The Mechanism of Qingxuan Runmu Yin(清眩润目饮)on Benzalkonium Chloride-Induced Dry Eye in Rats Based on TLR4/MyD88/NF-κB Signaling Pathway
Objective:To evaluate the mechanism of Qingxuan Runmu Yin in the benzalkonium chloride(BAC)-induced dry eye model in rats based on the TLR4/MyD88/NF-κB signaling pathway.Methods:Totally 40 SD rats were randomely divided into blank group,model group,Qingxuan Runmu Yin(QRY)group and sodium hyaluronate group,with 10 rats(20 eyes)in each group.0.2％BAC eye drops were administered to each group of rats excluding the blank group,5 μL/eye,twice a day for 14 days,to prepare a dry eye model of rats with excessive evaporation.After modeling,the rats in the QRY group were given QRY by gavage and phosphate buffer drops.The rats in the sodium hyaluronate group were given equal amounts of saline by gavage and sodium hyaluronate eye drops.The rats in the blank and model groups were administrated with equal amounts of saline and phosphate buffers twice a day for a 14 days.Schirmer Test(SIT)and corneal sodium fluorescein(FL)staining scores of rats in each group were detected 1 hour after the last administration.Transmission electron microscopy was used to examine the ultrastructure of corneal cells.HE staining was performed to view the histopathologic changes in the conjunctiva.RT-qPCR was used to detect the expression of TLR4 mRNA,MyD88 mRNA,NF-κB p65 mRNA in the cornea,conjunctiva and lachrymal glands of rats in each group.ELISA was used to detect the expression of phosphorylated p-p65 protein in the cornea,conjunctiva and lacrimal gland of rats.Results:After 14 days of modeling,model group,QRY group and sodium hyaluronate group showed lower SIT than blank group(P＜0.01),while higher FL score than blank group(P＜0.01).After 14 days of administration,QRY group and sodium hyaluronate group showed significantly higher SIT than model group(P＜0.01 or P＜0.05),while lower FL scores than model group(P＜0.05).Transmission electron microscopy showed that the corneal epithelial cell gap was widened and irregularly arranged in the model group,while the cell gap widening was reduced and more regularly arranged in the QRY group and Sodium hyaluronate group.And the corneal structure was improved in QRY group and Sodium hyaluronate group.HE staining showed that compared with the model group,the morphology and arrangement of conjunctival epithelial cells were restored and the number of cup-shaped cells increased in QRY group and Sodium hyaluronate group.The model group showed higher TI.R4 mRNA and MyD88 mRNA in the cornea,conjunctiva and lacrimal gland,than blank group(P＜0.01).The QRY group and sodium hyaluronate group showed lower TLR4 mRNA and MyD88 mRNA in the cornea,conjunctiva and lacrimal gland,than model group(P＜0.01).The QRY group showed lower TLR4 mRNA(cornea)and MyD88 mRNA(conjunctiva and lacrimal gland)than sodium hyaluronate group(P＜0.01).The model group showed higher expression of p-p65 protein in cornea,conjunctiva and lacrimal gland,than blank group(P＜0.01).The QRY and sodium hyaluronate group showed lower expression of p-p65 protein in cornea,conjunctiva and lacrimal gland,than model group(P＜0.01).The QRY group showed lower expression of p-p65 protein in cornea and conjunctiva,than sodium hyaluronate group(P＜0.05).Conclusion:Qingxuan Runmu Yin can alleviate dry eye symptoms and repair corneal and conjunctival damage in benzalkonium chloride(BAC)-induced dry eye model rats,and its mechanism may be realized by inhibiting the TLR4/MyD88/NF-κB signaling pathway.
万方数据
维普
