Effects of Ophiopogon japonicus polysaccharides on inflammatory response in mice with Aspergillus fumigatus keratitis
Objective To investigate the therapeutic effect and its mechanism of Ophiopogon japonicus polysaccharides(OJP)on Aspergillus fumigatus(AF)-induced keratitis(AFK)in mice.Methods The animal model of AFK was established by injecting AF suspension into the cornea of mice.Fifty-four AFK mice were randomly divided into 5 groups:model group(n=11),low dose group(n=11),medium dose group(n=11),high dose group(n=11)and agonist group(n=10).The unmodeled mice were selected as control group(n=10).Mice in low dose group,medium dose group and high dose group were given 50 μL eyedrops of 5,10,20 g/L OJP solution.Mice in agonist group were given 50 μL eyedrops of 20 g/L OJP solution and gavaged with 0.25 mg/kg Toll-like receptor 4(TLR4)activator CRX-527 solution.Mice in control group and model group were given 50 μL normal saline as eyedrops.All mice in each group were treated for one week.At 24 h after treatment,the corneal inflammation was scored under slit lamp microscope.Corneal morphology was observed by hematoxylin eosin(HE)staining.The levels of pro-inflammatory factors interleukin(IL)-1β,tumor necrosis factor-α(TNF-α)and IL-6 in the supernatant of corneal tissue were detected by ELISA method.The mRNA levels of IL-1β,TNF-α,IL-6,TLR4,myeloid differentiation factor 88(MyD88)and nuclear factor kappa B(NF-κB)p65 in cornea were analyzed by qRT-PCR.The relative levels of TLR4,MyD88 and NF-κB p65(nucleus)proteins in cornea were analyzed by Western blot.Results Compared with control group,the corneal inflammation score was increased in model group(P＜0.05),the corneal lesions were obvious,the protein and mRNA levels of IL-1β,TNF-α,IL-6 in corneal were increased(P＜0.05),and the mRNA and protein levels of TLR4,MyD88,NF-κB p65 in corneal were increased(P＜0.05).Compared with model group,the corneal inflammation score was decreased in low dose group,medium dose group and high dose group(P＜0.05),the corneal lesions were improved,the protein and mRNA levels of IL-1β,TNF-α,IL-6 in corneal were decreased(P＜0.05),and the mRNA and protein levels of TLR4,MyD88,NF-κB p65 in corneal were decreased(P＜0.05).Compared with low dose group,the corneal inflammation score was decreased in medium dose group and high dose group(P＜0.05),the corneal lesions were alleviated,the protein and mRNA levels of IL-1 β,TNF-α,IL-6 in corneal were decreased(P＜0.05),and the mRNA and protein levels of TLR4,MyD88,NF-κB p65 in corneal were decreased(P＜0.05).Compared with high dose group,the corneal inflammation score was increased in agonist group(P＜0.05),the corneal lesions were worsened,the protein and mRNA levels of IL-1 β,TNF-α,IL-6 in corneal were increased(P＜0.05),and the mRNA and protein levels of TLR4,MyD88,NF-κB p65 in corneal were increased(P＜0.05).Conclusion OJP can partly attenuate the corneal inflam-matory injury in AFK mice by inhibiting TLR4/MyD88/NF-KB signaling pathway.
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