Panoply™ Human PDPK1 Over-expressing Stable Cell Line

The health risks associated with alcohol consumption are very serious, sometimes even life-threatening. Alcoholic steatohepatitis (ASH) is a severe stage leading to cirrhosis and end-stage liver disease. However, its pathogenesis is still not fully understood, and there is currently no universally accepted effective treatment. Here, researchers observed a significantly increased expression level of 3-phosphoinositide-dependent protein kinase 1 (PDPK1, also known as PDK1) in an ASH model. Therefore, PDPK1 may influence the progression of ASH. The researchers further examined the autophagy flux under PDPK1 changes to investigate whether PDPK1 can regulate cellular autophagy in ASH. The study showed that PDPK1 affects hepatocyte self-repair by inhibiting autophagy. Inhibiting both PDPK1 and its phosphorylation (at the ser241 site) protected hepatocytes from severe alcoholic hepatitis damage.

Here, researchers constructed PDPK1-overexpressing stable cell lines. Transfection efficiency was detected by RT-qPCR and Western blotting (Figure 1A, B). Both Western blotting and confocal microscopy observations showed that autophagy flux was inhibited in PDPK1-overexpressing cells (Figure 1C, D). Consistent with this, Nile Red staining confirmed that PDPK1 overexpression further increased ethanol-induced lipid vacuole accumulation (Figure 1E).

Figure 1. Upregulated PDPK1 expression promoted steatosis and inhibits autophagy in hepatocytes. (Zhang Y, et al., 2022)