Mechanism of 6% Hydroxyethyl Starch HES130/0.4 Regulating PGC-1α Expression and Inhibiting LPS-Induced Apoptosis of RAW264.7 Cells

Abstract

This study mainly observed the inhibitory effect of 6% hydroxyethyl starch 130/0.4 (HES) on the apoptosis of RAW264.7 cells induced by lipopolysaccharide (LPS) and its effect on the peroxisome proliferator-activated receptor coactivator-1α. Screening the concentration of HES by CCK-8; Determination of target genes of HES by gene chip technology. Cell experiments were divided into Control group, LPS group, HES group and PGC-1α-shRNA group. The growth activity of cells in each group was detected by CCK-8 method and the apoptosis rate of cells in each group was detected by flow cytometry. Rh123 staining was used to detect the membrane potential of mitochondria in each group Western blot was used to detect PGC-1α, nuclear respiratory factor 1 (NRF1) and NRF2, OpticAtrophy1 (OPA1) and mitochondrial outer membrane fusion protein 2 (mitofusin2, MFN2) and the expression of B cell lymphoma-2 (Bcl-2) and Bcl-2 related X protein (Bax). The concentration of HES was (30 mg/mL). It was verified by gene chip and PCR that the target of HES was PGC-1α; Compared with the cells in Control group, the growth activity, mitochondrial membrane potential, expressions of PGC-1α, NRF2, NRF1, OPA1, MFN2 and Bcl-2 in LPS group, HES group and PGC-1α-shRNA group decreased significantly, while the apoptosis rate and the expression of Bax in cells increased significantly. Compared with the cells in LPS group, the growth activity, mitochondrial membrane potential, the expressions of PGC-1α, NRF2, NRF1, OPA1, MFN2 and Bcl-2 in HES group and PGC-1α-shRNA group increased significantly, while the apoptosis rate and the expression of Bax in cells decreased significantly. Compared with HES cells, the growth activity of PGC-1α-shRNA cells, the membrane potential of mitochondria, the expressions of PGC-1α, NRF2, NRF1, OPA1, MFN2 and Bcl-2 in cells decreased significantly, and the apoptosis rate and the expression of Bax in cells increased significantly, with statistical significance (all P<0.05). 6% hydroxyethyl starch 130/0.4 (HES) could regulate the expression of PGC-1α, enhance the biosynthesis of mitochondria in cells, and inhibit the apoptosis of RAW264.7 cells induced by LPS.