Application for bioactivity-guided principle and prescription strategy to develop a new herbal formula in the treatment of pulmonary interstitial fibrosis

Autor: Tu,Yu-Ming, 涂祐銘
Druh dokumentu: 學位論文 ; thesis
Popis: 103
Pulmonary fibrosis contain chronic inflammatory disorder and complex progressive disease, which is associated with lung parenchyma injury and widespread cystic lesions including the alveolar wall, interstitial space, blood vessels, lymph and connective tissue around the trachea. Once a stimulus enters the lungs, initially cause lung epithelial cells or vascular endothelial damage, in which the tissue of the lung is destroyed so that trigger inflammation and cytokine dysregulation with massive release of proinflammatory cytokines. For pro-fibrotic effect, the expression of TGF-β is already induced during early stages of pulmonary fibrosis. TGF-β/Smad signaling is a major pathway leading to lung fibrosis, the striking ability of Smad proteins to accumulate in the nucleus in response to TGFβ was one of the early key observations placing Smads downstream of the TGFβ receptors. Smad proteins (eg. Smad3) undergo a constant process of nucleocytoplasmic shuttling, and their nuclear accumulation results from receptor-mediated phosphorylation events that enhance the myofibroblast differentiation and concurrent α-SMA overexpression (the excessive collagen production). It was well-known that bleomycin (BLM) easily causes cells to produce excess free radicals and eventually induce pulmonary fibrosis by apoptosis, has served as an early (inflammation) accompanied with a late (fibrosis) of acute lung injury in mice. In this study, a single intra-tracheal injection of BLM was conducted for inducing a fibrotic mouse model, resembling human pulmonary fibrosis. A critical evaluation of in vitro A549 (human lung adenocarcinoma epithelial cell line) model was utilized for developing a new herbal formula in the treatment of pulmonary interstitial fibrosis, following the bioactivity-guided principle and prescription strategy. Six traditional Chinese medicine, Polygonum cuspidatum, Ligusticum chuanxiong, Curcuma longa, Cordyceps sinensis,Astragalus membranaceus and Andrographis paniculata were extracted with 50% aqueous ethanol. For better anti-apoptotic bioactivities in A549 cell, four selected herbs will be formulated (named THAF-1) and administrated in pulmonary fibrosis of mice. After THAF-1 treatment, the physiological performance (body weight, survival) were observed, the biochemistry indicators (white blood cells, platelets, hs-CRP, HYP) was measured, pathological symptoms (inflammation, fibrosis) were examined, and the mechanisms underlying improved recovery of pulmonary fibrosis were elucidated by the expressions of TGF-β, Smad3, p-Smad3, COX-2, α-SMA. Our data showed that both 14-day THAF-1 (PO) and 5-days Vit.C (IP) can significantly inhibited the expression of TGF-β and the phosphorylation of Smad3, while THAF-1 (300mg/kg) had more reduction than Vit.C did on the expression of α-SMA by western blot. The results suggested that THAF-1 improve pulmonary fibrosis in mice might be partially via TGF-β/Smad3 signaling transduction pathways. In addition, THAF-1, as similar to the anti-inflammation effect of doxycycline/Vit.C, also profoundly decreased the COX-2 expression of lung tissue. Accordingly, THAF-1 (100 and 300mg/kg) could markedly ameliorate interstitial infiltration, increase alveolar ventilation and reduce the actual fibrosis by HE, Masson and IHC staining as compared to doxycycline. Taken together, the anti-fibrosis/anti-inflammation effects of THAF-1in BLM-induced fibrosis mice, suggesting that THAF-1 can be a potential candidate for botanical new drug development.
Databáze: Networked Digital Library of Theses & Dissertations
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