Objective Cardiac fibroblasts(CFs)proliferation and extracellular matrix deposition are important features of cardiac fibrosis.Various studies have indicated that vitamin D displays an anti-fibrotic property in chroni...Objective Cardiac fibroblasts(CFs)proliferation and extracellular matrix deposition are important features of cardiac fibrosis.Various studies have indicated that vitamin D displays an anti-fibrotic property in chronic heart diseases.This study explored the role of vitamin D in the growth of CFs via an integrin signaling pathway.Methods MTT and 5-ethynyl-2′-deoxyuridine assays were performed to determine cell viability.Western blotting was performed to detect the expression of proliferating cell nuclear antigen(PCNA)and integrin signaling pathway.The fibronectin was observed by ELISA.Immunohistochemical staining was employed to evaluate the expression of integrinβ3.Results The PCNA expression in the CFs was enhanced after isoproterenol(ISO)stimulation accompanied by an elevated expression of integrin beta-3(β3).The blockade of the integrinβ3 with a specific integrinβ3 antibody reduced the PCNA expression induced by the ISO.Decreasing the integrinβ3 by siRNA reduced the ISO-triggered phosphorylation of FAK and Akt.Both the FAK inhibitor and Akt inhibitor suppressed the PCNA expression induced by the ISO in the CFs.Calcitriol(CAL),an active form of vitamin D,attenuated the ISO-induced CFs proliferation by downregulating the integrinβ3 expression,and phosphorylation of FAK and Akt.Moreover,CAL reduced the increased levels of fibronectin and hydroxyproline in the CFs culture medium triggered by the ISO.The administration of calcitriol decreased the integrinβ3 expression in the ISO-induced myocardial injury model.Conclusion These findings revealed a novel role for CAL in suppressing the CFs growth by the downregulation of the integrinβ3/FAK/Akt pathway.展开更多
[Objectives]To explore the effects of paeonol on the inhibition of myocardial fibrosis in high glucose induced cardiac fibroblasts(CFs).[Methods]Differential adherence method was used to culture the primary CFs of neo...[Objectives]To explore the effects of paeonol on the inhibition of myocardial fibrosis in high glucose induced cardiac fibroblasts(CFs).[Methods]Differential adherence method was used to culture the primary CFs of neonatal rats(passage culture);CCK-8 method was used to detect the cell proliferation;MTT assay was used to screen the safe concentration of paeonol;the 2 nd to 3 rd generation CFs were randomly divided into normal group(5.5 mmol/L,expressed in C),high glucose group(30 mmol/L,expressed in HG),paeonol low dose group(Pae-L,17.5 mg/L),and medium dose paeonol group(Pae-M,35 mg/L),paeonol high dose group(Pae-H,70 mg/L);Western Blot method was used to detect the expression of Col-I and Col-III protein.[Results]The extraction of CFs from primary neonatal rats was successful;high glucose(30 mmol/L)induction had a significant proliferation effect on CFs;compared with the normal group,the expressions of COI-I and Col-III protein were increased in the high glucose group(P<0.05);compared with the high glucose group,the expression of COI-I protein was decreased in each treatment group(P<0.05,P<0.01),and the decrease was most significant in the high dose group(P<0.01);the expression of COI-III in the high dose group was decreased and it was statistically significant(P<0.05).[Conclusions]Paeonol significantly inhibited the proliferation of high glucose induced CFs in neonatal rats.This experiment is intended to provide a new experimental basis for the prevention and treatment of diabetic cardiomyopathy(DCM).展开更多
Despite complications were significantly reduced due to the popularity of percutaneous coronary intervention(PCI) in clinical trials, reperfusion injury and chronic cardiac remodeling significantly contribute to poor ...Despite complications were significantly reduced due to the popularity of percutaneous coronary intervention(PCI) in clinical trials, reperfusion injury and chronic cardiac remodeling significantly contribute to poor prognosis and rehabilitation in AMI patients. We revealed the effects of HSP47 on myocardial ischemia-reperfusion injury(IRI) and shed light on the underlying molecular mechanism.We generated adult mice with lentivirus-mediated or miRNA(mi1/133TS)-aided cardiac fibroblastselective HSP47 overexpression. Myocardial IRI was induced by 45-min occlusion of the left anterior descending(LAD) artery followed by 24 h reperfusion in mice, while ischemia-mediated cardiac remodeling was induced by four weeks of reperfusion. Also, the role of HSP47 in fibrogenesis was evaluated in cardiac fibroblasts following hypoxia-reoxygenation(HR). Extensive HSP47 was observed in murine infarcted hearts, human ischemic hearts, and cardiac fibroblasts and accelerated oxidative stress and apoptosis after myocardial IRI. Cardiac fibroblast-selective HSP47 overexpression exacerbated cardiac dysfunction caused by chronic myocardial IRI and presented deteriorative fibrosis and cell proliferation.HSP47 upregulation in cardiac fibroblasts promoted TGFβ1-Smad4 pathway activation and Smad4 deubiquitination by recruiting ubiquitin-specific peptidase 10(USP10) in fibroblasts. However, cardiac fibroblast specific USP10 deficiency abolished HSP47-mediated fibrogenesis in hearts. Moreover, blockage of HSP47 with Col003 disturbed fibrogenesis in fibroblasts following HR. Altogether, cardiac fibroblast HSP47 aggravates fibrosis post-myocardial IRI by enhancing USP10-dependent Smad4 deubiquitination,which provided a potential strategy for myocardial IRI and cardiac remodeling.展开更多
Background Macrophage migration inhibitory factor （MIF） possesses proinflammatory function when secreted from the cells, and it also exhibits antioxidant properties based on its intrinsic oxidoreductase activity. Ho...Background Macrophage migration inhibitory factor （MIF） possesses proinflammatory function when secreted from the cells, and it also exhibits antioxidant properties based on its intrinsic oxidoreductase activity. However, the role of MIF in cardiac fibrosis is not well known. In the present study, the effect of MIF on fibrosis-associated gene expression and the underlying mechanism were examined. Methods The collagen content in mouse myocardium was detected by Masson staining. Expressions of MIF and fibrosis-associated Collal, Col3al and α-SMA in mouse myocardium or mouse cardiac fibroblasts were detected by quantitative real-time PCR and Western blot assay, respectively. Mature miR-29b expressions in mouse myocardium and cardiac fibroblasts were determined by real-time PCR. Smad3 activation in MIF-treated cardiac fibroblasts was also detected by Western blot assay. Results Compared with the db/m control mice, the collagen content was significantly increased in the myocardium of diabetic db/db mice. MIF was up-regulated, but miR-29b was down-regulated in the diabetic myocardium. Quantitative real-time PCR and Western blot assay showed that MIF could inhibit fibrosis-associated Collal, Col3al and α-SMA expressions in mouse cardiac fibroblasts. Smad3 activation was inhibited, but miR-29b was up-regulated in MIF-treated cardiac fibroblasts. Enforced expression of miR-29b significantly suppressed Collal, Col3al, and oL-SMA mRNA and lprotein expressions in cardiac fibroblasts. Conclusions MIF possesses the anti-fibrosis activity through inhibiting Smad3 activation and through up-regulating miR-29b expression, and miR-29b can inhibit fibrosis-associated Collal, Col3al and α-SMA expressions in cardiac fibroblasts.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81441016)and Key R&D Plan in Shaanxi Province of China(No.2020SF-262 and No.2019SF-200).
文摘Objective Cardiac fibroblasts(CFs)proliferation and extracellular matrix deposition are important features of cardiac fibrosis.Various studies have indicated that vitamin D displays an anti-fibrotic property in chronic heart diseases.This study explored the role of vitamin D in the growth of CFs via an integrin signaling pathway.Methods MTT and 5-ethynyl-2′-deoxyuridine assays were performed to determine cell viability.Western blotting was performed to detect the expression of proliferating cell nuclear antigen(PCNA)and integrin signaling pathway.The fibronectin was observed by ELISA.Immunohistochemical staining was employed to evaluate the expression of integrinβ3.Results The PCNA expression in the CFs was enhanced after isoproterenol(ISO)stimulation accompanied by an elevated expression of integrin beta-3(β3).The blockade of the integrinβ3 with a specific integrinβ3 antibody reduced the PCNA expression induced by the ISO.Decreasing the integrinβ3 by siRNA reduced the ISO-triggered phosphorylation of FAK and Akt.Both the FAK inhibitor and Akt inhibitor suppressed the PCNA expression induced by the ISO in the CFs.Calcitriol(CAL),an active form of vitamin D,attenuated the ISO-induced CFs proliferation by downregulating the integrinβ3 expression,and phosphorylation of FAK and Akt.Moreover,CAL reduced the increased levels of fibronectin and hydroxyproline in the CFs culture medium triggered by the ISO.The administration of calcitriol decreased the integrinβ3 expression in the ISO-induced myocardial injury model.Conclusion These findings revealed a novel role for CAL in suppressing the CFs growth by the downregulation of the integrinβ3/FAK/Akt pathway.
文摘[Objectives]To explore the effects of paeonol on the inhibition of myocardial fibrosis in high glucose induced cardiac fibroblasts(CFs).[Methods]Differential adherence method was used to culture the primary CFs of neonatal rats(passage culture);CCK-8 method was used to detect the cell proliferation;MTT assay was used to screen the safe concentration of paeonol;the 2 nd to 3 rd generation CFs were randomly divided into normal group(5.5 mmol/L,expressed in C),high glucose group(30 mmol/L,expressed in HG),paeonol low dose group(Pae-L,17.5 mg/L),and medium dose paeonol group(Pae-M,35 mg/L),paeonol high dose group(Pae-H,70 mg/L);Western Blot method was used to detect the expression of Col-I and Col-III protein.[Results]The extraction of CFs from primary neonatal rats was successful;high glucose(30 mmol/L)induction had a significant proliferation effect on CFs;compared with the normal group,the expressions of COI-I and Col-III protein were increased in the high glucose group(P<0.05);compared with the high glucose group,the expression of COI-I protein was decreased in each treatment group(P<0.05,P<0.01),and the decrease was most significant in the high dose group(P<0.01);the expression of COI-III in the high dose group was decreased and it was statistically significant(P<0.05).[Conclusions]Paeonol significantly inhibited the proliferation of high glucose induced CFs in neonatal rats.This experiment is intended to provide a new experimental basis for the prevention and treatment of diabetic cardiomyopathy(DCM).
基金supported by grants from the National Key R&D Program of China (2018YFC1311300)the Key Project of the National Natural Science Foundation of China (No. 81530012)+3 种基金the National Natural Science Foundation of China (Nos. 81860080, 82170245 and 81700254)the Fundamental Research Funds for the Central Universities (2042018kf1032, China)the Development Center for Medical Science and Technology National Health and Family Planning Commission of the People’s Republic of China (The prevention and control project of cardiovascular disease, 2016ZX008-01)Science and Technology Planning Projects of Wuhan (2018061005132295)
文摘Despite complications were significantly reduced due to the popularity of percutaneous coronary intervention(PCI) in clinical trials, reperfusion injury and chronic cardiac remodeling significantly contribute to poor prognosis and rehabilitation in AMI patients. We revealed the effects of HSP47 on myocardial ischemia-reperfusion injury(IRI) and shed light on the underlying molecular mechanism.We generated adult mice with lentivirus-mediated or miRNA(mi1/133TS)-aided cardiac fibroblastselective HSP47 overexpression. Myocardial IRI was induced by 45-min occlusion of the left anterior descending(LAD) artery followed by 24 h reperfusion in mice, while ischemia-mediated cardiac remodeling was induced by four weeks of reperfusion. Also, the role of HSP47 in fibrogenesis was evaluated in cardiac fibroblasts following hypoxia-reoxygenation(HR). Extensive HSP47 was observed in murine infarcted hearts, human ischemic hearts, and cardiac fibroblasts and accelerated oxidative stress and apoptosis after myocardial IRI. Cardiac fibroblast-selective HSP47 overexpression exacerbated cardiac dysfunction caused by chronic myocardial IRI and presented deteriorative fibrosis and cell proliferation.HSP47 upregulation in cardiac fibroblasts promoted TGFβ1-Smad4 pathway activation and Smad4 deubiquitination by recruiting ubiquitin-specific peptidase 10(USP10) in fibroblasts. However, cardiac fibroblast specific USP10 deficiency abolished HSP47-mediated fibrogenesis in hearts. Moreover, blockage of HSP47 with Col003 disturbed fibrogenesis in fibroblasts following HR. Altogether, cardiac fibroblast HSP47 aggravates fibrosis post-myocardial IRI by enhancing USP10-dependent Smad4 deubiquitination,which provided a potential strategy for myocardial IRI and cardiac remodeling.
基金supported by Grants from the National Natural Science Foundation of China(No.81070102/81270222/81273516/81302779)Foundation of the Natural Science Foundation of the Guangdong Province(No.S2011020005911/S2012010009453)
文摘Background Macrophage migration inhibitory factor （MIF） possesses proinflammatory function when secreted from the cells, and it also exhibits antioxidant properties based on its intrinsic oxidoreductase activity. However, the role of MIF in cardiac fibrosis is not well known. In the present study, the effect of MIF on fibrosis-associated gene expression and the underlying mechanism were examined. Methods The collagen content in mouse myocardium was detected by Masson staining. Expressions of MIF and fibrosis-associated Collal, Col3al and α-SMA in mouse myocardium or mouse cardiac fibroblasts were detected by quantitative real-time PCR and Western blot assay, respectively. Mature miR-29b expressions in mouse myocardium and cardiac fibroblasts were determined by real-time PCR. Smad3 activation in MIF-treated cardiac fibroblasts was also detected by Western blot assay. Results Compared with the db/m control mice, the collagen content was significantly increased in the myocardium of diabetic db/db mice. MIF was up-regulated, but miR-29b was down-regulated in the diabetic myocardium. Quantitative real-time PCR and Western blot assay showed that MIF could inhibit fibrosis-associated Collal, Col3al and α-SMA expressions in mouse cardiac fibroblasts. Smad3 activation was inhibited, but miR-29b was up-regulated in MIF-treated cardiac fibroblasts. Enforced expression of miR-29b significantly suppressed Collal, Col3al, and oL-SMA mRNA and lprotein expressions in cardiac fibroblasts. Conclusions MIF possesses the anti-fibrosis activity through inhibiting Smad3 activation and through up-regulating miR-29b expression, and miR-29b can inhibit fibrosis-associated Collal, Col3al and α-SMA expressions in cardiac fibroblasts.
