Puerarin Attenuates NOX-dependent Oxidative And Inflammatory Responses Induced By Acute Cigarette Smoke Exposur

Puerarin,an active ingredient of the Chinese traditional medicine,has been shown to exert anti-oxidant and anti-inflammatory effects in the cardiovascular system.The effects of Puerarin on early oxidative and inflammatory responses in ACS-induced mice and on CSE-treated HSAECs were investigated in the current study.The C57/6J mice were ACS exposed for 1 hour and lung tissues were harvested at 2,6 and 24 hours after ACS exposed,with or without Puerarin(40 or 80 mg/kg)treatment.ACS-induced superoxide production significant increased at 2,6 hours after ACS exposed and dose-dependently abrogated by Puerarin in mice lung sections by DHE fluorescent imaging,and Puerarin had no effect on buffering superoxide in a cell free system determined by ERS.Protein expressions of NOX1,2 and 4 were up-regulated in ACS exposed mice which were abolished by Puerarin.Further,neutrophils and macrophages were significantly increased in both lung parenchyma and BAL fluid of ACS exposed mice,and the recruitment of inflammatory cells in lung parenchyma and BAL fluid were suppressed by Puerarin.The upregulations of mRNA expressions(TNF-α and KC)and protein expressions(phospho-p65,TNF-α and COX-2)in ACS-exposed mice were partly abrogated by Puerarin-treated mice in a dose-dependent manner.In vitro,superoxide or hydrogen peroxide productions increased in CSE stimulated HSAECs in time and concentration dependent manners which were tested by DHE and DCFH-DA fluorescent imaging.The protein expressions of NOX1,2,4 and 5 were up-regulated in CSE-induced HSAECs.Importantly,Puerarin significant reversed ROS productions and the protein expressions of NOX1 and 4 in CSE treated HSAECs.Further,RelA/p65 nuclear translocations increased in CSE-induced HSAECs which were attenuated by Puerarin which detected by immunofluorescence microscopy analyses.CSE increased IL-8 secretions in culture medium and upregulated the protein expression of COX-2 in HSAECs.Likewise,the IL-8 secretions and the protein expression of COX-2 were dose-dependently reversed by Puerarin in CSE induced HSAECs.Moreover,ACS-induced ROS productions and the expression of COX-2 were partially rescued by ROS scavenger(TEMPO)in vivo and vitro,with no additional synergy for combinatory treatment with Puerarin and TEMPO.Collectively,Puerarin attenuated upregulation of NOX1,2 and 4 in ACS-exposed mice,and upregulation of NOX1 and 4 in CSE-treated HSAECs,leading to inhibition of ROS production and the activation of ROS-sensitive NF-κB signaling pathway which attenuate the protein expression of COX-2.These data support the novel therapeutic value of Puerarin in cigarette smoke induced oxidative and inflammatory responses in early COPD.