Effect Of Eletroacupuncture With Close-to Bone Needling Treatment In Impaired Cartilage Of Rabbits With Knee Osteoarthritis

Objective:To investigate the effect of eletroacupuncture with close-to-bone needling treatment in impaired cartilage of rabbits with knee osteoarthritis(KOA)and explore its possible mechanisms.Methods:Forty New Zealand rabbits were randomized equally into Normal control group,KOA Modeling,eletroacupuncture with close-tobone needling group(CN group),and normal thrust needing group(NTN group).In the latter 3groups,KOA was induced by Hulth-Telhag treatmen t and evaluated with X-ray examination,and 6 weeks after the modeling,eletroacupuncture for 20 min was administered in CN and NTN groups at the acupoints "Zusanli" "waixiyan" "neixiyan" "liangqiu" and "yinlingquan" in the left knee joints once daily for 5days as a treatment cycle.after 4treatment cycles,the rabbits were examined for behavioral changes,carti lage morphology,and Mankin scores;The protein and mRNA expressions of Runx2,Sox9 and Col? were examined using Westen blotting,immu nohistochemistry,and RT-PCR as appropriate.Results:The analysis results of Lequesne MG score,HE staining,Mankin score,immunohistochemistry,WB and RT-PCR were basically consistent.(1)Lequesne MG shows that score of N group was 0~1 which was lowers that the score of M,CN,and NTN groups(P < 0.05).In comparison with the M groups,the Lequesne MG score had no obvious difference before the treatment and was significant lower in NTN,CN groups after the treatment(P < 0.05).(2)Staining observation:Articular cartilage surface of N group was smooth.Cartilage cells arranged in neat rows,and distributed evenly,tide line was complete and clear.In M group,The surface of cartilage cells were cracked,Cells arranged irregularly and Chondrocytes nested,Tide line blurred.CN and NTN groups cartilage cells arranged in neat rows,cells arranging in disorder still can be seen.Cartilage surface was smooth.Mankin score shows:The mankin score of articular cartilage from male patients in NM,CN and NTN group were 0~1,7~9,3~5 and 4~6.Four groups score were significantly different(P <0.01).(3)Immunohistochemistry and Western-Blot shows:Runx2 and Col?proteins expression in cartilage of M group significantly increased than that of N group,Two groups score were significantly different(P < 0.01).CN group decreased significantly compared with M group,and there were significant differences between the two groups(P < 0.01).There was significant difference between CN group and NTN group(P < 0.05).The results of immunohistochemistry and Western-Blot showed that eletroacupuncture with close-to-bone needling could effectively reduccd expression of Runx2 and Col?proteins,and the effect of CN group was better than NTN group.(4)RT-PCR experimental results showed: Sox9 mRNA expression in cartilage of M group significantly decreased than that of N group,Two groups score were significantly different(P < 0.01).CN group increased significantly compared with M group,and there were significant differences between the two groups(P < 0.01).There was significant difference between CN group and NTN group(P < 0.05).The results of RT-PCR showed that eletroacupuncture with close-to-bone needling method could effectively increaed expression of Sox9 mRNA,and the effect of CN group was better than NTN group.Conclusion:Eletroacupuncture with close-to-bone needling method could inhibit hypertrophic differentiation of chondrocyte through regulating the expression of Sox9 and Runx2,Col? of cartilage cell,which may be the possible therapeutic mechanism of KOA.Eletroacupuncture with close-to-bone needling method is more effective than normal thrust needing for KOA probably due to more intensive needling sensation.