imbalance of synovial macrophages in patients with rheumatoid arthritis(RA).Methods:Human mononuclear cells(THP‑1)at logarithmic growth stage were induced into M1‑type macrophages,and RA synovial fibroblasts M1‑type m...imbalance of synovial macrophages in patients with rheumatoid arthritis(RA).Methods:Human mononuclear cells(THP‑1)at logarithmic growth stage were induced into M1‑type macrophages,and RA synovial fibroblasts M1‑type macrophages were co‑cultured into synovial macrophages.Synovial macrophages were divided into four groups:RA group(blank group),TGF‑β1 group(model group)and miR145‑5P overexpression group(TGF‑β1+miR145‑5P mimics group)and miR145‑5P overexpression negative control group(TGF‑β1+miR145‑5P‑mimics‑NC group).The blank group did not receive any treatment,and the other three groups were induced by TGF‑β1 in the medium for 48 h.Transfection miR145‑5p mimic and miR145‑5P‑mimics‑NC were added to co‑culture medium,and IL‑6,IL‑6 and IL‑6 of synovial macrophages were detected by ELISA.CD163 expression.Rt‑qpcr was used to detect miR145‑5p mRNA,TGF‑β1mRNA,Smad3mRNA,Smad7mRNA expression level.The expression of TGF‑β1/Smads pathway related proteins was detected by Western Blotting.Results:Compared with blank group,IL‑6 level was up‑regulated(P<0.01),and CD163 level was down‑regulated in model group(P<0.05),suggesting that TGF‑β1 could induce intensified immune inflammatory response.Compared with the negative miR145‑5P overexpression control group and model group,The expression of miR145‑5P overexpression group molecule CD163 was significantly increased by ELISA(P<0.01),and the expression of inflammatory factor IL‑6 was decreased(P<0.05).PCR showed that miR145‑5P mRNA expression level was significantly increased in miR145‑5P overexpression group,Smad3 mRNA and TGF‑β1 mRNA were significantly decreased,and Smad7 mRNA was significantly increased(P<0.01).WB method showed that the anti‑inflammatory protein Smad7 was significantly increased,while TGF‑β1 and Smad3 were significantly decreased(P<0.01).Transwell chamber results confirmed that miR145‑5P overexpression group significantly reduced macrophage invasion(P<0.01).Correlation analysis showed that miR145‑5P was negatively correlated with Smad3 and positively correlated with Smad7(P<0.01).Conclusion:miR145‑5P may inhibit macrophage polarization in RA patients by targeting Smad3 protein,negatively regulating TGF‑β1/Smads pathway,and alleviating immune inflammation.展开更多
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
基金Natural Science Research Project of Colleges and Universities in Anhui Province(No.kj2020a0396)。
文摘imbalance of synovial macrophages in patients with rheumatoid arthritis(RA).Methods:Human mononuclear cells(THP‑1)at logarithmic growth stage were induced into M1‑type macrophages,and RA synovial fibroblasts M1‑type macrophages were co‑cultured into synovial macrophages.Synovial macrophages were divided into four groups:RA group(blank group),TGF‑β1 group(model group)and miR145‑5P overexpression group(TGF‑β1+miR145‑5P mimics group)and miR145‑5P overexpression negative control group(TGF‑β1+miR145‑5P‑mimics‑NC group).The blank group did not receive any treatment,and the other three groups were induced by TGF‑β1 in the medium for 48 h.Transfection miR145‑5p mimic and miR145‑5P‑mimics‑NC were added to co‑culture medium,and IL‑6,IL‑6 and IL‑6 of synovial macrophages were detected by ELISA.CD163 expression.Rt‑qpcr was used to detect miR145‑5p mRNA,TGF‑β1mRNA,Smad3mRNA,Smad7mRNA expression level.The expression of TGF‑β1/Smads pathway related proteins was detected by Western Blotting.Results:Compared with blank group,IL‑6 level was up‑regulated(P<0.01),and CD163 level was down‑regulated in model group(P<0.05),suggesting that TGF‑β1 could induce intensified immune inflammatory response.Compared with the negative miR145‑5P overexpression control group and model group,The expression of miR145‑5P overexpression group molecule CD163 was significantly increased by ELISA(P<0.01),and the expression of inflammatory factor IL‑6 was decreased(P<0.05).PCR showed that miR145‑5P mRNA expression level was significantly increased in miR145‑5P overexpression group,Smad3 mRNA and TGF‑β1 mRNA were significantly decreased,and Smad7 mRNA was significantly increased(P<0.01).WB method showed that the anti‑inflammatory protein Smad7 was significantly increased,while TGF‑β1 and Smad3 were significantly decreased(P<0.01).Transwell chamber results confirmed that miR145‑5P overexpression group significantly reduced macrophage invasion(P<0.01).Correlation analysis showed that miR145‑5P was negatively correlated with Smad3 and positively correlated with Smad7(P<0.01).Conclusion:miR145‑5P may inhibit macrophage polarization in RA patients by targeting Smad3 protein,negatively regulating TGF‑β1/Smads pathway,and alleviating immune inflammation.
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.
