Liu Menghui, Lin Yue, Xu Huibo, Li Lixin, Ding Tao
Department of Traditional Chinese Medicine, Changchun University of Traditional Chinese Medicine, Changchun, Jilin 130000, P.R. China.
Department of Pediatrics, Jilin Academy of Traditional Chinese Medicine, Changchun, Jilin 130000, P.R. China.
Exp Ther Med. 2022 Jul 14;24(3):570. doi: 10.3892/etm.2022.11507. eCollection 2022 Sep.
Chronic cardiac inflammation and fibrosis can progress into severe forms of cardiomyopathy. alkaloids (KuShen) have been previously reported to exert anti-inflammatory effects, whereas saponins (XiYangShen) has been shown to alleviate cardiac fibrosis. Therefore, the potential effects of their combination (KX) on different stages of autoimmune myocarditis were investigated in the present study. Mice were randomly divided into the following four groups: Control; experimental autoimmune myocarditis (EAM); KX-High (275 mg/kg); and KX-Low (138 mg/kg). A 21-day and a 60-day EAM model was established through multi-site subcutaneous injections of cardiac myosin mixed with complete Freund's adjuvant on days 0, 7, 21 and 42. Mice in the High and Low KX groups were treated by gavage (10 ml/kg) daily from day 0 (1 day before treatment) until sacrifice (day 21 or 60). Mice in the control and EAM groups received an equivalent volume of distilled water. The levels of lactate dehydrogenase (LDH), creatine kinase-myocardial band (CK-MB), cardiac troponin I (cTn-I), IL-1β, IL-6, TNF-α, TGF-β1, collagen type I (Col Ⅰ) and collagen type III (Col Ⅲ) were measured by ELISA in the mouse myocardial tissues or serum. Myocardial tissue structure and extent of fibrosis were visualized using H&E and Masson's staining. Western blotting and immunohistochemistry were used to measure the expression levels NF-κB and TGF-β1 pathway proteins in the myocardial tissues. The degree of inflammation in the 21-day EAM model was found to be significantly higher compared with that in the 60-day EAM model. KX significantly reduced the inflammatory response at 21 days by decreasing the expression levels of CK-MB, LDH, cTn-I, IL-1β, IL-6, TNF-α and TGF-β-activated kinase 1-binding protein 1/NF-κB pathway proteins. Myocardial fibrosis in the 60-day EAM model was also significantly worse compared with that in the 21-day EAM model. However, fibrosis was significantly delayed by treatment with KX. In addition, KX significantly decreased the expression levels of TGF-β1, Smad2, Smad4, Col I and Col III. Therefore, these data suggest that KX is beneficial for treating myocarditis by targeting multiple pathways.
慢性心脏炎症和纤维化可进展为严重形式的心肌病。此前有报道称生物碱（苦参）具有抗炎作用，而皂苷（西洋参）已被证明可减轻心脏纤维化。因此，本研究探讨了它们的组合（KX）对自身免疫性心肌炎不同阶段的潜在影响。将小鼠随机分为以下四组：对照组；实验性自身免疫性心肌炎（EAM）组；KX高剂量组（275毫克/千克）；KX低剂量组（138毫克/千克）。通过在第0、7、21和42天多点皮下注射心肌肌凝蛋白与完全弗氏佐剂混合液建立21天和60天的EAM模型。KX高剂量组和低剂量组的小鼠从第0天（治疗前1天）开始每天通过灌胃（10毫升/千克）给药，直至处死（第21天或60天）。对照组和EAM组的小鼠接受等量的蒸馏水。通过酶联免疫吸附测定法（ELISA）检测小鼠心肌组织或血清中乳酸脱氢酶（LDH）、肌酸激酶同工酶（CK-MB）、心肌肌钙蛋白I（cTn-I）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、转化生长因子-β1（TGF-β1）、I型胶原（Col Ⅰ）和III型胶原（Col Ⅲ）的水平。使用苏木精-伊红（H&E）染色和Masson染色观察心肌组织结构和纤维化程度。采用蛋白质印迹法和免疫组织化学法检测心肌组织中核因子-κB（NF-κB）和TGF-β1信号通路蛋白的表达水平。发现21天EAM模型中的炎症程度明显高于60天EAM模型。KX通过降低CK-MB、LDH、cTn-I、IL-1β、IL-6、TNF-α和TGF激活激酶1结合蛋白1/NF-κB信号通路蛋白的表达水平，在21天时显著减轻炎症反应。60天EAM模型中的心肌纤维化也明显比21天EAM模型更严重。然而，KX治疗可显著延缓纤维化进程。此外，KX显著降低了TGF-β1、Smad2、Smad4、Col I和Col III的表达水平。因此，这些数据表明KX通过靶向多种信号通路对治疗心肌炎有益。
