ABSTRACT
The present study was designed to establish a suitable assay to explore CCR2b receptor antagonists from the natural products of Artemisia rupetris and Leontopodium leontopodioides. An aequorin assay was developed as a cell-based assay suitable for 384-well microplate and used for screening CCR2b receptor antagonists from natural products. Through establishing suitable conditions, the assay was shown to be suitable for screening of CCR2b receptor antagonists. Seven compounds were identified in preliminary screening. Five of them showed evident dose-response relationship in secondary screening. The structure-activity relationship study suggested that 7-position hydroxyl group of flavonoids was necessary, a polar group should be introduced on the 3-position, and the substituents on 2-position benzene ring of flavonoids have little influence on the potentency of the inhibition activity on CCR2b receptor. The ortho-position dihydroxyl structure in quinic acid compounds may be important. In conclusion, Compounds HR-1, 5, 7, and AR-20, 35 showed activity as antagonist of CCR2b receptor, which shed lights on the development of novel drugs as CCR2b receptor antagonists for preventing inflammation related diseases.
Subject(s)
Humans , Artemisia , Chemistry , Asteraceae , Chemistry , Drug Evaluation, Preclinical , Kinetics , Plant Extracts , Chemistry , Pharmacology , Receptors, CCR2 , Genetics , Metabolism , Structure-Activity RelationshipABSTRACT
Background Age-related cataract is the leading cause of visual impairment worldwide.To seek the effective prevention and drugs for management of cataract is important.Naphthalene-induced cataract of rat is an ideal animal model for the research of human age-related cataract,and aspirin has been proven to inhibit the development of human age-related cataract.ObjectiveThe present study is to investigate the role of aspirin on naphthalene-induced cataract.Methods Forty-five 150-160 g female SD rats were divided into three groups randomly.Naphthalene was orally taken with 0.5mg/kg per day for 3 days and then 1mg/kg per day for 70 days,and then 100mg/kg of aspirin was given per day for 70 days following four-day washout period in group A.In group B,the animals was given orally only naphthalene at the same way.No any intervention was used in group C.Naphthalene-induced cataract was examined under the slim lamp every week.The experimental animals were sacrificed and lenses were obtained in 70 days.α-Crystalline was extracted from lens homogenate and purified and identified using High Performance Liquid Chromatography(HPLC),2-dimentional electrophoresis gel and Western blot.Different abilities of α-crystalline to protect β low crystalline from aggregation were observed using ultraviolet spectrophotometer.Results Naphthalene-induced cataract formed at the third week in only naphthalene group but at the sixth week in naphthalene+aspirin group under the slim lamp.No significant difference was found in the degree of lenses opacity in the second week among these three groups(F=0.032,P=0.969).However,a statistically significant difference was seen in the degree of lenses opacity in the fourth,sixth,eighth and tenth week among these three groups(F= 5031.130,P=0.000;F=115964.000,P=0.000;F=169846.500,P=0.000;F= 195431.200,P=0.000).Themolecular chaperone-like activity was significantly higher than that of the naphthalene-induced group.Conclusion Aspirin delays the progression of lens opacification through protecting α-crystalline molecular chaperone-like activity.