Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, 350122, China; Chen Keji Academic Thought Inheritance Studio, Fujian University of Traditional Chinese Medicine, Fuzhou, 350122, China; Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou, 350122, China.
The Third People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine, Fuzhou, 350122, China.
J Ethnopharmacol. 2021 Jul 15;275:114061. doi: 10.1016/j.jep.2021.114061. Epub 2021 Apr 21.
The abnormal proliferation and differentiation of cardiac fibroblasts (CFs) are universally regarded as the key process for the progressive development of cardiac fibrosis following various cardiovascular diseases. Huoxin Pill (Concentrated pill, HXP) is a Chinese herbal formula for treating coronary heart disease. However, the cellular and molecular mechanisms of HXP in the treatment of myocardial fibrosis are still unclear.
To investigate the effects of HXP on CFs transdifferentiation and collagen synthesis under isoproterenol (ISO) conditions, as well as the potential mechanism of action.
In vivo, we established a rat model of cardiac fibrosis induced by ISO, and administered with low or high dose of HXP (10 mg/kg/day or 30 mg/kg/day). The level of α-SMA was detected by immunohistochemistry examination, and combined with RNA-sequencing analysis to determine the protective effect of HXP on myocardial fibrosis rats. In vitro, by culturing primary rat CFs, we examined the effects of HXP on the proliferation and transdifferentiation of CFs using CCK8, scratch wound healing and immunofluorescence assays. Western blot was used to determine protein expression.
The findings revealed that HXP protects against ISO-induced cardiac fibrosis and CFs transdifferentiation in rats. RNA-sequencing and pathway analyses demonstrated 238 or 295 differentially expressed genes (DEGs) and multiple enriched signal pathways, including transforming growth factor-beta (TGF-β) receptor signaling activates Smads, downregulation of TGF-β receptor signaling, signaling by TGF-β receptor complex, and collagen formation under treatment with low or high-dose of HXP. Moreover, HXP also markedly inhibited ISO-induced primary rat CFs proliferation, transdifferentiation, collagen synthesis and the upregulation of TGF-β1 and phosphorylated Smad2/3 protein expression.
HXP suppresses ISO-induced CFs transdifferentiation and collagen synthesis, and it may exert these effects in part by inhibiting the activation of the TGF-β/Smads pathway. This may be a new therapeutic tool for cardiac fibrosis.
心脏成纤维细胞（CFs）的异常增殖和分化被普遍认为是各种心血管疾病后心脏纤维化进行性发展的关键过程。活心丸（浓缩丸，HXP）是一种治疗冠心病的中药方剂。然而，HXP 治疗心肌纤维化的细胞和分子机制尚不清楚。
研究 HXP 在异丙肾上腺素（ISO）条件下对 CFs 转分化和胶原合成的影响，以及潜在的作用机制。
体内，我们建立了 ISO 诱导的大鼠心肌纤维化模型，并给予低或高剂量 HXP（10mg/kg/天或 30mg/kg/天）。通过免疫组织化学检查检测α-SMA 的水平，并结合 RNA 测序分析确定 HXP 对心肌纤维化大鼠的保护作用。体外，通过培养原代大鼠 CFs，我们使用 CCK8、划痕愈合和免疫荧光测定法研究 HXP 对 CFs 增殖和转分化的影响。使用 Western blot 测定蛋白表达。
研究结果表明，HXP 可预防 ISO 诱导的大鼠心脏纤维化和 CFs 转分化。RNA 测序和通路分析显示，低或高剂量 HXP 处理后有 238 个或 295 个差异表达基因（DEGs）和多个富集信号通路，包括转化生长因子-β（TGF-β）受体信号激活 Smads、TGF-β 受体信号下调、TGF-β 受体复合物信号和胶原形成。此外，HXP 还显著抑制 ISO 诱导的原代大鼠 CFs 增殖、转分化、胶原合成以及 TGF-β1 和磷酸化 Smad2/3 蛋白表达的上调。
HXP 抑制 ISO 诱导的 CFs 转分化和胶原合成，其可能部分通过抑制 TGF-β/Smads 通路的激活来发挥作用。这可能是治疗心肌纤维化的一种新的治疗工具。
