Inhibitory effect of Patrinia on BRL-3A cell apoptosis through the TLR4/PI3K/AKT/GSK3β and TLR4/P38/JNK signaling pathways.

The present study investigated the inhibitory effect of Patrinia on lipopolysaccharide (LPS)-induced apoptosis of rat liver BRL‑3A cells. A Cell Counting Kit‑8 assay was performed to measure the effect of Patrinia on BLR‑3A cell activities. A biochemical assay was employed to detect the release of lactate dehydrogenase (LDH) in BRL‑3A cells induced by different doses of LPS. Based on the release rate of LDH, drug concentrations were set at 0.5, 1 and 2 g/l. Apoptotic morphology of cells was observed via Hoechst 33342 staining and flow cytometry was performed to detect apoptosis rates. Western blotting was performed to detect the expression of toll‑like receptor 4 (TLR4), protein kinase B (AKT), phosphorylated (P)‑AKTSer473, glycogen synthase kinase 3β (GSK3β), P‑GSK3βSer9, P38, P‑P38, c‑Jun N‑terminal kinase (JNK), P‑JNK, B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2 associated X protein (Bax) and active‑caspase‑3 proteins. The translocation of GSK3β was observed by immunofluorescence staining. Results revealed that Patrinia increases cell activities and inhibits apoptosis. The expression levels of TLR4, P‑P38 and P‑JNK were reduced, whereas the expression of P‑AKTSer473 and P‑GSK3βSer9 were increased. Patrinia significantly reduced GSK3β nuclear translocation induced by LPS, and significantly decreased the mRNA expression levels of Bax/Bcl‑2 and caspase‑3 in BRL‑3A cells induced by LPS. In conclusion, Patrinia may significantly reduce apoptosis of BRL‑3A induced by LPS via the TLR4/PI3K/AKT/GSK3β and TLR4/P38/JNK signaling pathways, providing evidence for its potential use in liver disease therapy.