计学意义(P<0.05或P<0.01);电镜下可见模型组小鼠滑膜细胞增生,排列紊乱,分泌活性细胞器分泌亢进,细胞间隙增宽。与模型组比较,柳氮磺吡啶组和丹皮酚组小鼠血清中TNF-α含量均明显减少,丹皮酚组小鼠血清中DKK-1含量明显增加,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显降低,差异均有统计学意义(P<0.05或P<0.01);电镜下可见丹皮酚组小鼠滑膜细胞线粒体、溶酶体、粗面内质网的形态明显改善。结论:丹皮酚防治AS的机制可能与降低血清中TNF-α含量、升高血清中DKK-1含量,下调滑膜细胞中BMP-2、Cbfα1和Smad1 m RNA表达,抑制Wnt和BMP/Smad骨化相关信号转导通路逆转滑膜细胞成骨分化有关。
基金Beijing Natural Science Foundation,Grant/Award Number:L222145 and L222030Emerging Engineering Interdisciplinary Project and the Fundamental Research Funds for the Central Universities,Grant/Award Number:PKU2022XGK008Peking University Medicine Fund of Fostering Young Scholars’Scientific&Technological Innovation,Grant/Award Number:BMU2022PY010。
文摘Background:Osteoporosis is a chronic bone disease characterized by bone loss and decreased bone strength.However,current anti-resorptive drugs carry a risk of various complications.The deep learning-based efficacy prediction system(DLEPS)is a forecasting tool that can effectively compete in drug screening and prediction based on gene expression changes.This study aimed to explore the protective effect and potential mechanisms of cinobufotalin(CB),a traditional Chinese medicine(TCM),on bone loss.Methods:DLEPS was employed for screening anti-osteoporotic agents according to gene profile changes in primary osteoporosis.Micro-CT,histological and morphological analysis were applied for the bone protective detection of CB,and the osteogenic differentiation/function in human bone marrow mesenchymal stem cells(hBMMSCs)were also investigated.The underlying mechanism was verified using qRT-PCR,Western blot(WB),immunofluorescence(IF),etc.Results:A safe concentration(0.25mg/kg in vivo,0.05μM in vitro)of CB could effectively preserve bone mass in estrogen deficiency-induced bone loss and promote osteogenic differentiation/function of hBMMSCs.Both BMPs/SMAD and Wnt/β-catenin signaling pathways participated in CB-induced osteogenic differentiation,further regulating the expression of osteogenesis-associated factors,and ultimately promoting osteogenesis.Conclusion:Our study demonstrated that CB could significantly reverse estrogen deficiency-induced bone loss,further promoting osteogenic differentiation/function of hBMMSCs,with BMPs/SMAD and Wnt/β-catenin signaling pathways involved.
