Objective:To investigate the antifibrotic effects of curcumin in a transverse aortic constriction(TAC)mouse model and elucidate its molecular mechanisms.Methods:Male C57BL/6 mice underwent TAC and received vehicle,low...Objective:To investigate the antifibrotic effects of curcumin in a transverse aortic constriction(TAC)mouse model and elucidate its molecular mechanisms.Methods:Male C57BL/6 mice underwent TAC and received vehicle,low-dose curcumin(50 mg/kg),high-dose curcumin(200 mg/kg),high-dose curcumin plus a scrambled control antagomir,or high-dose curcumin plus anti-miR-29b treatments.Cardiac function was assessed by echocardiography.Fibrosis was evaluated by histology,collagen volume fraction,and hydroxyproline content.Expression of miR-29b,HDAC4,and fibrosis-related markers(Col1a1,Col3a1,TGF-β1)was measured by quantitative RT-PCR and Western blotting assays.Myocardial procollagen type I carboxy-terminal propeptide was determined by ELISA,and HDAC4-specific enzymatic activity was assayed using a fluorogenic kit.Results:Curcumin improved cardiac function,reduced fibrosis,restored miR-29b expression,and suppressed HDAC4 expression and activity in a dose-dependent manner.Furthermore,curcumin decreased myocardial procollagen type I carboxy-terminal propeptide levels,confirming reduced collagen synthesis.Anti-miR-29b administration partially abrogated the antifibrotic and cardioprotective effects of curcumin.Conclusions:Curcumin attenuates pressure overload-induced cardiac fibrosis and dysfunction in a TAC mouse model via modulation of the miR-29b/HDAC4 axis and suppression of collagen synthesis.展开更多
目的研究miR-145-3p能否调控人牙周膜成纤维细胞(HPDLFs)的自噬及其可能存在的作用机制。方法收集12~18岁因正畸需要减数而拔除的健康前磨牙,采集和培养HPDLFs,并进行鉴定。将miR-145-3p类似物(miR-145-3p-mimics组)、miR-145-3p抑制物(...目的研究miR-145-3p能否调控人牙周膜成纤维细胞(HPDLFs)的自噬及其可能存在的作用机制。方法收集12~18岁因正畸需要减数而拔除的健康前磨牙,采集和培养HPDLFs,并进行鉴定。将miR-145-3p类似物(miR-145-3p-mimics组)、miR-145-3p抑制物(miR-145-3p-inhibitor组)、阴性对照(miR-145-3p normal control,miR-145-3p-NC组)以及带有GFP-LC3的质粒转染至HPDLFs,24 h后荧光显微镜观察荧光发生情况。双荧光素酶报告实验验证miR-145-3p与HDAC4的靶向关系,Western Blot检测各转染组细胞内HDAC4、Beclin-1、P62和LC3的蛋白表达。结果免疫组化结果表明,波形蛋白为阳性染色而角蛋白呈阴性染色,证明其为无混杂细胞的HPDLFs。转染结果显示,miR-145-3p-mimics组的细胞内荧光强度最高,有大量自噬体形成,而miR-145-3p-inhibitor组荧光最弱。双荧光素酶报告实验证实miR-145-3p靶向抑制HPDLFs细胞中HDAC4的表达。4组细胞中miR-145-3p-inhibitor组P62蛋白表达最高(P<0.05),而其他3组差异无统计学意义(P>0.05);miR-145-3p-mimics组细胞内Beclin-1和LC3蛋白表达最高(P<0.05),其他3组差异无统计学意义(P>0.05)。结论miR-145-3p在HPDLFs中可调控自噬,这种作用可能是通过靶向抑制HDAC4表达实现的。展开更多
This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 p...This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 pg/kg, i.v. after ending occlusion） significantly reduced neurological deficit scores, infarct volumes, and brain water contents after 1.5 h MCAO and 24 h reperfusion. 002C-3 （75 150μg/kg） decreased the exudation of Evans blue from brain capillaries. 002C-3 （100 μg/kg） significantly inhibited the activity of MMP-9 and MMP-2 in the injured hemisphere. 002C-3 decreased the expression of autophagy-associated proteins, Beclin-1 and LC3B-Ⅱ, and increased the level of p62 in injured hemisphere. 002C-3 （100 pg/kg） significantly increased the expression of p-CaMKIV and p-HDAC4 in injured hemisphere. In conclusion, 002C-3 shows a neuroprotective effect on tMCAO injury in mice, and its mechanisms may be associated with alleviation of blood-brain barrier damage caused by the activation of MMPs, inhibition of autophagy, and stimulation of calcium signals related to cell survival. These findings suggest that 002C-3 is a neuroprotective agent that acts on multiple pathways.展开更多
基金supported by China International Medical Foundation(Z-2019-42-1908-4)Natural Science Basic Research Program of Shaanxi Province(2019JM-440).
文摘Objective:To investigate the antifibrotic effects of curcumin in a transverse aortic constriction(TAC)mouse model and elucidate its molecular mechanisms.Methods:Male C57BL/6 mice underwent TAC and received vehicle,low-dose curcumin(50 mg/kg),high-dose curcumin(200 mg/kg),high-dose curcumin plus a scrambled control antagomir,or high-dose curcumin plus anti-miR-29b treatments.Cardiac function was assessed by echocardiography.Fibrosis was evaluated by histology,collagen volume fraction,and hydroxyproline content.Expression of miR-29b,HDAC4,and fibrosis-related markers(Col1a1,Col3a1,TGF-β1)was measured by quantitative RT-PCR and Western blotting assays.Myocardial procollagen type I carboxy-terminal propeptide was determined by ELISA,and HDAC4-specific enzymatic activity was assayed using a fluorogenic kit.Results:Curcumin improved cardiac function,reduced fibrosis,restored miR-29b expression,and suppressed HDAC4 expression and activity in a dose-dependent manner.Furthermore,curcumin decreased myocardial procollagen type I carboxy-terminal propeptide levels,confirming reduced collagen synthesis.Anti-miR-29b administration partially abrogated the antifibrotic and cardioprotective effects of curcumin.Conclusions:Curcumin attenuates pressure overload-induced cardiac fibrosis and dysfunction in a TAC mouse model via modulation of the miR-29b/HDAC4 axis and suppression of collagen synthesis.
文摘目的研究miR-145-3p能否调控人牙周膜成纤维细胞(HPDLFs)的自噬及其可能存在的作用机制。方法收集12~18岁因正畸需要减数而拔除的健康前磨牙,采集和培养HPDLFs,并进行鉴定。将miR-145-3p类似物(miR-145-3p-mimics组)、miR-145-3p抑制物(miR-145-3p-inhibitor组)、阴性对照(miR-145-3p normal control,miR-145-3p-NC组)以及带有GFP-LC3的质粒转染至HPDLFs,24 h后荧光显微镜观察荧光发生情况。双荧光素酶报告实验验证miR-145-3p与HDAC4的靶向关系,Western Blot检测各转染组细胞内HDAC4、Beclin-1、P62和LC3的蛋白表达。结果免疫组化结果表明,波形蛋白为阳性染色而角蛋白呈阴性染色,证明其为无混杂细胞的HPDLFs。转染结果显示,miR-145-3p-mimics组的细胞内荧光强度最高,有大量自噬体形成,而miR-145-3p-inhibitor组荧光最弱。双荧光素酶报告实验证实miR-145-3p靶向抑制HPDLFs细胞中HDAC4的表达。4组细胞中miR-145-3p-inhibitor组P62蛋白表达最高(P<0.05),而其他3组差异无统计学意义(P>0.05);miR-145-3p-mimics组细胞内Beclin-1和LC3蛋白表达最高(P<0.05),其他3组差异无统计学意义(P>0.05)。结论miR-145-3p在HPDLFs中可调控自噬,这种作用可能是通过靶向抑制HDAC4表达实现的。
基金National Natural Science Foundation of China(Grant No.81302763,81573333)Beijing Natural Science Foundation(Grant No.7144218)
文摘This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 pg/kg, i.v. after ending occlusion） significantly reduced neurological deficit scores, infarct volumes, and brain water contents after 1.5 h MCAO and 24 h reperfusion. 002C-3 （75 150μg/kg） decreased the exudation of Evans blue from brain capillaries. 002C-3 （100 μg/kg） significantly inhibited the activity of MMP-9 and MMP-2 in the injured hemisphere. 002C-3 decreased the expression of autophagy-associated proteins, Beclin-1 and LC3B-Ⅱ, and increased the level of p62 in injured hemisphere. 002C-3 （100 pg/kg） significantly increased the expression of p-CaMKIV and p-HDAC4 in injured hemisphere. In conclusion, 002C-3 shows a neuroprotective effect on tMCAO injury in mice, and its mechanisms may be associated with alleviation of blood-brain barrier damage caused by the activation of MMPs, inhibition of autophagy, and stimulation of calcium signals related to cell survival. These findings suggest that 002C-3 is a neuroprotective agent that acts on multiple pathways.
