MIRO1-modified MSC hUCMSCs in repairing lung epithelial cell injury through mitochondrial autophagy

Background Mesenchymal stem cells (MSCs) provide a new strategy for the treatment and repair of acute lung injury (ALI). Currently, MSCs alone have poor therapeutic effect, and methods to enhance the efficacy of MSCs need to be explored.

Objective To investigate the therapeutic effect of mitochondrial Rho GTPase 1(Rhot1) overexpression of human umbilical cord mesenchymal stem cells (hUCMSCs) on lipopolysaccharide-induced injury of human lung bronchial epithelial cells (BEAS-2B).

Methods Lentivirus overexpressing MIRO1 was packaged and hUCMSCs overexpressing MIRO1 were constructed by lentivirus infection. The experimental cells were randomly divided into control group, lipopolysaccharide group (LPS), lipopolysaccharide + human umbilical cord mesenchymal stem cells (Con-hUCMSCs) group and lipopolysaccharide + human umbilical cord mesenchymal stem cells overexpressing MRIO1 (MIRO1-hUCMSCs) group. Lipopolysaccharide stimulated BEAS-2B cells for 24h and co-cultured with Con-hUCMSCs and MIRO1-hUCMSCs cells through Transwell chamber for 24h. The BEAS-2B cells were collected, and tumor necrosis factor-α (TNF-α) and interleukin (1β) were detected by Western blot and RT-qPCR. The expression levels of IL-1β, interleukin (IL-6), and mitochondrial autophagy associated proteins (PINK1, PARKIN, MIRO1, SQSTM1) were leukin.

Results hUCMSCs stable cell lines overexpressing MIRO1 were successfully screened. Through Transwell cell co-culture, MIRO1-hUCMSCs group inhibited the increase of inflammatory factors TNF-α, IL-1β and IL-6 in LPS group (P<0.05), restored the low expression of mitochondrial autophagy proteins PINK1, SQSTM1 and MIRO1 (P<0.05), decreased the high expression of PARKIN protein (P<0.05). The effect in MIRO1-hUCMSCs group showed statistical significance compared with LPS group and Con-hUCMSCs group (P<0.05).

Conclusion Compared with hUCMSCs alone, MiRO1-modified hUCMSCs can ameliorate lipopolysaccharids-induced BEAS-2B cell damage by regulating PINK1, PARKIN, MIRO1 and SQSTM1 proteins in mitochondrial autophagy Pink1-Parkin pathway.