We studied the effects and mechanisms of action of conophylline in different concentrations in the original in vitro model of myocardial fibrosis (treatment of cardiac fibroblasts isolated form the hearts of newborn rats with angiotensin II). Viability, collagen content, and expression of related protein in cardiac fibroblasts were assessed using the MTT-test, Sircol assay, and Western blotting, respectively. Conophylline markedly protected the cultured cells against the development of angiotensin II-induced fibrosis, which was seen from reduced viability of fibroblasts, decreased collagen content, and down-regulation of the expression of α-smooth muscle actin (α-SMA). Conophylline did not affect the TGF-β pathway altered by angiotensin II, but markedly decreased the level of bone morphogenetic protein-4 (BMP4) enhanced by angiotensin II and BMP4 itself. Conophylline produced no effect on phosphorylation of α-SMA and Smad homologue-1/5/8, the classic BMP4 downstream pathway elements, but reduced the level of c-Jun N-terminal kinase (JNK) elevated by BMP4. Conophylline did not inhibit the development of myocardial fibrosis in the presence of JNK activator anisomycin. Thus, conophylline inhibited angiotensin II-provoked myocardial fibrosis via the BMP4/JNK pathway.

我们在体外研究了不同浓度的茶碱的作用和作用机制。心肌纤维化模型（用血管紧张素 II 治疗从新生大鼠心脏中分离的心脏成纤维细胞）。分别使用 MTT 检验、Sircol 测定和蛋白质印迹法评估心脏成纤维细胞中的活力、胶原含量和相关蛋白的表达。芋螺碱显着保护培养的细胞免受血管紧张素 II 诱导的纤维化的发展，这可以从成纤维细胞活力降低、胶原蛋白含量降低和 α-平滑肌肌动蛋白 (α-SMA) 表达下调中看出。芋螺碱不影响血管紧张素 II 改变的 TGF-β 通路，但显着降低血管紧张素 II 和 BMP4 本身增强的骨形态发生蛋白 4 (BMP4) 的水平。芋螺碱对 α-SMA 和 Smad 同源物 1/5/8 的磷酸化没有影响，经典的 BMP4 下游途径元件，但降低了 BMP4 升高的 c-Jun N-末端激酶 (JNK) 的水平。在 JNK 激活剂茴香霉素存在的情况下，芋螺碱不抑制心肌纤维化的发展。因此，刀豆碱通过 BMP4/JNK 通路抑制血管紧张素 II 诱发的心肌纤维化。