Study On PK-PD Model Of Anti-inflammatory Effects Of Rutin In Mice With Acute Lung Injury Induced By Lipopolysaccharide

As the breeding industry is developing rapidly in recent years,the incidence of inflammatory diseases is increasing as well.For instance,infectious diseases,parasites,medical and surgical diseases can trigger inflammatory lesions.Acute inflammation is available to affect the health of livestock and poultry or even endanger their life,further affect livestock production and cause economic losses.Rutin is a kind of flavonoid with many pharmacological activities such as anti-oxidation,anti-inflammation,antiviral,etc.However,there are great limitations in veterinary clinical application because of its poor solubility and slow absorption in vivo.In this study,a pharmacokinetics and pharmacodynomics（PK-PD）model was adopted to study the correlation between the pharmacokinetic characteristics of rutin in vivo and the pharmacodynomic on inflammation in acute lung injury.In this study,a mouse model of lipopolysaccharide（LPS）-induced acute lung injury was applied and rutin was intragastrically administered continuously for 5 days（200 mg/kg/d）followed by the mice sacrificed on days 2,3,4,5,and 6 of drug administration to dissect the lung tissues.One of which was used to calculate the lung wet-to-dry weight ratio,and the other was used to determine the NO content and protein expression level;the NO content and the changes in TLR4,TRAF6,IκB,P-IκB protein expression levels in the inflammatory signaling pathway were detected by Griess method and Western blotting method,respectively.The results showed that rutin was available to significantly inhibit the synthesis of inflammatory mediators（NO）through directly anti-inflammatory and protective effect,moreover,it could inhibit the expression of TLR4,TRAF6 protein levels and the phosphorylation of IκB and p65 through anti-inflammatory effect.In this study,luteolin was applied as an internal standard to establish a high-performance liquid chromatographic method for the determination of the desired concentration of rutin in plasma.After the acute lung injury model was established in mice,rutin was intragastrically administered for 5 consecutive days（200 mg/kg/d）.In addition,blood samples were collected from the posterior ocular plexus vein on days 1,2,3,4 and 5 of administration at 3,6,12,18,21,24,27,30,36,48,60,and 72 h.The plasma concentrations of rutin were measured by the above established high performance liquid chromatography method,and the pharmacokinetic model of rutin was fitted using Win Nonlin software with intention to fit the concentration-time curve of rutin and calculate the pharmacokinetic parameters.The absorption of rutin in mice with lipopolysaccharide-induced acute lung injury conformed to a one-compartment model,with T_1/2α=9.755 h,T_1/2β=19.442 h,T_max=24.00 h,C_max=22.651μg/m L,AUC=518.577μg/m L·h。A PK-PD combination model was established to fit the optimal administration time of rutin with a one-compartment-Sigmod Emax model connected to the effect site;meanwhile,the effects of rutin on NO content and protein level expression of TLR4,TRAF6,IκB,P-IκB were revealed by the combination model,and the relationship between the changes of the above indicators and the kinetic characteristics of rutin was described.In this study,lipopolysaccharide-induced acute lung injury was applied as a model to study the kinetics and pharmacodynamics of rutin in mice with acute lung injury,and a PK-PD combination model of rutin in the treatment of acute lung injury was established on this basis.The correlation between the pharmacokinetics and pharmacodynamics of rutin in mice with lipopolysaccharide-induced acute lung injury was quantitatively analyzed to provide a theoretical basis for the research and development of new anti-inflammatory drugs in veterinary clinical practice.