Research Bushengyiqihuoxue Law And Human Chondrocyte Proliferation MAPKs Signal Pathway For The Intervention Of Osteoarthritis

Osteoarthritis (Osteoarthritis,OA)is a chronic progressive degenerative disease. The main show is joint pain, stiffness, swelling, and restricted movement. Traditional chinese medicine holds that osteoarthritis always belongs to the category of "bi disease". Traditional Chinese medicine treatment have advantages of curative effect and less side effects. It has great potential in alleviating clinical symptoms and delaying the articular cartilage injury of OA. My teacher Haibo Yin has rich clinical experience in the treatment of OA. Tonifying Kidney, Tonifying Qi and Invigorate the circulation of blood are his main treatmen experience for OA.We choose YINGYANGHUO、HANGQI、DANSHEN as representative of Tonifying Kidney、Tonifying Qi and Invigorate the circulation of blood. This research established OA chondrocyte system in vitro as the object,and observed the effect of Tonifying Kidney、Tonifying Qi and Invigorate the circulation of blood on cartilage cell proliferation and Mitogen-activated protein kinase signaling pathways.Experimental study:Objective:To establish a finite cell line of human osteoarthritis chondrocytes, and observe the growth and morphology characteristic of chondrocytes; Discussing the molecular biology mechanism of icariin、astragaloside A-. tanshinone IIA on how to slow down the cartilage damage.Methods:Cartilage cell culture; Detect the proliferation of Cartilage cell by CCK-8; Detect the protein expression of MAPKs signaling pathway in Cartilage cell by Western Blotting.Results:1Successfully established a finite cell line of human osteoarthritis chondrocytes, which growth in good condition and had stable biological property. Compared with normal cartilage cells, the growth rate of OA cartilage cells is significantly slower. OA cartilage cells has high expression of p38、p-p38、 JNK、P-JNK、ERK、P-ERK, and phosphorylation of p38、ERK and JNK was significantly higher than that of in normal cartilage cells.2The proliferation of cartilage cells have been affected by icariin. In24h-96h, concentration of10to20ng/mL icariin can induce the proliferation and the concentration of40-80ng/mL of icariin can inhibit the proliferation. 10ng/ml、20ng/ml、40ng/ml、80ng/ml icariin can inhibit the expression of p38、 p-p38and the phosphorylation of p38. lOng/ml、80ng/ml icariin can inhibit the expression of JNK,20ng/ml、40ng/ml icariin can induce the expression of JNK.10-80ng/mL can inhibit the expression of pJNK.10ng/mL、20ng/mL、40ng/mL icariin can inhibit the phosphory lat ion of JNK.10ng/ml、20ng/ml、40ng/ml、80ng/ml icariin can inhibit the expression of ERK、 p-ERK and the phosphorylation of ERK.3The proliferation of cartilage cells have been affected by astragaloside A. In24h-48h, concentration of10to80ng/mL astragaloside A can induce the proliferation, In48h-96h, cartilage cell to apoptosis,10to80ng/mL astragaloside A can inhibit the apoptosis.10ng/ml、20ng/ml、40ng/ml、80ng/ml astragaloside A can inhibit the expression of p38、p-p38and the phosphory lat ion of p38.10ng/ml、20ng/ml、40ng/ml、80ng/ml astragaloside A can inhibit the expression of JNK. p-JNK.10to40ng/mL astragaloside A can inhibit the phosphorylation of JNK.10ng/ml、20ng/ml、40ng/ml、80ng/ml astragaloside A can inhibit the expression of ERK. p-ERK and the phosphorylation of ERK.4The proliferation of cartilage cells have been affected by tanshinone IIA. In24h-96h, concentration of10to20ng/mL tanshinone IIA can induce the proliferation and the concentration of40-80ng/mL of tanshinone IIA can inhibit the proliferation. lOng/ml,20ng/ml icariin can induce the expression of p38, p-p38,20ng/ml、80ng/ml tanshinone IIA can inhibit the expression of p38、p-p38,10ng/mL、40ng/mL、80ng/mL tanshinone IIA can inhibit the phosphorylation of p38.10-80ng/ml tanshinone IIA can not inhibit the expression of JNK、p-JNK and the phosphorylation of JNK.10ng/ml、20ng/ml、40ng/ml、80ng/ml tanshinone IIA can inhibit the expression of ERK、p-ERK and the phosphorylation of ERK.Conclusion:Compared with normal cartilage cells, the proliferation rate of OA cartilage cells is significantly slower. The activation of MAPKs signaling pathways in OA cartilage cell is hingher than normal cartilage cells. The low proliferation of cartilage cells and the high activation of MAPKs signaling pathways is the molecular mechanism of destruction of articular cartilage.Icariin、astragaloside A、 tanshinone IIA can delay the damagement of cartilage by controlling the rate of cartilage cell apoptosis and the activation of MAPKs signaling pathways in cartilage cell.