Clinical Effect Of Xiaoyao Powder On Dry Eye With Anxiety And Depression Patient And Its Mechanism On CaMKⅡβ Pathway

Objective:Clinical study:To observe the clinical effect of Xiaoyao Powder on dry eye disease with anxiety and depression.Experimental study:To investigate the effects of Xiaoyao Powder on tear secretion,corneal epithelial fluorescein staining.and depressive behavior in mice with depression and dry eye,to observe the effects on the expression levels of CaMKIIβ,P-ATF2,Cox2 and p-p38 in hippocampus,and try to explore the mechanism of Xiaoyao Powder on depression,depression+dry eye.Methods:Clinical study:Patients with dry eye disease and anxiety and depression from the Department of Ophthalmology,Jinhua Hospital of Traditional Chinese Medicine were randomly divided into two groups between September 2019 and August 2022.Experimental group received oral taken Xiaoyao Powder,combined with 0.3%sodium hyaluronate eye drop.Control group received 0.3%sodium hyaluronate eye drop only.The whole procedure was 1 month.The TCM syndrome scale was used to observe the changes of liver depression and qi stagnation syndrome,NRS was to evaluate pain,HAD was to evaluate anxiety and depression,SPEED was to evaluate dry eye symptoms,the TMH,tear secretion,BUT,corneal epithelial fluorescein staining.,and tear IL-1β level were observed before and after treatment.2.Experimental study:40 healthy,6-week SPF C57/BL6J female mice were randomly divided into 5 groups(normal group,dry eye group,depression group,dry eye and depression group,dry eye and depression and Xiaoyao Powder group).Each group has 8 mice.Experimental dry eye was induced in by subcutaneous injection of Scopolamine hydrobromide(Sigmae Aldrich)four times a day(08:00,11:00,14:00,and 17:00)with exposure to an air draft and 15%humidity,26℃,for 5 days.Reserpine was injected on the 6th day to induce depression model.The saline and Xiaoyao Powder were given by gavage administration.The corneal epithelial fluorescein staining.staining,tear secretion,the stationary time of treadmill test and tail suspension test were used to determine the depression state e and the effect of Xiaoyao Powder,and the level of P-ATF2,Cox2,p-p38 and CAMKⅡβ in the hippocampus were detected.Results:1.Clinical study:There were no significant difference in age,sex and course of disease between the two groups(P>0.05).① TCM syndrome score:before treatment:the value of TCM syndrome integral in the experimental group was 7.00(5.75,11.25),which was 7.50(5.75,10.00)in the control group.There was no significant difference between the two groups(Z=-0.064,P=0.949>0.05).After treatment:the TCM syndrome score in the experimental group was 2.50(2.00,4.00),which was 4.50(3.75,6.00)in the control group.There was a statistically significant difference between the two groups(Z=-3.834,P<0.001).②NRS:The NRS score was 3.00(2.75,5.00)in the experimental group and was 3.50(3.00,5.00)in the control group.There was no statistically significant between the two groups(Z=-1.192,P=0.233>0.05).After treatment:The NRS score 1.00(1.00,2.00)in the experimental group,which was 2.00(1.75,3.00)in the control group.There was a statistically significant difference between the two groups(Z=-4.128,P<0.001).③ SPEED:Before treatment:the SPEED in the experimental group was 16.00(13.00,19.75),which was 16.00(13.75,20.25)in the control group,there was no statistically significant between the two groups(Z=-0.108,P=0.914>0.05).After treatment:the SPEED in the experimental group was 7.50(4.00,10.00),which was 10.00(8.00,13.00)in the control group.There was a statistically significant difference between the two groups(Z=3.539,P<0.001).④HAD:Before treatment:the HAD in the experimental group was 12.00(8.00,19.25),and which was 12.00(9.75,16.25)in the control group.There was no statistically significant between the two groups(Z=-0.652,P=0.515>0.05).After treatment:the HAD in the experimental group was 6.00(4.75,8.25),which was 9.50(7.00,11.25)in the control group.There was a statistically significant difference between the two groups(Z=-3.068,P=0.002<0.05).⑤ BUT:Before treatment:the BUT in the trial group was 1.00(1.00,2.00),which was 1.00(1.00,2.00)in the control group.There was no statistically significant between the two groups(Z=-1.155,P=0.248>0.05).After treatment;the BUT in the trial group was 5.00(5.00,7.00),and was 5.00(3.75,5.00)in the control group.There was a statistically significant difference between the two groups(Z=-2.744,P=0.006<0.05).⑥Corneal epithelial fluorescein staining.score:Before treatment:the corneal epithelial fluorescein staining.score in the experimental group was 2.00(0.75,3.25),which was 2.00(1.00,3.25)in the control group.There was no statistically significant between the two groups(Z=-0.290,P=0.772>0.05).After treatment:the corneal epithelial fluorescein staining.score in the experimental group was 0.00(0.00,1.00),which was 1.00(0.00,1.25)in the control group.There was a statistically significant difference between the two groups(Z=-2.307,P=0.021<0.05).⑦TMH;Before treatment:The TMH was 0.15±0.02mm in the experimental group and was 0.14±0.01mm in the control group.There was no statistically significant between the two groups(t=1.953,P=0.060>0.05).After treatment:The TMH was 0.29±0.05 mm in the experimental group and was 0.24±0.03 mm in the control group.There was a statistically significant difference between the two groups(r=-4.312,P<0.001).⑧Tear secretion length:Before treatment:The tear secretion length in the experimental group was 6.80±2.24mm/20s,which was.6.50±1.50mm/20s in the control group.There was no statistically significant between the two groups(t=0.699,P=0.490>0.05).After treatment:The TMH was 14.50±1.31mm/20s in the experimental group and was 12.30±1.15mm/20s in the control group.There was a statistically significant difference between the two groups(t=7.047,P<0.001).⑨IL-1β in tear:Before treatment:The IL-1β level in the experimental group was 23.62±2.15 ng/L,which was 23.18±1.78 ng/L in the control group.There was no statistically significant between the two groups(t=1.086,P=0.286>0.05).After treatment:The IL-1β level in the experimental group was 19.28±1.36 ng/L,while it was 20.36±1.57 ng/L in the control group.There was a statistically significant difference between the two groups(t=-2.844,P=0.008<0.05).2.Experimental study:Compared with the normal group,the tear secretion in the dry eye group was significantly reduced,and the absolute value of the tear volume measured by hiphenol cotton thread was less than 2mm,indicating that the dry eye model was successfully established.Tear secretion:Compared with the normal group,the tear secretion in the depression group was significantly decreased(P<0.05);The tear secretion in dry eye+depression group was significantly lower than that in depression group and dry eye group(P<0.05).Compared with the dry eye and depression groups,the tear volume of Xiaoyao Powder treatment group was significantly increased(P<0.01).Corneal epithelial fluorescein staining:The dry eye group and dry eye+depression group were higher than that in the control group(P<0.05).Compared with the dry eye+depression group,the corneal epithelial fluorescein staining.in Xiaoyao Powder treatment group was significantly decreased(P<0.05).Behavioral evaluation:The results of fixed time after tail suspension showed that compared with the normal group,the immovable time of tail suspension was significantly increased in the depression group,dry eye+depression group,and dry eye+depression+treatment group(P<0.05).There was no significant difference between the dry eye group and the normal group(P>0.05).Compared with the dry eye+depression group,the tail hanging fixed time in the treatment group was significantly reduced(P<0.05).Compared with the normal group,the times of electric shocks in the depression group was significantly increased(P<0.05).Compared with the normal group,the times of electric shocks in the dry eye+depression group was significantly increased,and was higher than that in the depression group(P<0.05).After treatment with Xiaoyao Powder,the number of electric shocks was significantly decreased compared with the normal group(P<0.05).Brain tissue protein detection:the expression levels of P-ATF2,Cox2,p-p38,CAMKIIβ were significantly increased in the depression group and dry eye+depression group compared with the normal group,and them were significantly decreased after using of Xiaoyao Powder(P<0.05).Conclusions:The findings of clinical study show that the use of Xiaoyao Powder play a useful and positive role in dry eye disease with anxiety and depression patients.Xiaoyao Powder can can aid the recovery of corneal surface and promote the stability of tear film.Xiaoyao Powder can effectively promote the tear secretion,light ocular pain and relieve dry eye symptoms,promote tear film to be more stable,increase the TMH,and down-regulate the level of tear IL-1β.2.Experimental study:Depression would increase the risk of dry eye and deteriorates ocular surface performance in mice.Experimental studies show that depression increases the risk of dry eye in mice.Xiaoyao Powder has a significant therapeutic effect on dry eye with depression mice.Xiaoyao Powder can promote tear secretion,reduce corneal staining,and improve depression behavior in mice.The main findings of this study indicate Xiaoyao Powder can2 play a protective role in the cerebral neurons in mice.It may be related to the regulation of CAMKⅡβsignal pathway in the hippocampus,by down-regulating the expression levels of P-ATF2,Cox2,p-p38.It also indicate that the symptoms of ocular surface damage and behavior in dry ere disease with depression is related to dysfunction in cerebral neurons.