Artesunate modulates expression of matrix metalloproteinases and their inhibitors as well as collagen-IV to attenuate pulmonary fibrosis in rats

Abstract

ABSTRACT. The aim of this study was to determine the effect of artesunate on extracellular matrix (ECM) accumulation and the expression of collagen-IV, matrix metalloproteinase (MMP), and tissue inhibitor of matrix metalloproteinase (TIMP) to understand the pharmacological role of artesunate in pulmonary fibrosis. Eighty Sprague-Dawley rats were randomly assigned to four groups that were administered saline alone, bleomycin (BLM) alone, BLM + artesunate, or artesunate alone for 28 days. Lung tissues from 10 rats in each group were used to obtain lung fibroblast (LF) primary cells, and the rest were used to analyze protein expression. The mRNA expression of collagen-IV, MMP-2, MMP-9, TIMP-1, and TIMP-2 in lung fibroblasts was detected by real-time quantitative reverse transcriptase polymerase chain reaction. The protein levels of collagen-IV, MMP-2, MMP-9, TIMP-1, and TIMP-2 protein in lung tissues were analyzed by western blotting. Artesunate treatment alleviated alveolitis and pulmonary fibrosis induced by bleomycin in rats, as indicated by a decreased lung coefficient and improvement of lung tissue morphology. Artesunate treatment also led to decreased collagen-IV protein levels, which might be a result of its downregulated expression and increased MMP-2 and MMP-9 protein and mRNA levels. Increased TIMP-1 and TIMP-2 protein and mRNA levels were detected after artesunate treatment in lung tissues and primary lung fibroblast cells and may contribute to enhanced activity of MMP-2 and -9. These findings suggested that artesunate attenuates alveolitis and pulmonary fibrosis by regulating expression of collagen-IV, TIMP-1 and 2, as well as MMP-2 and -9, to reduce ECM accumulation.

Artesunate ameliorates lung fibrosis via inhibiting the Notch signaling pathway

“…Lung fibroblasts were isolated from the rats described below according to a previous method ( 13 ). The cells were cultured (at 37°C, 95% humidity and 5% CO 2 ) in DMEM supplemented with 10% FBS.…”

“…Cultured cells were lysed on ice in ice-cold RIPA buffer with a protease inhibitor cocktail containing 1 mmol/l PMSF for 20 min. Proteins from lung tissue and fibroblast were prepared as previously described ( 12 , 13 ). A total of 20 µg protein lane was loaded into each lane and separated via 10% SDS-PAGE gel and then transferred to a polyvinylidene difluoride membrane, which was blocked with 5% non-fat milk in Tris-buffered saline (TBS) at room temperature for 1.5 h. The membranes were then incubated at 4°C overnight with cleaved Notch (NICD 1:1,000 dilution; cat.…”

Early Plasma Matrix Metalloproteinase Profiles. A Novel Pathway in Pediatric Acute Respiratory Distress Syndrome

“…Interestingly, both animals and humans with bleomycininduced pulmonary fibrosis have demonstrated decreased MMP-9/TIMP-1 ratios, and treatment with the antifibrotic drug pirfenidone has been associated with restoration of the MMP-9/TIMP-1 ratio in the lungs, downregulation of the TGF-b pathway, and clinical improvement (18,(58)(59)(60)(61). Other pharmacotherapeutics that have reported efficacy in modulating the MMP-9/TIMP-1 ratio in patients with pulmonary fibrosis include aliskiren, artusenate, bosentan, doxycycline, and induced pluripotent stem cells, although further studies are needed (62)(63)(64)(65)(66).…”

Resveratrol raises in vitro anticancer effects of paclitaxel in NSCLC cell line A549 through COX-2 expression

“…TIMP-3 could inhibit the activities of MMP-1, MMP-2, MMP-3 and MMP-9, high TIMP-3 level could inhibit the cancer [ 24 ]. TIMP-1/2 inhibits the degradation of ECM by inhibiting the activation of MMP-2/9, which plays an important role in the formation of pulmonary fibrosis, pulmonary fibrosis may develop into lung cancer [ 25 ].…”