The Study On The Expression Of Protein Kinase B And Protein Kinase C In Human Osteoarthritis Cartilage

Objective With detecting the expression and relationship of PKB/Akt and PKC in knee osteoarthritis(OA), We investigating the role of PKB/Akt and PKC in the pathogenesis of OA, in order to provide some evidence for clinical therapy at the molecular level.Methods 38 samples of OA human cartilage were collected from total knee arthroplasty as experimental group. 11 samples of normal human cartilage were collected from patients were amputated because of unintentional injuries as control group. All samples were selected from the condyle of femur. The normal HE and safranin-o staining were used to observe the histological patterns of articular cartilage; the distribution of PKB/Akt and PKC in the chondrocyte were observed under Laser scanning confocal microscope. TUNEL metheod were used to detect the chondrocyte apoptosis; S-P immunohistochemistry staining and WesternBlotting were used to detect the expression of PKB/Akt and PKC protein in cartilage. Image-Pro Plus 6.0 and Gel pro 4.5 were used to make quantitative analysis. Independent Two-sample t-test,Paired Two-sample t-test and Pearson correlation tests were used for statics.Results①The histological change: the surface of cartilage is smooth and flat in normal articular,its chondrocyte were uniformly distributed with full tide line and matrix Integrity;conversely,The surface of cartilage is rough even absence in OA articular,chondrocyte distributes abnormal,accompany with fracture tidy mark and matrix losing.②The apoptosis of chondrocytes in cartilage: the percentage of apoptosis in OA cartilage is higher than normal cartilag(et=6.551,P<0.05). no matter in OA or normal human cartilage, the percentage of apoptosis in radical zone is higher than transitional zone(t=5.671,t=5.237, p<0.05).③The expression of PKB/Akt and PKCαprotein: the expression value of PKB/Akt in normal human cartilage is significant higher than in OA human cartilage(t=9.848,P<0.05),However, the phosphorylation of PKB/Akt increases in OA human cartilage;the expression value of PKCαin normal human cartilage is significant higher than in OA human cartilage (t=6.401,P<0.05).④The distribution of PKB/Akt and PKCαprotein: PKB/Akt and PKCαare mainly distributed in the middle zone and deep zone in normal human and OA cartilage. No matter in normal or OA cartilage, The expression of PKB/Akt in transitional zone is significant higher than radical zone(t=44.296,P<0.05) (t=5.150,P<0.05); The expression of PKCαin middle zone is significant higher than deep zone(t=5.343,P<0.05) (t=9.289,P<0.05).⑤The distribution of Akt and PKCαis mainly in cytoplasma.Conclusion The expression of PKB/Akt protein in OA human cartilage was significantly lower. Nevertheless, the expression of PKCαin OA human cartilage was significantly higher. These data indicated that Akt and PKCαinvolved in the regulation of chondrocyte apoptosis in human OA process.They may be one of the therapeutic potential targets for human OA.