Osteoarthritis-i Side The Efficacy Of The Treatment Of Osteoarthritis And Cartilage Cell Proliferation And Apoptosis Gene Expression, Anti-injury Research

objective : To observe the effect of ostearthritis fangâ–¡on ostearthritis;to observe the effect of osteoarthritis fangâ–¡on cultured chondrocyte proliferation in vitro; to observe the effect of osteoarthritis fangâ–¡on the regulate for Fas, Bcl-2 gene expression and on against hydrocortisone damage and on the recovery ability to chondrocyte DNA leision caused by H₂O₂ on cultured chondrocyte in vitro.methods: (1) observation of clinical therapeutic effect. osteoarthritis fangâ–¡ was made for precompounded prescription order, everyday dosis, was decocted in water for oral dose。 30 patiens were observed and were treated for 2-4 weeks,who accorded with OA diagnostic criteria of American rheumatology institute in 1995.the therapeutic effect standard were evaluated by referencing to new drug development guide of the traditional Chinese medicine for OA curative effect standard in 2002. (2) empirical study: the chondrocyte was isolated from cartilage of 3-4 weeks newly born rabbit by means of collagenaseâ–¡digestion and identified by image analysis,toluidine blue staining , alcian bllie staining,etc.Different dose of osteoarthritis fangâ–¡were added to the chondrocyte and incubuted .The effects of osteoarthritis fangâ–¡on the proliferation of chondrocyte was monitored by MTT analysis; the osteoarthritis fang I were added to the chondrocyte and incubated,contrasting to phosphate buffered solution,the Fas, Bcl-2 geneexpression of chondrocyte were detected through flow cytometry in order to evaluate the role to regulate apoptosis gene; the mixed liquor of osteoarthritis fang I and hydrocortisone were added to the chondrocyte and incubuted . Morphologic changes of chondrocyte were observed through inverted microscope.and the effect against hydrocortisone damage were monitored by MTT analysis; the mixed liquor of osteoarthritis fang I and H2O2 were added to the chondrocyte and incubuted .the frequency of cometal cells and variation of comet tail length were monitored by single cell gel electrophoresis analysis in order to approach the recovery ability to chondrocyte DNA leision caused by H2O2 on cultured chondrocyte in vitro.result: clinical therapeutic effect: the OA symptoms on paku swelling ^ early morning stiffness > tenderness etc were improved obviously by treated osteoarthritis fangdi. above all, the curative effect of early morning stiffness surpass control group; experimental result: K chondrocyte was identified by toluidine blue staining and atto-fresh-blue staining.2-. 5% and 2.5% of osteoarthritis fang LI obviously promoted for chondrocyte proliferation for 48 hour on cultured.howover ,it was no significative discrepancy between 0. 5% of osteoarthritis fang I and control experiment group .3 -, the 7.5% concentration of osteoarthritis fang I was added to the chondrocyte and incubated,which remarkably promoted FasN Bcl-2 genetic expression of cartilage cell on cultivation for 4 hour, comparing to the controlexperiment group . 4 N From observing cartilage cell Morphologic change, 1.25mg/mk 2.5mg/ml simple concentration of hydrocortisone almostly made chondrocyte die on cultivation in vitro for 24 hour; however,the mixed liquor of 1.25mg/ml concentration of hydrocortisone and 7.5% concentration of osteoarthritis fang I were added to the chondrocyte and incubated, the cell growed well; the mixed liquor of 2.5mg'ml concentration of hydrocortisone and 15% concentration of osteoarthritis fang I were added to the chondrocyte and incubated, the cell growed well,too; the result of MTT analysis accorded with cartilage cell morphous observation. 4^ the frequency of cometal cells : the frequency of cometal cells in the group of H2 02 was superior to the mixed group of H2 O2 and osteoarthritis fang I N osteoarthritis fang I and control group (P<0.01 ) . .It was significant difference between the osteoarthritis fang I and control group. The variation of comet tail length : the image of the DNA fluorescence was rotundity in the the osteoarthritis fang I and control group, the image of the cometic DNA fluorescence appear ed in the H2O2 group and the mixed group of H2O2 and osteoarthritis fang I .however,the comet tail in the he mixed group of H2O2 and osteoarthritis fang I was shorter than the H2O2 group( P<0.01 ). conclusion: the osteoarthritis fangDhad favourable therapeutic action for OA, which mechanism of action possibly were relevant with promoting cartilage cell proliferation and recovery of cartilage cell DNA.Moreover,the osteoarthritis fang â–¡ had the ability of against hydrocortisone damage to cartilage cell, which provided theory foundation for its application uniting hydrocortisone.