Qin Hui, Wen Hao-Tian, Gu Kai-Juan, Hu Xu-Dong, Yang Tao, Yan Xiao-Feng, Ye Ting-Jie, Huo Jin-Lin, Hu Jing
Preclinical Medicine College, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People's Republic of China.
Seventh People's Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200137, People's Republic of China.
Drug Des Devel Ther. 2019 Aug 19;13:2873-2886. doi: 10.2147/DDDT.S185418. eCollection 2019.
Pulmonary fibrosis (PF) is a common clinical disease, which results in serious respiratory impairment. Xin Jia Xuan Bai Cheng Qi Decoction (XJXBCQ) is a traditional prescription commonly used in treating lung diseases. We investigate the effect of XJXBCQ against PF and its mechanism via the regulation of TGF-β1/Smad in vitro and in vivo.
XJXBCQ was first extracted and probed for chemical characterization. An PF model in vitro and in vivo was established in rats and in MRC-5 cells. In bleomycin (BLM)-induced rats model, lung function such as peak expiratory flow (PEF), minute ventilation (MV) and hydroxyproline (HYP) were measured; histopathological changes of lung tissue and TGF-β1 in peripheral blood of rats were detected. TGF-β receptor, Smad2 and its phosphorylation expression were tested by Western blot assay in rats model. Then the effects of XJXBCQ on TGF-β1/Smad signal pathway were assessed by Western blot analysis in vitro, and IL-17A and IL-25 levels were evaluated by ELISA in vivo.
Our results showed that XJXBCQ significantly enhanced the lung functions, such as PEF, MV and HYP, by reducing the expression level of lung inflammatory cytokine and the content and fibrosis of lung collagen. Moreover, XJXBCQ effectively inhibited TGF-β1, Smad2 and its phosphorylation expression, and the activation of Smad7 in vitro and in vivo. Furthermore, XJXBCQ had an inhibitory effect on the α-smooth muscle actin (α-SMA) and fibronectin (Fn) in vitro and downregulated IL-17A and IL-25 by inhibiting the activation of TGF-β1/Smad signaling pathway in vitro and in vivo. Further, XJXBCQ effectively inhibitied ventilation volume and peak expiratory content remodeling and hydroxyproline content through inhibition of TGF-βRⅡ, Smad2 and its phosphorylation expression, and activation of Smad7 in vivo.
XJXBCQ extract had an anti-PF effect in vitro and in vivo, which could be attributed to the inhibition of the expression of p-Smad2 and increase in the expression of Smad7 by regulating the TGF-β1/Smad activity.
肺纤维化（PF）是一种常见的临床疾病，可导致严重的呼吸功能损害。新加宣白承气汤（XJXBCQ）是治疗肺部疾病常用的传统方剂。我们通过在体外和体内调节转化生长因子-β1（TGF-β1）/Smad信号通路，研究XJXBCQ抗PF的作用及其机制。
首先提取XJXBCQ并进行化学表征分析。在大鼠和MRC-5细胞中建立体外和体内PF模型。在博来霉素（BLM）诱导的大鼠模型中，测量肺功能指标如呼气峰值流速（PEF）、分钟通气量（MV）和羟脯氨酸（HYP）；检测大鼠肺组织的组织病理学变化及外周血中TGF-β1水平。通过蛋白质免疫印迹法检测大鼠模型中TGF-β受体、Smad2及其磷酸化表达。然后在体外通过蛋白质免疫印迹分析评估XJXBCQ对TGF-β1/Smad信号通路的影响，在体内通过酶联免疫吸附测定法（ELISA）评估白细胞介素-17A（IL-17A）和白细胞介素-25（IL-25）水平。
我们的结果表明，XJXBCQ通过降低肺部炎性细胞因子的表达水平以及肺胶原蛋白的含量和纤维化程度，显著增强了肺功能，如PEF、MV和HYP。此外，XJXBCQ在体外和体内有效抑制了TGF-β1、Smad2及其磷酸化表达以及Smad7的激活。此外，XJXBCQ在体外对α-平滑肌肌动蛋白（α-SMA）和纤连蛋白（Fn）有抑制作用，并通过在体外和体内抑制TGF-β1/Smad信号通路的激活下调IL-17A和IL-25。此外，XJXBCQ在体内通过抑制TGF-βRⅡ、Smad2及其磷酸化表达以及Smad7的激活，有效抑制了通气量和呼气峰值含量重塑以及羟脯氨酸含量。
XJXBCQ提取物在体外和体内均具有抗PF作用，这可能归因于通过调节TGF-β1/Smad活性抑制p-Smad2的表达并增加Smad7的表达。
