Electroacupuncture improves myocardial fibrosis in heart failure rats by attenuating ECM collagen deposition through modulation of TGF-β1/Smads signaling pathway

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.

蒿鳖养阴软坚方对TGF-β1诱导的肝纤维化的作用

目的:研究蒿鳖养阴软坚方在体外对LX-2细胞的影响,从而探讨蒿鳖养阴软坚方对TGF-β1诱导的肝纤维化的作用。方法:体外培养人肝星状细胞LX-2,实验分为正常对照组、TGF-β1刺激组和蒿鳖养阴软坚方低、中、高浓度给药组。MTT法检测LX-2细胞增殖情况;比色法检测细胞培养液中乳酸脱氢酶(LDH)的活性;酶消化法检测细胞上清羟脯氨酸(Hyp)的含量;Western blot法检测collagenⅠ以及α-SMA蛋白表达量。结果:TGF-β1作用于LX-2能够明显促进细胞的增殖,collagen Ⅰ、α-SMA的表达量显著升高。蒿鳖养阴软坚方能抑制LX-2细胞增殖并且能够降低collagenⅠ和α-SMA的表达。结论:蒿鳖养阴软坚方能够显著抑制LX-2增殖,下调collagenⅠ和α-SMA的表达,降低Hyp生成,从而发挥抗肝纤维化的作用。

喘可治注射液对人胚胎肺成纤维细胞转化因子及相关蛋白的影响

目的:通过喘可治注射液(CKZ)干预人胚胎肺成纤维细胞(Wi-38),测定β转化生长因子(TGF-β)、1型胶原纤维蛋白(collagen1)和α-平滑肌肌动蛋白(α-SMA),探讨CKZ在抑制细胞间质转化方面的作用机制。方法:将体外培养的Wi-38细胞随机分为6组:① 对照组,单独的DMEM培养;② 5 ng/mL TGF-β组,加入浓度为5 ng/mL的TGF-β孵育;③ 10 ng/mL TGF-β组,加入浓度为10 ng/mL的TGF-β孵育;④10 ul/mL喘可治注射液(CKZ)组,加入浓度为10 ul/mL CKZ的孵育;⑤ 20 ul/mL CKZ组,加入浓度为20 ul/mL CKZ的孵育;⑥ 10 ng/mL TGF-β + 20 ul/mL CKZ组,同时加入浓度10 ng/mL TGF-β和20 ul/mL CKZ孵育。采用实时荧光定量(qPCR)的方法检测孵育24 h、48 h、72 h的TGF-β、collagen 1、α-SMA的表达水平变化。结果:加入TGF-β处理组细胞的α-SMA、Collagen 1的mRNA水平升高、TGF-β的mRNA水平降低,与对照组相比,其差异均具有统计学意义(P 【0.05);加入CKZ处理组细胞的α-SMA、TGF-β、Collagen 1的mRNA水平均降低,与对照组相比,其差异具有统计学意义(P 【0.05);同时加入TGF-β和CKZ处理组细胞的α-SMA、Collagen 1的mRNA水平均显著降低,与TGF-β处理组相比,其差异具有统计学意义(P 【0.05)。结论:TGF-β、collagen 1、α-SMA参与了人胚胎肺成纤维细胞间质转化的病理进程,CKZ对这一进程起到了抑制作用,CKZ具有一定的抑制气道上皮间质转化的作用。

大王马先蒿对四氯化碳诱导肝纤维化小鼠的保护作用

目的探讨大王马先蒿对肝纤维化的保护作用。方法将昆明小鼠随机分为正常对照组、模型对照组、秋水仙碱(0.1 mg·kg^(-1))组及大王马先蒿小(200 mg·kg^(-1))、中(400 mg·kg^(-1))、大(800 mg·kg^(-1))剂量组,每组10只;通过腹腔注射四氯化碳(CCl_(4),1 mL·kg^(-1))建立肝纤维化模型,并于第5周开始灌胃给药,每天1次,持续至第12周结束;末次给药后1 h,麻醉眼球取血检测血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)及IV型胶原(Ⅳ-C)水平,处死小鼠并取出脏器,计算小鼠脏器系数,Western blotting检测肝组织TGF-β_(1),Smad3,α-SMA及Collagen Ⅰ表达,苏木精-伊红(HE)和马松(Masson)染色观察小鼠肝组织病理形态。结果模型对照组小鼠皮毛竖立,活动减少,反应迟缓,大王马先蒿组小鼠一般状态好于模型对照组,反应较敏捷,皮色白有光泽。与模型对照组比较,大王马先蒿各剂量均能显著改善CCl_(4)所致小鼠肝组织的肝纤维化程度,显著降低血清ALT、AST、HA、LN、PCⅢ及Ⅳ-C水平(P<0.01),显著下调TGF-β_(1),Smad3,α-SMA及Collagen Ⅰ蛋白表达(P<0.01)。结论大王马先蒿对CCl_(4)诱导的肝纤维化小鼠具有较好的保护作用,该作用可能与抑制TGF-β_(1)/Smad信号通路及下调α-SMA和Collagen Ⅰ蛋白的表达有关。