| Dry eye is one of the very common clinic diseases,the main clinical manifestations are dry eye,foreign body sensation,photophobia,eyelids tension and discomfort,severe cases can cause visual function decline,thereby affecting the work and life.Chinese medicine believes that "the liver opens into the eyes","the liver in the liquid for the tears",the subtle material of the liver is stored in the eyes,so that the eyes are nourished,so as to maintain the normal visual function.Xiaosheng San is the clinical experience by mentor XieLike,commonly used in the clinical treatment of patients with dry eye syndrome is stagnation of liver depression and yin deficiency.Use dredging liver and nourishing Yin Syndrome Differentiation and treatment of dry eye,early application in clinical and animal experiments have shown some efficacy,this study aims to further explore the treatment effect of Xiaosheng San to dry eye and molecular mechanism,provide a new idea and an objective basis for TCM treatment of dry eye.ObjectiveTo observe the distribution of stagnation with liver depression and yin deficiency type of dry eye syndromes elements;differences and correlation between measuring three different inspection methods do tear film stability;observe tear film stability,clinical symptom scores,SchimerI test and comeal fluorescein staining before and after Xiaosheng San treatment to liver depression and yin deficiency type of dry eye;the role of stem cell injury model of corneal epithelium,to further explore the effect and molecular mechanism of Xiaosheng San treatment of dry eye.Method1 According to the diagnostic criteria of dry eye and the standards of TCM outpatients,in our hospital 100 cases of dry eye syndrome in patients with depression due to liver depression and yin deficiency,the TCM syndrome items are divided into no,mild,moderate and severe grade 4,respectively to 0,1,2,3 points,interrogation accurate records of patient imformation,statistical analysis according to the integral method of stagnation of liver depression and yin deficiency type of dry eye clinical research.2 Sirius,RT-7000 and FLS were used to measure the tear film stability of dry eye patient,and the difference and correlation between the three methods were analyzed and compared3 Outpatients in our hospital 80 cases of dry eye syndrome in patients with liver depression and yin deficiency,oral medicine Xiaosheng San treatment for 4 consecutive weeks,before and after treatment,observe the change of tear film stability,clinical symptom scores,SchimerI test and corneal fluorescein staining.4 Human corneal epithelial cells joined Benzalkonium Chloride Solution prepared dry corneal epithelial cell injury model,adding low,medium and high dose of Xiaosheng San serum and control serum containing blank after incubation,respectively in 24h,48h,72h function by light microscopy to observe the morphologic changes of comeal epithelial cells,cell viability was determined with CCK-8 method,detect the expression corneal epithelial cell apoptosis was detected in RT-PCR and IL-1,IL-6,TNF-a,MMP-9 cytokine flow cytometry.Result1 This study included patients with an average age of 42.09,the minimum 18 years,maximum 60 years old;male sex,44 people(44%),56 female(56%);occupation,14 workers(14%),retired 9(9%),5(accounting for the unemployed 5%),4 students(4%),staff of 68 people(68%);syndrome elements(except tongue and pulse)proportion from big to small are the top three,dry eyes,tingling,irritability.2 Rupture time difference between the three methods for measuring the tear film was statistically significant(P<0.05),comparison results showed that the FLS method for the measurement of the break-up time is relatively long compared with Sirius and RT7000,Sirius and RT7000 and the break-up time measurement is relatively short and no difference between them.Pearson correlation analysis showed that the correlation coefficient of FLS and Sirius was 0.39,the correlation coefficient of FLS and RT7000 was0.41,statistically significant(P<0.05).3 After taking Xiaosheng San,the break-up time of tear film measured by three methods was longer,and the time was statistically significant(P<0.0001),The results of correlation analysis show that the measurements of same method before and after taking traditional Chinese medicine were in high consistence,correlation coefficient of Sirius method before and after medication reached 0.81(P<0.0001),the correlation coefficient of RT7000 was 0.83(P<0.0001),and the measurement results of FLS consistency is poor,the correlation coefficient was 0.68(P<0.0001);clinical symptom scores decreased,tear secretion increased and corneal fluorescein staining declines after treatment,and all results were statistically significant(P<0.0001).4 With benzene as chloramines concentration lug/ml,low toxicity on corneal epithelial cells,the cells with time slowly,it vacuoles gradually increased,cell activity is relatively low,moderate the effect,the choice of BAC lug/ml as a model of concentration concentration.The serum concentration of not more than 10%,had no effect on the activity of corneal epithelial cell,so the selection of 10%blank serum as the control,2.5%,5%,10%of Xiaosheng San medicated serum as traditional Chinese medicine,low middle and high dose group experiment.In the traditional Chinese medicine,the high dose group intervention corneal epithelial cell injury model,can inhibit the increase of cell volume,cell lysis and bigger space,reduce the cell wall and the removal of dissolved and necrosis;apoptosis;cell activity was detected by CCK-8 assay.The cell activity of Chinese herbal medicine and high doses of the value and the other group differences were statistically significant(P<0.05);RT PCR,IL-6,TNF-a and IL-1 to detect the expression of MMP-9.The results show that the relatively high in traditional Chinese medicine and the high dose group and the results showed with the increase of drug concentration,the inhibitory effect of relatively better results,24h,48h and 72h were low point in the traditional Chinese medicine,the high dose group of inhibiting inflammation the effects were more obvious(P<0.05).Conclusion1 Liver depression and yin deficiency type of dry eye patients in the majority of women,usually at around 40 years old,often engaged in the occupation for staff,workers,retirees,the main symptoms are dry,tingling,irritability.2 Measurement with three kinds of different examination methods of dry eye tear is different,FLS method for measuring a relatively long period of time,Sirius and RT7000 measurements over a relatively short period of time,but the three method of measuring the break-up time consistency in the trend,for the clinical diagnosis and treatment of dry eye has certain significance.3 After taking Xiaosheng San for liver depression and yin deficiency type of dry eye,the break-up time of tear film measured by three methods was longer,and the measurements of same method before and after taking traditional Chinese medicine were in high consistence;clinical symptom scores decreased,tear secretion increased and corneal fluorescein staining declines after treatment,further confirmed the effection of Xiaosheng San treatment.4 Xiaosheng San medicated serum medium and high dose group could significantly increase cell activity;apoptosis;Xiaosheng San could decrease the production of IL-1,thereby reducing the production of IL-6,TNF-a and other inflammatory factors as well as possible by reducing the production of IL-1,the number and activity regulation of MMP-9,play a protective role in corneal epithelial cells. |
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