The Quality Control Of Zhenzhu Mingmu Drop Eys And Its Efficacy For Dry Eys Syndrome And Study On Mechanism

ObjectiveZhenzhu Mingmu Drop Eyes(ZMDE),is mainly composed of pearl and borneol,and its pharmacological actions is relative with the free amids acid in it.ZMDE has the effective of cleaning heat,nourishing liver and lighting eyes,chronic conjunctivitis and dry eys syndrome,and so on.Some amino acids have pharmacological effects of adjusting internal secretion and improving immunity,such as Serine,valine,and so on.Others have effects of delaying age,anti-inflamation and anti-oxigen,such as alanine,glycine,asp,leucine and aginine.Considering the delivering system and drug dynamic feature of ZMDE,the substances that have phamocologic effectives very likely are free amino acids,such as alanine,glycine,asparagic acid and arginine,and so on,which have the effectives of anti-imflamation and anti-oxygen,while bound amino acids that is the multi-peptides.Thus,it’s necessary to going on the research of quality control of free amino acids in ZMDE.In order to providing reference for the quality control of ZMDE,we established the PITC column-before deriviated High Performance Liquid Chromatography method to determine quantitily the 17 kinds of free amino acids nine different manufacture Zhenzhu Mingmu Drop Eyes preparation.The liver has relationship with eyes in Chinese medicine.The liquid of liver decrease,the tears secretion decreased accordingly;the gas of liver over,then the evaporation of tears increase.The ZMDE have effects of relieving the tired of eyesight,caused by the liver liquid lack and the liver gas overflow.The quantity of tear drop secretion decreased,causing the unstable of tear film,and the osmotic pressure of ocular surface up,strengthening the evaporation of ocular surface tear volume,the unstable of ocular surface tear film and the high evaporation pressure interaction,sustaining the nuclear mechanism of epithelium hurting of ocular surface,and causing the inflammation of dry eye disease of ocular surface.The syndrome occures including eyesight tired,conjunctiva over blood and conjunctiva dye color,as the dry eye happened.This research was designed to explore the the quality control of ZMDE and its curing dye eyes,and reveal the mechanism of dry eye disease,by analysising the research results of ZMDE curing for the model of dry eye,made by atropine sulfate.Methods1.Derived with phenyl isothiocyanate(PITC),then analyzed by high performance liquid chromatography(HPLC)method.The samples were analyzed on a Venusil-AA amino acid analysis column(250 mmX4.6 mm,5μL)with gradient elution of 0.1 moL·L-1 sodiumacetate solution(adjusted to pH 6.5)-acetonitrile(93:7)(A)and 80%acetonitrile(ultrapure water account for the 20%volume)(B)at the flow rate of 1.0 mL·min-1.The detected wavelength was 254 nm,and the column temperature was 38℃.2.In order to study the function mechanism of ZMDE curing for dry eye syndrome,the Schirmer-II method was adopted to test the rabbit tear secretion under differ statement.The conjunctival staining method was adopted to test the hurt level of conjunctival epithelial cells.The Ferning test was adopted to test the dry eye level of ocular surface.The ELISA method was adopted to test the inflammation level of conjunctival cells.Results1.Determine and analysis the most part amino acids substances were glycine,arginine,tyrosine and and leucine from different manufactures ZMDE,by HPLC method,providing that the content and stable of four amino acids above might be the key point of quality control of ZMDE.2.Discovered that the basic effective substance curing for dry eye was pearl liquid or pearl-level powder liquid from ZMDE.The ocular surface tear secretion caused by the different concentration ZMDE,was different obviously.Conclusion1.The ZMDEs from different manufactures have multi-kinds amino acids,the glycine,arginine,tyrosine and leucine is the key point of quality control of ZMDEs.2.By adjusting ocular surface tear secretion and enhancing the anti-oxigen hurt ability of conjunctiva of ocular surface,improve ocular surface dry eye condition.The effective basic substance is pearl liquid,which linked with the arginine in pearl liquid,most likely.