Protective Effect Of 8-formylophiopogonanone B On Cardiotoxicity Induced By Doxorubicin

Background:Doxorubicin(Dox)is an anthracycline chemotherapy drug widely used in clinical applications,but it is restricted due to cardiotoxicity.8-Formylophiopogonanone B(8-FOB)is a flavonoid drug isolated from traditional Chinese medicine Ophiopogon japonicushas cardiovascular protective effects.However,whether 8-FOB can reduce Dox-induced cardiotoxicity and its possible mechanism is not clear.Objective:The purpose of this study is to explore whether 8-FOB could protect against the acute cardiotoxicity induced by Dox and the possible mechanism.Methods:To establish an acute cardiotoxicity,mice received a single intraperitoneal injection of Dox(20 mg/kg).We screened the hub genes associated with Dox-induced acute cardiotoxicity via bioinformatics analysis.And then,we verified the expression of hub gene with western blot and real-time quantitative PCR.In addition,mice were randomly divided into four groups(15 mice per group):Control group,Dox group,8-FOB group and 8-FOB+Dox group.Control group:mice in the control group received the same volume of normal saline or 0.5%CMC-Na suspension as the experimental group;Dox group:mice were given a single intraperitoneal injection of Dox(20 mg/kg);8-FOB group:mice were gavaged 8-FOB suspension(20 mg/kg/day);8-FOB+Dox group:mice were given a single intraperitoneal injection of Dox(20 mg/kg)and gavaged 8-FOB(20 mg/kg/day).Among them,8-FOB was administered for five consecutive days.Five days after Dox treatment,histological analysis(hematoxylin-eosin(H&E)staining,Masson staining),biochemical assays(creatine kinase-MB,CK-MB),(Lacticde hydrogenase,LDH),(Hydroxybutyrate Dehydrogenase,HBDH),cardiac function(left ventricular ejection fraction percentage,EF%),(fractional shortening percentage,FS%),(Heart Rate,HR),pro-inflammatory cytokines(Interleukin-1β,IL-1β),(tumor necrosis factor-α,TNF-α),(Interleukin-6,IL-6)were conducted to evaluate the protective effect of 8-FOB on acute cardiotoxicity induced by Dox.Results:Compared with control group,Dox induced cardiac dysfunction(decreased EF%,FS%and HR(P<0.01);serum CK-MB(P<0.01),LDH and HBDH levels were increased after Dox administration(P<0.05),H&E staining showed disordered myofibrillar after Dox administration;Masson staining showed that Dox increased the level of myocardial fibrosis,IL-1β(P<0.05)and IL-6(P<0.01)mRNA levels were also increased.Bioinformatics analysis and real-time quantitative PCR and western blot results showed that hub gene Hmxo 1 was up-regulated after Dox administration(P<0.05).In addition,Hmox1 was involved in the positive regulation of I-κB kinase/NF-κB signaling pathway.Compared with Dox group,8-FOB reversed the cardiac dysfunction,pathological changes and the increases in serum CK-MB,LDH and HBDH caused by Dox(P<0.01).Compared with control group,the collagen content in heart tissue was increased,and IL-1β and IL-6 mRNA were also increased(P<0.05)after Dox administration.Compared with Dox group,8-FOB significantly reduced the cardiac collagen content and IL-β(P<0.01)and IL-6 mRNA(P<0.05).In addition,western blot results showed that Dox induced a significant increase in the expression of HMOX1(P<0.01),while compared with the Dox group,the HMOX1 expression in the Dox+8-FOB group was significantly reduced(P<0.01).Conclusions:Hmxo1 as a hub gene is up-regulated and positive regulates the inflammatory signaling pathway in Dox-induced cardiotoxicity.In addition,8-FOB protects against Dox-induced acute cardiotoxicity by ameliorating Dox-induced cardiac fibrosis and inflammation,and inhibiting Hmoxl expression.The cardioprotective effect of 8-FOB may depend on inhibiting Hmoxl expression.The results of this study lay a foundation for further exploration of the mechanisms underlying Dox-induced cardiotoxicity and protective agents that can combat it.