LncRNA MEG3 suppresses PI3K/AKT/mTOR signalling pathway to enhance autophagy and inhibit inflammation in TNF-α-treated keratinocytes and psoriatic mice

Background: Psoriasis is a common chronic inflammatory skin disorder that causes patches of thick red skin and silvery scales and affects 1-3% of the population, which reduces patient's quality of life. Understanding the pathogenesis of psoriasis is crucial for developing novel therapeutic strategies. Methods: HaCaT and NHEK cells were treated with TNF-alpha in vitro. A mouse model of psoriasis was established by topical imiquimod application on back skin. LncRNA MEG3 was cloned into the pcDNA3.1 vector and transfected in TNF-alpha-treated HaCaT and NHEK cells to overexpress its expression. Liposome-encapsulated pcDNA3.1-MEG3 was injected into imiquimod-treated mice via tail vein. RT-qPCR and western blot assays were used to examine the expression of lncRNA MEG3, IL-6, IL-8, IFN-gamma, IL-1 beta, LC3, Beclin 1, p62, p-p65, p65, NLRP3, p-PI3K, PI3K, pAKT, AKT, p-mTOR, mTOR respectively. The secretion of IL-6, IL-8, IFN-gamma and IL-1 beta was determined using ELISA assay. Immunofluorescence and immunohistochemistry methods were performed for analyzing the expression of LC3 and NLRP3 in cells and skin tissues respectively. LY294002 was used to block the PI3K/AKT/mTOR signalling. MTT assay was applied to test the toxicity of LY294002 to HaCaT and NHEK cells. Results: LncRNA MEG3 expression levels were downregulated in TNF-alpha-treated HaCaT and NHEK cells and skin tissues of psoriatic mice model. TNF-alpha treatment enhanced inflammation and suppressed autophagy in HaCaT and NHEK cells, which were largely reversed by overexpression of lncRNA MEG3. Autophagy puncta and NLRP3 inflammasome assembly showed the same patterns with the expression of inflammation and autophagy markers in TNF-alpha-treated HaCaT and NHEK cells with or without lncRNA MEG3 overexpression. TNF-alpha-induced activation of the PI3K/AKT/mTOR signalling was abolished by lncRNA MEG3 overexpression in HaCaT and NHEK cells. Blocking the PI3K/AKT/mTOR signalling inhibited TNF-alpha-induced inflammation and restored autophagy level in TNF-alpha-treated HaCaT and NHEK cells. Overexpression of lncRNA MEG3 suppressed inflammation, promoted autophagy and inhibited the activation of the PI3K/AKT/mTOR signalling in a mouse model of psoriasis. Conclusion: LncRNA MEG3 facilitates autophagy and suppresses inflammation in TNF-alpha-treated keratinocytes and psoriatic mice, which is dependent on the PI3K/AKT/mTOR signalling pathway. Our study enhances the understanding of psoriasis and provides potential therapeutic targets for psoriasis.

Automated summary

The study reveals that downregulation of lncRNA MEG3 is associated with enhanced inflammation and suppressed autophagy in psoriasis, while overexpression of lncRNA MEG3 can reverse these effects, potentially through the PI3K/AKT/mTOR signaling pathway. Therefore, lncRNA MEG3 may serve as a therapeutic target for psoriasis.