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Effect of AMPK mutants on BMP9-induced proliferation and differentiation of HFL-1 cells. (A) CCK-8 assay was used to detect the effect of AMPK-CA on the proliferation of HFL-1 cells induced by BMP9 (n = 3). (B) CCK-8 assay was used to examine the role of AMPK-DN in the inhibitory effect of metformin on BMP9-induced proliferation of HFL-1 cells (n = 3). (C,D): The roles of AMPK-CA in the protein expression of fibroblast differentiation markers (ɑ-SMA, Collagen I and Collagen III) were detected by western blot (n = 3). (E,F): The effects of AMPK-DN on the protein expression of ɑ-SMA, Collagen I and Collagen III were tested by western blot (n = 3). Data are presented as mean ± SD. **p < 0.01 versus only GFP-AD group, # p < 0.05 and ## p < 0.01 versus GFP-AD with BMP9 group, && p < 0.01 versus GFP-AD with BMP9 and metformin group.
Source publication
Adenosine monophosphosphate-activated protein kinase (AMPK) and its activator metformin were found to be involved in the regulation of fibroblast activation and pulmonary fibrosis. However, the regulatory mechanism has been undetermined. Recently, AMPK has been reported to exert its effect through inhibiting bone morphogenetic protein (BMP) pathway...
Contexts in source publication
Context 1
... one thing, we examined the effect of AMPK-CA on the proliferation of HFL-1 cells by CCK-8 assay. As shown in Figure 3A, the proliferative abilities of Western blot was applied to test the expression of p-AMPK, ALK1 and p-Smad1/5 in HFL-1 cells with 10 mM metformin for different times (2, 4, 8 and 24 h) and 10 ng/ml BMP9 for 0.5 h. Cells dealt with culture medium were served as control group (n = 3). ...
Context 2
... of metformin on BMP9-induced proliferation of HFL-1 cells. Results showed that AMPK-DN reversed the effect of metformin on BMP9-induced HFL-1 cells proliferation, as compared to GFP-AD with BMP9 group ( Figure 3B). Meanwhile, we tested the effect of AMPK-CA and AMPK-DN on the protein expressions of fibroblast differentiation markers, including ɑ-SMA, Collagen I and Collagen III, using western blot. ...
Context 3
... we tested the effect of AMPK-CA and AMPK-DN on the protein expressions of fibroblast differentiation markers, including ɑ-SMA, Collagen I and Collagen III, using western blot. Western blot assay indicated that AMPK-CA restrained the up-regulation of these proteins induced by BMP9 in HFL-1 cells ( Figures 3C,D), while AMPK-DN counteracted the response of metformin to the expressions of ɑ-SMA, Collagen I, and Collagen III in BMP9-treated HFL-1 cells (Figures 3E,F). These results suggest that metformin activates AMPK, Western blot was applied to detect the effect of AMPK-DN on the expression of ALK1 and p-Smad1/5 (n = 3). ...
Context 4
... we tested the effect of AMPK-CA and AMPK-DN on the protein expressions of fibroblast differentiation markers, including ɑ-SMA, Collagen I and Collagen III, using western blot. Western blot assay indicated that AMPK-CA restrained the up-regulation of these proteins induced by BMP9 in HFL-1 cells ( Figures 3C,D), while AMPK-DN counteracted the response of metformin to the expressions of ɑ-SMA, Collagen I, and Collagen III in BMP9-treated HFL-1 cells (Figures 3E,F). These results suggest that metformin activates AMPK, Western blot was applied to detect the effect of AMPK-DN on the expression of ALK1 and p-Smad1/5 (n = 3). ...
Citations
... Thalidomide and other IMiDs such as lenalidomide and pomalidomide inhibit the action of cereblon and allow the expression of AMPK . Inhibition of AMPK is one of the mechanisms by which metformin inhibits proliferation and differentiation of human foetal lung fibroblast (Gu et al. 2021;Chen et al. 2022). AMPK modulation through cereblon shows a protective action and slows down the growth of fibrogenic proteins such as fibronectin, collagen α-SMA. ...
The “Thalidomide tragedy” is a landmark in the history of the pharmaceutical industry. Despite limited clinical trials, there is a continuous effort to investigate thalidomide as a drug for cancer and inflammatory diseases such as rheumatoid arthritis, lepromatous leprosy, and COVID-19. This review focuses on the possibilities of targeting inflammation by repurposing thalidomide for the treatment of idiopathic pulmonary fibrosis (IPF). Articles were searched from the Scopus database, sorted, and selected articles were reviewed. The content includes the proven mechanisms of action of thalidomide relevant to IPF. Inflammation, oxidative stress, and epigenetic mechanisms are major pathogenic factors in IPF. Transforming growth factor-β (TGF-β) is the major biomarker of IPF. Thalidomide is an effective anti-inflammatory drug in inhibiting TGF-β, interleukins (IL-6 and IL-1β), and tumour necrosis factor-α (TNF-α). Thalidomide binds cereblon, a process that is involved in the proposed mechanism in specific cancers such as breast cancer, colon cancer, multiple myeloma, and lung cancer. Cereblon is involved in activating AMP-activated protein kinase (AMPK)-TGF-β/Smad signalling, thereby attenuating fibrosis. The past few years have witnessed an improvement in the identification of biomarkers and diagnostic technologies in respiratory diseases, partly because of the COVID-19 pandemic. Hence, investment in clinical trials with a systematic plan can help repurpose thalidomide for pulmonary fibrosis.
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