Efficacies And Mechanisems Of Nanodrops Based On HMGB1 As The Intervention Target To Promote The Repair Of Corneal Epithelium

Keratopathy has been the second leading cause of blindness,the corneal epithelial disease with high incidence,so it has great significance to research the development mechanism and new treatment measures for timely repair the cornea and maintenance its integrity^[1].In this study,nano-drops were using DG as excipient,then we evaluated its properties and safety.We investigated the changes of High Mobility Group Box1（HMGB1）and its receptor during the repair of corneal epithelial injury.The expression of IL-6 and other inflammatory factors in cornea and trigeminal nerve were observed.In addition,we studied the repair of epithelial cells in normal mice by regulating the HMGB1 signaling pathway.The above process was repeated in diabetes mice.It provides a new idea for the treatment of keratopathy.Methods:PALM-DG nano-drops were using palmatine as main medicine,dipotassium glycyrrhizinate as excipient.First,the encapsulation rate was measured.Antioxidant capacity and safety;then pathological examination of eye drops and eye absorption were performed C57BL/6J mice were divided into five groups,which were PBS,Solution,DG,PALM&DG and PALM-DG.Eye drops were given to each group at the same time after scraped corneal epithelial.Corneal epithelial sensitivity;the degree of corneal and corneal nerve repair were measured at specific time;HMGB1 and receptor proteins in the cornea and trigeminal nerve of mice were measured by Western Blot（WB）;the inflammatory factors such as IL-6 and IL-1βin cornea and trigeminal nerve of mice were detected and analyzed by ELISA.Diabetes model was established with streptozotocin（STZ）（Ⅰtype）.After the blood glucose concentration was measured,diabetic mice were divided into PBS group,Solution group,DG group,PALM&DG group and PALM-DG group.Each group was treated with eye drops after curettage of corneal epithelium.The sensitivity of corneal epithelium and the repair degree of cornea and corneal nerve in diabetic mice were measured.Western Blot was used to detect HMGB1 and its receptor proteins in the cornea and trigeminal nerve of diabetic mice.ELISA was used to detect inflammatory factors such as IL-6 and IL-1βin the cornea and trigeminal nerve of diabetic mice.Results:（1）The drug and auxiliary ratio was 1:15 of PALM-DG nano-drops,and the liquid was clear and bright,with a light yellow color.The encapsulation rate of drops was 99.007±0.15%.The results showed that the ocular irritation was very low and the safety was high;enhance ocular absorption of drugs compared to drug solutions.（2）The PALM-DG group compered with others can recovery corneal sensitivity in a short time;effectively promoted the repair of corneal nerve in mice.Western Blot results showed that the expressions of HMGB1 protein and RAGE protein in cornea and trigeminal of PALM-DG group was low,which effectively inhibited the expressions of HMGB1 protein and RAGE protein.The results of ELISA showed that the expressions of corneal and trigeminal inflammatory factors of PALM-DG group were lower than those in other groups.（3）The PALM-DG group compered with others can recovery corneal sensitivity in a short time;effectively promoted the repair of corneal nerve in diabetic mice.Western Blot results showed that the expressions of HMGB1 and RAGE in cornea and trigeminal of PALM-DG group was low,which effectively inhibited the expressions of HMGB1 and RAGE protein in diabetic mice.The results of ELISA showed that the expressions of corneal and trigeminal inflammatory factors of PALM-DG group were lower than those in other groups.Conclusion:PALM-DG nanodrops have high safety,which can effectively improve the ocular absorption of the insoluble drug palmatine,and improve the pharmacological effectiveness and utilization rate of palmatine.For normal and diabetic mice,it can improve the repair speed of corneal epithelial,promote corneal nerve repair and corneal sensitivity recovery.The HMGB1 pathway is a key pathway of corneal epithelial injury in mice.Inhibition of the expression of HMGB1 and receptor is the key to prevent and cure corneal epithelial injury in mice.The PALM-DG nano-micelle can regulate this pathway to effectively prevent and cure corneal epithelial injury in mice.