The Experimental Study On The Effect Of Xiaoshengsan Granules On TLR4 And NF-?B In Keratoconjunctiva Of Dry Eye Rat Model

Objective:1.Inhibitory Effect of Xiaosheng San Granules on TLR4 and NF-?B in Keratocon-junctiva of Mouse Dry Eye Model.2.Explore the effect of Xiaosheng San Granules for the treatment of dry eye through immuno-inflammatory inhibition.Methods:120 healthy C57BL/6 female rat that weighing between 20g and 25g,were randomized into 6 groups,that is normal group,model group,negative control group,low dose group,middle dose group,high dose group,20 in each group._In addition to the normal group,rat in other groups were,injected subcutaneously with scopolamine hydrobromide to establish an experimental dry eye model.In each group,the SIT,BUT and FL were measured on the 7th,14th,and 21st day of treatment,The SIT,BUT and FL were measured in the six groups on the 7th,14th,and 21th days of treatment.After each test,the rat were killed and the keratoconjunctival tissues were harvested for HE staining,immunohistochemistry,and Western blotting.Results:In therapeutic doses,when A treated with C57BL/6 rat for 7d?14d?21d,the therapeutic effects of the middle and high dose groups were statistically different from those of the model group(p<0.05).However,there was no significant difference in efficacy between the middle-dose group and the high-dose group(p>0.05),and there was a significant difference in efficacy between the low-dose group and the middle-dose group(p<0.05).There was no significant difference in SIT between the low dose group of Xiaoshengsan granules and the model group after 7d and 14d treatment(p>0.05).and after 7d?14d?21d treatment,compared with model group,BUT and FL are statistically different(p<0.05).The results showed that the middle and high doses of Xiaoshengsan was effective in treating dry eye,but the effect of low dose was not obvious,and the negative control group was ineffective.In treatment time,there was no statistically significant difference in SIT,BUT,and FL between the normal group,the model group,and the negative control group after 7 days,14 days,and 21 days of treatment(p>0.05).Significant differences in SIT,BUT,and FL were observed after 7d,14d,and 21 d in the medium and high dose groups(p<0.05),and with the extension of treatment time,the drug's effect becomes more pronounced.There was no significant difference in SIT between 7 days and 14 days in the Xiaoshengsan low dose group(p>0.05).There was no significant difference in BUT between the 7th,14th,and 21th days of treatment(p>0.05),and FL value was statistically significant(p<0.05).The results showed that the treatment effect of the middle and high dose groups became more obvious with the prolongation of medication time,the best effect was seen in 21 days,while the difference in time between low dose SIT and BUT was not obvious.HE staining showed that the keratoconjunctival tissue of the middle dose group of Xiaoshengsan Granules increased over time as the epithelial cells repaired hyperplasia,and the arrangement became tight and regular,until the 21d keratoconjunctival cells gradually returned to normal and arranged neatly and compactly.Immunohistochemistry of TLR4 and NF-?B showed strong positive expression on membranes of keratoconjunctival cells in the model group and the negative control group,which was expressed on the 7th day after treatment,decreased on the 14th day,and showed no positive expression on the 21st day(p<0.05).Western blotting result display the expression of TLR4 and NF-?B protein was statistically significant at 7d,14d,and 21d after treatment(p<0.05),the protein expression was gradually decreased with the prolonged administration time,and the protein expression was lowest at 21d.Conclusion:1.The use of scopolamine in conjunction with air blow induced C57BL/6 rat to establish a dry eye model has a high success rate and has a pathological process similar to humans.2.TRL4 and NF-?B were activated in a rat dry eye model and accelerated the development of dry eye in mice.Xiaoshengsan Granules treatment can effectively inhibit the activity of TRL4 and NF-?B,and the efficacy of 21st day in the middle and high dose groups is optimal.3.Xiaoshengsan Granules reduce the release of immune inflammatory factors by inhibiting the activity of TRL4 and NF-?B,further stabilizing the tear film and achieving the purpose of treating dry eye.