Objective To observe the effect of acupotomy on the expression of Beclin-1

Bcl-2 and Caspase-3 in the cartilage tissue in rabbits with knee osteoarthritis(KOA)

so as to explore its mechanism underling improvement of KOA. Methods Twenty-four healthy male New Zealand rabbits were randomly and equally divided into blank control

model and acupotomy groups

with 8 rabbits in each group. By using the modified Videman's methods

the KOA model was established by left hind limb immobilization with a plaster cast for 6 weeks. The severity of KOA(knee pain

swelling and motor function) was assessed using Lequesne score

and the rabbits with a score below 4 were excluded. The acupotomy was applied to “Hedingci”(the attachment of the quadriceps tendon to the patella at the upper edge)

“Binneixia”(the medial patellar supporting band attachment of medial inferior patellar margin)

“Binwaixia”(the lateral patellar supporting band attachment of the lower lateral patellar margin)

“Chengfeijian”(the lateral collateral ligament of the knee passes over the lateral joint space)

“Weiyangci”(the medial margin of biceps femoris at the lateral end of popliteus)

“Yinlingci”(the medial tibial attachment of anserinus tendon) on the left hind limb once a week for 4 weeks. One week after the last intervention

the left knee joint dysfunction severity(pain

maximum walking distance

and some activities of daily living) was evaluated by using modified Lequesne score. Histopathological changes of the cartilage were observed under light microscope after H.E. staining. The apoptosis of chondrocytes was observed after terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick-end labeling(TUNEL) staining. The autophagolysosomes of chondrocytes were observed using transmission electron microscopy. The expression levels of Beclin-1

Bcl-2 and Caspase-3(related factors of autophagy and apoptosis) were detected using Real-time PCR and Western blot separately. Results In comparison with the blank control group

the Lequesne score

apoptosis rate

expression levels of Caspase-3 mRNA and protein were significantly increased(P<0.001)

and the number of autophagolysosomes

expression levels of Beclin-1 and Bcl-2 mRNAs and proteins considerably decreased(P<0.001) in the model group. Relevant to the model group

the acupotomy group had an obvious decrease in Lequesne score

rate of apoptosis

and expression levels of Caspase-3 mRNA and protein(P<0.001) and an apparent increase in the number of autophagolysosomes and expression levels of Beclin-1 and Bcl-2 mRNAs and proteins(P<0.001). Findings of H.E. staining showed severe damaged cartilage surface

with a large number of exfoliation defects

few chondrocytes on the surface and disordered arrangement of transitional cells in the model group

which was relatively milder in the acupotomy group. Conclusion Acupotomy can mitigate knee-joint pain and improve functional activity in KOA rabbits

which may be associated with its functions in promoting autophagy and suppressing apoptosis by up-regulating expressions of Beclin-1 and Bcl-2 mRNAs and proteins and down-regulation of Caspase-3 mRNA and protein.