| Objective:In this study,a rabbit model of dry eye with water deficiency was established by subcutaneous inj ection of scopolamine hydrobromide,and Qingxuan Runmu Yin was given to intervene.The effects of Qingxuan Runmu Yin on Schirmer I test,tear film rupture time,cornea and conjunctival tissue morphology,and the expression changes of IL-1beta,IL-6 and TNF-alpha protein in cornea and conjunctiva were analyzed to study Qingxuan Runmu Yin.The therapeutic effect and mechanism of Qingxuan Runmu Yin on rabbit dry eyes with water deficiency type.Its mechanism of action on rabbit dry eyes with water deficiency.Methods:Thirty male healthy Japanese big-eared white rabbits(weighing 2.0 ± 0.5kg)were randomly divided into six groups:normal group,model group,western medicine control group,high,medium and low dose groups of Chinese medicine.Normal group did not do any treatment,the other groups were subcutaneously inj ected with scopolamine hydrobromide to replicate animal models(4 mg/rat,4 times/day,14 days after intervention).The high,middle and low dose groups of Chinese medicine were given Qingxuan Runmu Yin by gavage at doses of 19.40g/kg,9.70g/kg and 4.85g/kg.Normal group and model group were given normal saline once a day.Western medicine control group was given tobramycin and dexamethasone eye drops combined with sodium hyaluronate artificial tear eye drops,three times a day,point eyes,each group was given 28 days continuously.Schirmer I test(SIt)and break-up time(BUT)were measured in all rabbits on the 1 st day before,14th day after,14th day after and 28 th day of administration.On the 29th day after administration,all experimental animals were killed by excessive anesthesia,and corneal and conjunctival tissue sections were prepared by hematoxylin-eosin staining.The morphological changes of cornea and conjunctiva were observed by HE staining.The concentrations of IL-1beta,IL-6 and TNF-alpha in cornea and conjunctiva were detected by ELISA.Results:1.SIT:Compared with the model group,there were significant differences in the 14th day and 28th day(P<0.05).There was no significant difference between the high dose group and the normal group on the 28th day(P>0.05).There was no difference between the low dose group and the model group on the 14th day of administration(P>0.05),and there was significant difference between the model group and the low dose group on the 28th day of administration(P<0.05).2.BUT:Compared with the model group,there were significant differences in the 14th day and 28th day(P<0.05).There was no difference between the low dose group and the model group on the 14th day of administration(P>0.05),and there was significant difference between the model group and the low dose group on the 28th day of administration(P<0.05).3.IL-1beta,IL-6,TNF-alpha:The expressions of IL-1beta and IL-6 in the model group were not significantly different from those in the low dose group(P>0.05),and there were significant differences compared with other groups(P<0.05).Two or two comparisons were made among the four groups treated:there was significant difference between the low-dose group and the high-dose group and the western medicine control group(P<0.05);there was no significant difference between the two comparisons among the other groups(P>0.05).The content of TNF-a in the model group was different from that in the other five groups(P<0.05),and there was no significant difference between the four groups treated(P>0.05).4.HE staining:Compared with the normal group,the corneal epithelial cells in the model group were exfoliated and absent,and gradually became squamous epithelialization near the corneal surface,with edema in the stroma.The conjunctival epithelial cells were irregularly arranged,and goblet cells were significantly reduced,showing a large number of inflammatory cells infiltration.The morphology of cornea and conjunctiva of rabbits in western medicine control group and high dose group of Chinese medicine was similar to that of normal group,and epithelial cells arranged regularly;conjunctival goblet cells were abundant in epithelial cells,with a small amount of inflammatory cells infiltrating,which showed a trend of weakening pathological changes as a whole.Conclusion:1.The model preparation method of hypodermic injection of scopolamine hydrobromide can induce experimental immune inflammation in rabbits and successfully establish a water-deficient dry eye animal model.2.Qingxuan Runmu Yin can increase tear secretion,prolong tear film rupture time,and promote the repair of dry rabbit corners and conjunctival epithelial cells with water deficiency.3.Qingxuan Runmu Yin can down-regulate the expression of IL-1beta,IL-6 and TNF-alpha in rabbit horn and conjunctiva.4.The mechanism of Qingxuan Runmu Yin’s "Pulmonary-Spleen Therapy" may be related to immune regulation.By down-regulating the expression of IL-1beta,IL-6 and TNF-alpha inflammatory factors,it can interfere with the immune inflammation reaction of water-deficient dry eyes,promote the increase of tear secretion,prolong the tear film rupture time,accelerate the repair of corneal and conjunctival epithelial cells,and further achieve the purpose of treating water-deficient dry eyes. |
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