Effects Of Semen Strychni On Apoptosis And Proliferation Of Chondrocyte In Osteoarthritis

Objective:The effect of Tongluo drug Semen strychni including its major monomer Brucine,Strchnine on apoptosis and synthesis metabolic function of chondrocyte was investingated in osteoarthritis.Methods:1.The protect effect of Semen Strychni on knee OA in rabbit was observed in vivi.The rabbit osteoarthritis model were duplicated by Hulth's technique .The pathologyical changes of each group was observed with naked eyes before taking the thighbone department carilage of pulley block and the platform place of shine bone carilage,then under the light microcope and the transmission electron microcope respectively,after treating with Semer Strychni(high,middle and low dose)for two months.2.The effect of Tongluo drug Semen Strychni on apoptosis and synthesis metablic function of chondrocyte was investingated in osteoarthritis.Firstly,the apoptosis and proliferation of normal chondrocytes were observed by flow cytometry and MTT respectively after treated with Semen strychni ,Brucine and Strchnine.Secondly,the three chondrocyte apoptosis models were established by using different drugs,treated with sodium nitroprusside(SNP) ,all-trans retinoic acid (ATRA)and tumor necrosis factor－ a (TNF－a) respectively. The effects of Semen strychni ,Brucine and Strchnine on chondrocytes apoptosis induced by different ways were observed.The apoptosis rate were analysed by AnnexinⅤ/PI with flaw cytometry and the characteristic alterations of morphology were detected by electron miscroscope and DNA agarose gel electrophoresis respectively, and through assessing NO content of three kinds of apoptosis models. The relation between the apoptosis models and Semen strychni further action mechanicsm on chondrocyte apoptosis were known.Finally,DNA content,collage and proteoglycan of chondrocyte were detected by 3H-Tdr,3H－proline,35S－sulfate isotope incorporation.The synthetic metabolic change of chondrocytes were observed in course of its apoptosiss and synthetic metabolic function of chondrocytes were also observed when Semen strychni regulating the apoptosis and proliferation of chondrocyte.Results:1.It was found that Semen strychni had protecting action on knee OA in rabbit,the high and middle dosage of Semen strychni could relieve effect on artilage degeneration2.The high,middle,low dosage of Semen strychni ,Brucine and Strchnine could had no effects on apoptosis of normal chondrocyte in vitro. Strchnine could induce to death for normal chondrocyte.3. Semen strychni and Brucine could stimulate cell proliferation to some extent, Brucine had dose-dependent on increase of choncytes,and high dosage of Brucine was apparently, Strchnine could inhabit cell proliferateion,and high dosage of Strchnine was apparently.4.When rabbit chondrocyte were incubated with SNP(2mmol/l),and ATRA(10-4mol/l),and chondrocyte coming from fetal were incubated with TNF-α(30ng／ml) for 24h,they all could induce apoptosis in chondrocyte in vitro. Semen strychni and Brucine could decreas early apoptosis induced by SNP,ATRA at high and middle dosage,and high dosage was better,Strchnine couldn't inhabit apoptosiss induced by SNP and ATRA.Semen strychni ,Brucine and Strchnine all had no effects on apoptosis induced by TNF-α.5.It was found that chondrocyte apoptosis was induced by SNP,TNF－αthrogh NO way,relating with NO going up obviously,after detecting the content of NO of three models of apoptosis chondrocyte.The apoptosis was induced byATRA not through NO,and not relying on NO. Semen strychni and Brucine could inhabit apoptosis induced by different ways.6. The high and middle dosage of Semen strychni , Brucine could promote DNA and proteoglycan synthesis of chondrocyte apparently.they had dose-dependent,and the collagen synthesis of chondrocyte was inhabited bySemen strychni,Brucine.Chondrocyte apoptosis was occurring whereas the capacity of chondrocyte proliferation and synthesis decreased. Semen strychni , Brucine could improve synthetic metabolic function when they were inhabiting apoptos...