Effect of mechanical pressure on expression of signal transduction molecule Casaase-3, bcl-2, bax mRNA
Xiong-feng Li, Rujie Zhuang, Wei Zhuang
Abstract
Xiong-feng Li, Rujie Zhuang, Wei Zhuang
Abstract
Objective To explore the effects of mechanical stimuli on the expression of signal transduction molecule Caspase-3,B-cell lymphoma-2 ( bcl-2),bax mRNA.Methods Rabbit knee articular cartilage cells obtained by enzymatic digestion were cultured and passaged for 3 generations.The chondrocytes were cultivated in a culture plate added with different doses of inhibitor of Caspase-3,bcl-2,bax by a closed container with 90 kPa pressure for 24 h.Chondrocytes apoptosis was detected by flow cytometry (FCM),and the expression of Caspase-3,bcl-2,bax mRNA by semiquantitative analysis of polymerase chain reaction (PCR).Results Mechanical pressure could induce chondrocytes apoptosis.There was significant difference in apoptosis rate between inhibitors-treated groups and contral group ( all P < 0.05 ).There was significant difference in the expression of signal transduction molecule Caspase-3,bcl-2 and bax between inhibitors-treated groups and control group ( all P < 0.05).There were correlations between apoptosis rate and expression of Caspase-3,bcl-2 and bax in Caspase-3,bcl-2 and bax inhibitors-treated groups respectively ( for Caspase-3,r =0.69,t =3.41,P < 0.01 ; for bcl-2,r =0.73,t =3.97,P < 0.O1 ; for bax,r =0.89,t =6.69,P<0.01).Conclusion The inhibitors of Caspase-3,bcl-2 and bax can influence the apoptosis of chondrocytes,and make the expression of Caspase-3,bcl-2 and bax mRNA change. Key words: Chondrocyte; Mechanical pressure; Apoptosis
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Objective To explore the effects of mechanical stimuli on the expression of signal transduction molecule Caspase-3,B-cell lymphoma-2 ( bcl-2),bax mRNA.Methods Rabbit knee articular cartilage cells obtained by enzymatic digestion were cultured and passaged for 3 generations.The chondrocytes were cultivated in a culture plate added with different doses of inhibitor of Caspase-3,bcl-2,bax by a closed container with 90 kPa pressure for 24 h.Chondrocytes apoptosis was detected by flow cytometry (FCM),and the expression of Caspase-3,bcl-2,bax mRNA by semiquantitative analysis of polymerase chain reaction (PCR).Results Mechanical pressure could induce chondrocytes apoptosis.There was significant difference in apoptosis rate between inhibitors-treated groups and contral group ( all P < 0.05 ).There was significant difference in the expression of signal transduction molecule Caspase-3,bcl-2 and bax between inhibitors-treated groups and control group ( all P < 0.05).There were correlations between apoptosis rate and expression of Caspase-3,bcl-2 and bax in Caspase-3,bcl-2 and bax inhibitors-treated groups respectively ( for Caspase-3,r =0.69,t =3.41,P < 0.01 ; for bcl-2,r =0.73,t =3.97,P < 0.O1 ; for bax,r =0.89,t =6.69,P<0.01).Conclusion The inhibitors of Caspase-3,bcl-2 and bax can influence the apoptosis of chondrocytes,and make the expression of Caspase-3,bcl-2 and bax mRNA change. Key words: Chondrocyte; Mechanical pressure; Apoptosis
Key concepts: Apoptosis, Chondrocyte, Molecular biology, Messenger RNA, Caspase 3, Flow cytometry, Chemistry, Caspase-9