Puerarin attenuated ischemia/reperfusion injury an in vitro model through a repression of autophagy by HMGB1/TLR4/NF-κB axis / 中华器官移植杂志

ABSTRACT

Objective:To explore the protective effect of puerarin on hypoxia/reoxygenation (H/R)-induced acute kidney injury(AKI)in vitro.Methods:HK-2 cells were treated with H/R for simulating ischemia reperfusion injury(IRI)in vivo. The experimental groups included control group, H/R treatment group(0/6/12/24 h), H/R 24 h + puerarin treatment group(puerarin, Pue), H/R 24 h + Pue+ 3-methyladenine(3-MA)treatment group and H/R 24 h+ 3-MA treatment group. Immunoblotting was employed for detecting the expression changes of autophagy-related proteins, CCK-8 for examining cell proliferation, electron microscopy for observing autophagosome formation and TUNEL for detecting apoptosis.Results:As compared with control group, the expression of LC3-II rose in H/R 24 h group, the expression of autophagy marker P62 declined, count of autophagosome increased, cell viability decreased and cellular inflammation occurred. Puerarin had similar effects to 3-MA. As compared with H/R 24 h group, puerarin could reverse the changes in the expression levels of LC3 and P62 induced by H/R( P<0.05). There were greater cell viability, reduced autophagosome count and lessened cell apoptosis( P<0.05). At the same time, protein expression levels of HMGB1, TLR4 and NF-κB dropped( P<0.05). Conclusions:Puerarin suppresses autophagy through HMGB1/TLR4/NF-κB axis for lessening ischemia-reperfusion injury an in vitro model.