Ginkgo Biloba Extract On Interleukin-1¦Â Effects And Mechanism Of Injury In Rat Articular Chondrocytes,

Objective To observe the protective effects of Ginkgo Biloba Extract (EGb761) on damage chondrocytes of knee-joint injuried by rratIL-1β in rats and to explore the protective mechanisms of EGb761 through free radicle, nitric oxide, inducible nitric oxide synthase, NF - k B Signal Transduction pathway, matrix metalloproteinase-13. Try to investigate a better therapeutic approach and medicines for osteoarthritis.Methods (l)Culture and Identification of Rat Chondrocyte: Cartilaginous layers of knee were separated from one month' s old Wistar rats under semi-asepsis environment and then digested under sterile environment to get cartilaginous cells. The experimental cells were the second generation cartilaginous cells, which identified with extracellular matrix in culture. These cells were observed by inverted microscope and then were taken photos. (2)Chondrocyte were divided randomly into eight groups cultured with different cultures: rratIL-1β ( 5 μ g/L ) group, rratIL-1β( 5 μ g/L ) with different concentrations of dexamethasone (10^-7 10^-6 10^-5mmol/L) groups, rrat IL-1β ( 5 μ g/L ) with EGb761( 40mg, 80mg, 160mg/L)groups and control group. After 72hours, the chondrocyte multiplication were detected by MTT assay and total protein were measured by Coomassie Brilliant blue G-250. (3)Chondrocyte were divided randomly into the same eight groups. After 24 hours, cul ture medium was removed. SOD and iNOS' s activities were detected by chemical colorimetry method. NO was measured by methid of nitrifying ferment, inducible Nitric Oxide Synthas (iNOS) mRNA content was measured by RT-PCR method. The aboving methods were used to observe the effect of EGb761 on NO, iNOS activity, iNOSmRNA, SOD of damaged chondrocyte injuried by rratIL-1 β . (4) Chondrocyte were divided randomly into six groups cultured with different cultures: rratIL-1 β( 5 μ g/L ) group, rratIL-1 β ( 5 μ g/L ) with concentrations of dexamethasone (10^-6 mmol/L) group, rrat IL-1 β( 5μg/L )with EGb761( 40mg , 80mg, 160mg/L) groups and control group. NF-k B p⁶⁵ activity and rat matrix metalloproteinase (MMP-13) content were detected by ELISA assay.Results (l)The phaenotype of chondrocytes was metachromasia in...