| Objective: Acute lung injury is a typical inflammatory disease,which is characterized by an intense pulmonary inflammatory response,including the accumulation of neutrophils,the release of inflammatory mediators,the increase of pulmonary vascular permeability and pulmonary interstitial edema.The pathogenesis of ALI can be summarized as the damage of immune system and over activation of complement system.ALI,in more serious instances,can lead to acute respiratory distress syndrome,remains a severe disease and still presents the high mortality rate of approximately 40%.Despite the ever deepening understanding of ALI/ARDS pathophysiology,effective therapies to control the morbidity and mortality of the disease are scarce.Therefore,the development of new mediation and new strategies for the management of this disease is urgently needed.Matrine is one of the main active components of Chinese herb Sophora flavescens Ait(Kushen),which has been demonstrated to be effective in suppressing inflammation.Meanwhile,lycopene is one of the most important components of carotene.It has many important physiological functions,such as antioxidation,scavenging free radicals and so on.In order to explore the effective treatment of acute lung injury and provide reference for application in clinic,we investigated the protective and therapeutic effects of matrine and lycopene combined on lipopolysaccharide-stimulated acute lung injury in mice and its mechanism.Methods: 120 Kun Ming mouse were divided into 6 groups(n=20)randomly: normal control(NM)group,model(LPS)group,dexamethasone(DEX)group,matrine group,lycopene group,matrine and lycopene(UNION)group.NM group and LPS group’s mouse were pretreated with normal saline by gavage for 7 days,while dexamethasone was given with an intraperitoneal injection as a positive control.The remaining groups were pretreated with matrine and lycopene or their combination by gavage for 7 days.The dose of drug was dexamethasone(5mg/kg),matrine(25mg/kg),lycopene(1mg/kg),matrine(25mg/kg)+ lycopene(1mg/kg)group.After receiving an intra-gastric administration of normal saline or drugs daily for seven days prior to the administration of LPS(100μg/50μl)by trachea,except NM group administrated with normal saline.The drug treatment groups were given medication every day after giving LPS half an hour,and at six hours,one day,three days and seven days after LPS administration,all animals were anesthetized by chloral hydrate and sacrificed.Thereafter,lung tissues were collected for the analysis of the W/D ratio to assess tissue edema;the right lower lung was excised for histological examination;the expression of TNF-α and IL-6 were measured by ELISA and PCR and so on.We detected the levels of methane dicarboxylic(MDA),myeloperoxidase(MPO),and glutathione(GSH).Results: Compared with normal control group,LPS group’s W/D ratio markly increased,however,combinatorial treatment attenuated the LPS-induced W/D ratio increasing(6h 2.04±0.35 vs 2.85±0.38,1d 2.03±0.04 vs 3.07±0.12,3d 1.97±0.26 vs 2.78±0.06),(P<0.05).Meanwhile,histological examinations revealed protective effect of the combination against LPS-induced lung injury(6h 0.59±0.04 vs 1.05±0.07,1d 0.57±0.11 vs 1.22±0.05,3d 0.59±0.11 vs 1.18±0.11,7d 0.64±0.25 vs 1.01±0.10),(P<0.05).Combinatorial treatment also showed a substantial reduction in IL-6(ng/L)(6h 17.68±1.51 vs 25.32±0.73,1d 19.63±2.40 vs 29.60±2.42,3d 18.74±2.40 vs 23.98±1.40,7d 16.53±1.71 vs 20.69±0.61),(P<0.05)and TNF-α(ng/L)(6h 141.06±19.01 vs 197.19±22.83,1d 184.07±9.05 vs 231.85±7.56,3d 107.26±6.27 vs 161.32±16.36,7d 123.72±8.79 vs 159.60±11.55),(P<0.05)cytokines in lung tissue.RT-PCR analysis revealed increased m RNA expression of IL-6(6h 3.14±0.29 vs 4.06±0.19,1d 3.42±0.20 vs 4.43±0.13,3d 2.63±0.28 vs 3.45±0.32,7d 2.49±0.05 vs 3.32±0.26),(P<0.05),TNF-α(6h 2.52±0.15 vs 3.29±0.03,1d 2.67±0.18 vs 3.40±0.18,3d 2.39±0.09 vs 2.98±0.06,7d 2.13±0.14 vs 2.80±0.11),(P<0.05)and nuclear factor-kappa B(NF-κB)(6h 4.50±2.12 vs 9.50±0.71,1d 6.15±0.21 vs 11.00±1.41,3d 6.15±0.35 vs 10.50±2.12,7d 5.80±0.28 vs 9.00±1.41),(P<0.05)in LPS group,which were reduced by the combinatorial treatment.Additionally,to evaluate the anti-inflammatory activity of the combination of oxymatrine and lycopene by immunohistochemistry,the expression of pro-inflammatory cytokines NF-κB was studied.The results in the present study indicated that combinatorial treatment attenuated the expression of NF-κB(6h 4.50±2.12 vs 9.50±0.71,1d 6.15±0.21 vs 11.00±1.41,3d 6.15±0.35 vs 10.50±2.12,7d 5.80±0.28 vs 9.00±1.41),(P<0.05).We observed that mice exposed to LPS showed increased MDA(nmol/mg)(6h 11.33±0.38 vs 16.43±1.08,1d 14.78±0.38 vs 18.26±0.64,3d 13.05±0.15 vs 16.65±0.30,7d 10.28±1.58 vs 14.25±0.15),(P<0.05),and MPO(U/g)(6h 8.15±1.35 vs 9.75±0.47,1d 8.07±0.80 vs 9.66±0.44,3d 8.49±0.44 vs 10.00±0.24,7d 7.84±0.56 vs 10.04±0.21),(P<0.05)activity but marked decrease in GSH(mg/L)(6h 25.35±2.92 vs 18.41±2.08,1d 26.55±3.86 vs 17.17±2.51,3d 26.85±0.80 vs 17.12±1.16,7d 28.07±1.61 vs 20.61±1.03),(P<0.05)content.These changes were significantly reversed by treatment with the combination of oxymatrine and lycopene.Conclusion: Matrine combined with lycopene have protective effect on LPS-induced ALI,the protective effect of combinatorial treatment is to reduce the production of inflammatory factors and inhibit oxidative stress,its mechanism may be via suppression of NF-κB signal pathways. |
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