Autophagy Negatively Regulates Osteogenic Phenotype Transformation of Porcine Aortic Valvular Interstitial Cells Induced by LPS
ZHANG Menghao, FAN Mengtian, AN Liqin, CHEN Bin, WU Jinghong, WANG Yue, HUANG Qin, SHI Qiong*
This article mainly investigated the effect and mechanism of autophagy on osteogenic phenotype transformation of valvular interstitial cells induced by LPS (lipopolysaccharide). The expression level of osteogenesis, inflammation and autophagy in calcific (3 cases) and normal (3 cases) aortic valve samples were detected by immunohistochemistry. LPS was used to treat pAVICs (porcine aortic valvular interstitial cells) (2-6 generations), and the protein levels of osteogenesis, inflammation and autophagy were detected using Western blot. Autophagy agonists and antagonists were combined with LPS to treat the primary pAVICs, and the changes of autophagy, osteogenesis and inflammation were detected by Western blot. Autophagy activation was detected by transfecting GFP-LC3. NF-κB antagonists combined with LPS to treat pAVICs, and the changes of inflammation and osteogenesis were detected by Western blot. Autophagy agonists and antagonists were combined with LPS to treat the primary pAVICs, and levels of inflammation were detected by Western blot. The results showed that the expression of the osteogenic index (RUNX2, OPN), inflammatory index (p-NF-κB) and autophagy index (LC3) in the lesion aortic valve specimens were higher than those in the normal control group. Western blot results showed an up-regulation of osteogenesis, inflammation and autophagy in primary pAVICs treated by LPS. Autophagy antagonists can inhibit autophagy activation and promote the up-regulation of osteogenesis induced by LPS. Autophagy agonists can promote autophagy activation and significantly inhibit the up-regulation of osteogenesis induced by LPS. NF-κB antagonists can inhibit inflammation and osteogenesis induced by LPS. Autophagy agonists and antagonists can inhibit and promote the up-regulation of inflammation induced by LPS, respectively. It is possible that autophagy negatively regulates the osteogenic phenotype transformation of VICs induced by LPS, which may play a regulatory role by inhibiting the inflammatory pathway, NF-κB.
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