Dissection of the mechanism of traditional Chinese medical prescription-Yiqihuoxue formula as an effective anti-fibrotic treatment for systemic sclerosis

Abstract

BackgroundSystemic sclerosis (SSc) is a connective tissue fibrotic disease for which there is no effective treatment. Traditional Chinese Medicine (TCM), such as the Yiqihuoxue formula used in Shanghai TCM-integrated Hospital, has shown the efficacy of anti-fibrosis in clinical applications. This study was aiming to dissect the anti-fibrotic mechanism of Yiqihuoxue treatment for SSc.MethodsBleomycin-induced mice and SSc dermal fibroblasts were treated with Yiqihuoxue decoction; NIH-3T3 fibroblasts were exposed to exogenous TGF-β1, and then cultured with or without Yiqihuoxue decoction. Luciferase reporter gene assay was used to determine the activity of Smad binding element (SBE). Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the mRNA levels of extracellular matrix (ECM) genes. The protein levels of type I collagen, Smad3 and phosphorylated-Smad3 (p-Smad3) were detected by western blotting. Student’s t-tests were used to determine the significance of the results.ResultsBleomycin-induced mice, SSc dermal fibroblasts and TGF-β1-induced NIH/3T3 fibroblasts showed higher levels of ECM gene transcriptions and collagen production. In addition, the phosphorylation level of Smad3 and activity of SBE were significantly increased after exogenous TGF-β1 induction. Whereas, Yiqihuoxue treatment could obviously attenuate fibrosis in bleomycin-induced mice, down regulate ECM gene expressions and collagen production in SSc dermal fibroblasts and TGF-β1-induced NIH/3T3 fibroblasts. Furthermore, the aberrantly high phosphorylation level of Smad3 and activity of SBE in the TGF-β1-induced NIH/3T3 fibroblasts were also dramatically decreased by Yiqihuoxue treatment.ConclusionsYiqihuoxue treatment could effectively reduce collagen production via down-regulating the phosphorylation of Smad3 and then the activity of SBE, which are involved in the TGF-β pathway and constitutively activated in the progression of SSc.

Increased expression of latent TGF-β-binding protein 4 affects the fibrotic process in scleroderma by TGF-β/SMAD signaling

“…The animals used in experiment were approved by the School of Life Sciences, Fudan University, China, and the animal protocols were performed in accordance with the approved guidelines. 21…”

Salvianolic Acid B Attenuates Experimental Pulmonary Fibrosis through Inhibition of the TGF-β Signaling Pathway

“…The Yiqihuoxue formula was prepared as previously described15. pGL3-SBE 4 -Luc was a gift from Dr. Kiyoshi Higashi (Sumitomo Chemical Co., Ltd., Osaka, Japan), and consists of four short tandem repeats of the SBE oligonucleotide (GTCTAGAC) inserted into the pGL3 Basic vector35.…”

Traditional Japanese formulas tokishakuyakusan and ogikenchuto suppress dermal sclerosis in bleomycin‐induced murine scleroderma

“…The other parts were immediately stored in liquid nitrogen. To evaluate skin sclerosis, a 4 μm section of skin was stained using hematoxylin and eosin; its thickness was then measured, using Image J (National Institutes of Health, Bethesda, MD, USA), on photographs taken in 10 microscopy fields (×100) at the site of maximum distance between the epidermal-dermal and dermal-subcutaneous fat junctions [11]. The mean AE SD of these measurements was then calculated.…”

“…In particular, dose-dependent KBG treatment has been shown in vitro to inhibit collagen production from cultured fibroblasts of SSc patients' skin [10]. Recently, it was reported that yiqihuoxuetang, which contains astragali radix, suppressed collagen production and dermal sclerosis progression in a bleomycin (BLM)-induced mouse model of scleroderma [11].…”