Background: To explore the effects of electroacupuncture on cardiac function and myocardial fibrosis in rat models of heart failure, and to elucidate the underlying mechanism of electroacupuncture in heart failure tre...Background: To explore the effects of electroacupuncture on cardiac function and myocardial fibrosis in rat models of heart failure, and to elucidate the underlying mechanism of electroacupuncture in heart failure treatment. Methods: Healthy male Sprague-Dawley rats were allocated into three groups: Sham group, Model group, and electroacupuncture (Model + EA) group, with each group comprising 8 rats. The model underwent a procedure involving the ligation of the left anterior descending coronary artery to induce a model of heart failure. The Model + EA group was used for 7 consecutive days for electroacupuncture of bilateral Shenmen (HT7) and Tongli (HT5), once a day for 30 min each time. Left ventricular parameters in rats were assessed using a small-animal ultrasound machine to analyze changes in left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular ejection fraction, and left ventricular fractional shortening. Serum interleukin-1β (IL-1β), cardiac troponin (cTn), and N-terminal brain natriuretic peptide precursor levels were measured using ELISA. Histopathological changes in rat myocardium were observed through HE staining, while collagen deposition in rat myocardial tissue was assessed using the Masson staining method. Picro sirius red staining, immunohistochemical staining, and RT-qPCR were utilized to distinguish between the various types of collagen deposition. The expression level of TGF-β1 and SMAD2/3/4/7 mRNA in rat myocardial tissues was determined using RT-qPCR. Additionally, western blot analysis was conducted to assess the protein expression levels of TGF-β1, SMAD3/7, and p-SMAD3 in rat myocardial tissues. Results: Compared with the Sham group, the left ventricular ejection fraction and left ventricular fractional shortening values of the Model group were significantly decreased (P < 0.01);the left ventricular end-diastolic volume and left ventricular end-systolic volume values were remarkably increased (P < 0.01);serum N-terminal brain natriuretic peptide precursor content was increased (P < 0.01);serum IL-1β and cTn levels were increased (P < 0.01);myocardial collagen volume fraction were increased (P < 0.01);and those of the expression of TGF-β1 and SMAD2/3/4 mRNA was increased (P < 0.01);the expression of SMAD7 mRNA was decreased (P < 0.01);the protein expression levels of TGF-β1, SMAD3, and p-Smad3 were increased (P < 0.01);the protein expression level of SMAD7 was decreased (P < 0.01) in the Model group. Compared to the Model group, the expression levels of the proteins TGF-β1, SMAD3, and p-Smad3 in myocardial tissue were found to be decreased (P < 0.01), and the expression level of the protein SMAD7 was found to be increased (P < 0.01) in the Model + EA group;the collagen volume fraction and deposition of type Ⅰ /Ⅲ collagen were decreased (P < 0.01) in the Model + EA group. Conclusion: Electroacupuncture alleviates myocardial fibrosis in rats with heart failure, and this effect is likely due to attributed to the modulation of the TGF-β1/Smads signaling pathway, which helps reduce collagen deposition in the extracellular matrix.展开更多
The purpose of the present study was to study the impacts of eplerenone （EPL）, an antagonist of mineralocorticoid receptors （MR）, on atrial fibrosis in a mouse model with selective fibrosis in the atrium, and to e...The purpose of the present study was to study the impacts of eplerenone （EPL）, an antagonist of mineralocorticoid receptors （MR）, on atrial fibrosis in a mouse model with selective fibrosis in the atrium, and to explore the possible mechanisms. Using mutant TGF-β1 transgenic （Tx） mice, we first demonstrated that EPL inhibited atrial fibrosis specifically and decreased mac- rophage accumulation in the atria of these mice. Results from immunohistochemistry and western blotting showed that EPL attenuated protein expression of fibrosis-related molecules such as connective tissue growth factor （CTGF） and fibronectin in the atria of Tx mice. In culture, EPL inhibited gene expression of fibrosis-related molecules such as fibronectin, ct-SMA, and CTGF in TGF-β1-stimulated atrial fibroblasts, Finally, using a co-culture system, we showed that TGF-β1 stimulated atrial fi- broblasts induced migration of macrophages and this was blocked by EPL. EPL also blocked TGF-β1 induced gene expression of intedeukin-6 （IL-6） in atrial fibroblasts. Therefore, we conclude that EPL attenuated atrial fibrosis and macrophage infiltra- tion in Tx mice. TGF-I31 and IL-6 were involved in the impacts of EPL on activation of atrial fibroblasts and interactions be- tween fibroblasts and macrophages.展开更多
Background Myocardial fibrosis plays a critical role in the process of diabetic cardiac remolding.MicroRNAs （miRNAs） are endogenous,small non-coding RNAs that negatively regulate gene expression in diverse biologica...Background Myocardial fibrosis plays a critical role in the process of diabetic cardiac remolding.MicroRNAs （miRNAs） are endogenous,small non-coding RNAs that negatively regulate gene expression in diverse biological and pathological processes.However,the roles of miRNAs in myocardial fibrosis have not been well elucidated.In the present study,miRNAs profiles in the fibrotic myocardium of db/db mice and miRNAs expression in TGF-β1-stimulated mouse cardiac myofibroblasts was examined.Methods Heart function of 18-week-old db/db mice and db/m control mice was detected by echocardiography.miRNA expression profile in diabetic myocardium was detected by miRNA microarray.Quantitative real-time PCR was used to determine the expression of fibrosis-related genes and miRNA precursors of interest.Western blot was used to detect the levels of fibrosis-related proteins,activated Smad3 and total Smad3.Results The result of echocardiography showed that left ventricular systolic and diastolic function was impaired in 18-week-old db/db mice without significant change of ejection fraction （EF） and fractional shortening （FS）.Fibrosis-related genes expression was upregulated and the amount of phosphorylated Smad3 was increased significantly in the diabetic myocardium.miRNAs dysregulation was shown in diabetic myocardium,sixty-eight miRNAs,including miR-208b,miR-29b,miR-26b and miR-30e,were increased over two-fold,meanwhile,sixty-two miRNAs were decreased more than two-fold in the myocardium of db/db mice compared to db/m controls.In parallel with a significant upregulation of Col1a1,Col3a1 and CTGF miRNA expression,miR-208b,miR-29b,miR-26b and miR-30e precursors were also shown to be upregulated in TGF-β1-induced C57bl/6 mouse cardiac myofibroblasts.Conclusions microRNAs were dysregulated in diabetic myocardium,with the activation of TGF-β/smad3 pathway,contributing to diabetic myocardial fibrosis.展开更多
基金the China’s National Key Research and Development Program Projects(No.2022YFC3500500 and No.2022YFC3500502).
文摘Background: To explore the effects of electroacupuncture on cardiac function and myocardial fibrosis in rat models of heart failure, and to elucidate the underlying mechanism of electroacupuncture in heart failure treatment. Methods: Healthy male Sprague-Dawley rats were allocated into three groups: Sham group, Model group, and electroacupuncture (Model + EA) group, with each group comprising 8 rats. The model underwent a procedure involving the ligation of the left anterior descending coronary artery to induce a model of heart failure. The Model + EA group was used for 7 consecutive days for electroacupuncture of bilateral Shenmen (HT7) and Tongli (HT5), once a day for 30 min each time. Left ventricular parameters in rats were assessed using a small-animal ultrasound machine to analyze changes in left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular ejection fraction, and left ventricular fractional shortening. Serum interleukin-1β (IL-1β), cardiac troponin (cTn), and N-terminal brain natriuretic peptide precursor levels were measured using ELISA. Histopathological changes in rat myocardium were observed through HE staining, while collagen deposition in rat myocardial tissue was assessed using the Masson staining method. Picro sirius red staining, immunohistochemical staining, and RT-qPCR were utilized to distinguish between the various types of collagen deposition. The expression level of TGF-β1 and SMAD2/3/4/7 mRNA in rat myocardial tissues was determined using RT-qPCR. Additionally, western blot analysis was conducted to assess the protein expression levels of TGF-β1, SMAD3/7, and p-SMAD3 in rat myocardial tissues. Results: Compared with the Sham group, the left ventricular ejection fraction and left ventricular fractional shortening values of the Model group were significantly decreased (P < 0.01);the left ventricular end-diastolic volume and left ventricular end-systolic volume values were remarkably increased (P < 0.01);serum N-terminal brain natriuretic peptide precursor content was increased (P < 0.01);serum IL-1β and cTn levels were increased (P < 0.01);myocardial collagen volume fraction were increased (P < 0.01);and those of the expression of TGF-β1 and SMAD2/3/4 mRNA was increased (P < 0.01);the expression of SMAD7 mRNA was decreased (P < 0.01);the protein expression levels of TGF-β1, SMAD3, and p-Smad3 were increased (P < 0.01);the protein expression level of SMAD7 was decreased (P < 0.01) in the Model group. Compared to the Model group, the expression levels of the proteins TGF-β1, SMAD3, and p-Smad3 in myocardial tissue were found to be decreased (P < 0.01), and the expression level of the protein SMAD7 was found to be increased (P < 0.01) in the Model + EA group;the collagen volume fraction and deposition of type Ⅰ /Ⅲ collagen were decreased (P < 0.01) in the Model + EA group. Conclusion: Electroacupuncture alleviates myocardial fibrosis in rats with heart failure, and this effect is likely due to attributed to the modulation of the TGF-β1/Smads signaling pathway, which helps reduce collagen deposition in the extracellular matrix.
基金supported by National Nature Science Foundation of China(30871083)Doctoral Innovation Fund Projects from Shanghai Jiao Tong University School of Medicine(BXJ201442)
文摘The purpose of the present study was to study the impacts of eplerenone （EPL）, an antagonist of mineralocorticoid receptors （MR）, on atrial fibrosis in a mouse model with selective fibrosis in the atrium, and to explore the possible mechanisms. Using mutant TGF-β1 transgenic （Tx） mice, we first demonstrated that EPL inhibited atrial fibrosis specifically and decreased mac- rophage accumulation in the atria of these mice. Results from immunohistochemistry and western blotting showed that EPL attenuated protein expression of fibrosis-related molecules such as connective tissue growth factor （CTGF） and fibronectin in the atria of Tx mice. In culture, EPL inhibited gene expression of fibrosis-related molecules such as fibronectin, ct-SMA, and CTGF in TGF-β1-stimulated atrial fibroblasts, Finally, using a co-culture system, we showed that TGF-β1 stimulated atrial fi- broblasts induced migration of macrophages and this was blocked by EPL. EPL also blocked TGF-β1 induced gene expression of intedeukin-6 （IL-6） in atrial fibroblasts. Therefore, we conclude that EPL attenuated atrial fibrosis and macrophage infiltra- tion in Tx mice. TGF-I31 and IL-6 were involved in the impacts of EPL on activation of atrial fibroblasts and interactions be- tween fibroblasts and macrophages.
基金supported by Grants from the National Natural Science Foundation of China (No.81070102)the Natural Science Foundation of the Guangdong Province (No.10151008004000035,S2011020005911)
文摘Background Myocardial fibrosis plays a critical role in the process of diabetic cardiac remolding.MicroRNAs （miRNAs） are endogenous,small non-coding RNAs that negatively regulate gene expression in diverse biological and pathological processes.However,the roles of miRNAs in myocardial fibrosis have not been well elucidated.In the present study,miRNAs profiles in the fibrotic myocardium of db/db mice and miRNAs expression in TGF-β1-stimulated mouse cardiac myofibroblasts was examined.Methods Heart function of 18-week-old db/db mice and db/m control mice was detected by echocardiography.miRNA expression profile in diabetic myocardium was detected by miRNA microarray.Quantitative real-time PCR was used to determine the expression of fibrosis-related genes and miRNA precursors of interest.Western blot was used to detect the levels of fibrosis-related proteins,activated Smad3 and total Smad3.Results The result of echocardiography showed that left ventricular systolic and diastolic function was impaired in 18-week-old db/db mice without significant change of ejection fraction （EF） and fractional shortening （FS）.Fibrosis-related genes expression was upregulated and the amount of phosphorylated Smad3 was increased significantly in the diabetic myocardium.miRNAs dysregulation was shown in diabetic myocardium,sixty-eight miRNAs,including miR-208b,miR-29b,miR-26b and miR-30e,were increased over two-fold,meanwhile,sixty-two miRNAs were decreased more than two-fold in the myocardium of db/db mice compared to db/m controls.In parallel with a significant upregulation of Col1a1,Col3a1 and CTGF miRNA expression,miR-208b,miR-29b,miR-26b and miR-30e precursors were also shown to be upregulated in TGF-β1-induced C57bl/6 mouse cardiac myofibroblasts.Conclusions microRNAs were dysregulated in diabetic myocardium,with the activation of TGF-β/smad3 pathway,contributing to diabetic myocardial fibrosis.
