Mechanism Of Acupotomy Anti-Apoptosis Of KOA Rabbit Chondrocytes Based On P38/JNK MAPK Signal Pathway

Objective: To observe the effect of acupotomy on chondrocyte apoptosis and P38/JNK MAPK signal pathway in rabbits with knee osteoarthritis(KOA),and explore the mechanism of acupotomy on chondrocyte apoptosis in rabbits with KOA.Methods: 40 New Zealand white rabbits were randomly divided into normal group,model group,acupotomy group and sham acupotomy group,with 10 rabbits in each group.The KOA rabbit model was established using the Videman method.The acupotomy group selected the quadriceps femoris tendon,patellar ligament,and the strips and nodules around the internal and external collateral ligaments as the entry points.The sham acupotomy group avoided those in knee joint of the rabbit,and selected the entry points at the side.HE staining was used to detect the pathological changes of chondrocytes,and electron microscopy and TUNEL fluorescence staining were used to observe the apoptosis of chondrocytes.Real-time quantitative PCR(RT-PCR)and Western blot(WB)were used to detect the P38/JNK MAPK signal pathway and the mRNA and protein expression of P53,MMP-13,Bax,Bcl-2,caspase-3.Results: Compared with the normal group,the chondrocytes of KOA rabbits were arranged in disorder,the surface was uneven,and there were a large number of apoptotic chondrocytes marked by green fluorescence.There were obvious nuclear membrane shrinkage and chromatin edge collection in the chondrocytes.Compared with the model group,the pathological changes in the acupotomy group were alleviated,the number of green fluorescent labeled chondrocytes was reduced,and the chondrocytes were arranged in order,the nuclear membrane was smooth,and the chromatin was uniform.There was no significant difference in the sham acupotomy group.Western blot results showed that the protein expression of p-P38/P38,p-JNK/JNK,P53 and MMP13 in the model group was higher than that in the normal group(P<0.01);Compared with the model group,the protein expression of p-P38/P38,p-JNK/JNK,P53,and MMP13 in the acupotomy group was lower(P<0.01),while there was no difference in the protein expression of p-P38/P38,p-JNK/JNK,P53,and MMP13 in the sham acupotomy group(P>0.05).RT-PCR results showed that compared with the normal group,the mRNA expression of P38,JNK,P53,MMP13,Bax and caspase-3 in the model group was higher,while the mRNA expression of Bcl-2 was lower,P<0.01;Compared with the model group,the protein expression of P38,JNK,P53,MMP13,Bax and caspase-3 in the acupotomy group was lower,and the mRNA expression of Bcl-2 was higher,P<0.01;The mRNA expression of P38,JNK,P53,MMP13,MMP13,Bax,caspase-3,and Bcl-2 in the sham acupotomy group was not different from that in the model group(P>0.05);Compared with the acupotomy group,the mRNA expression of P38,JNK,P53,MMP13,MMP13,Bax and caspase-3 in the sham-acupotomy group was higher,while the mRNA expression of Bcl-2 was lower,P<0.01.Conclusion: Acupotomy therapy can alleviate the pathological changes of knee chondrocytes in rabbits with knee osteoarthritis and improve the degeneration of cartilage.Acupotomy therapy can significantly reduce the expression of pro-apoptotic protein Bax and caspase-3,increase the expression of anti-apoptotic protein Bcl-2,and play an antiapoptotic role in KOA rabbit chondrocytes by reducing the phosphorylation of P38/JNK MAPK signal pathway,down-regulating the expression of P53 and MMP-13.