The Role Of TRPM4 Channel Inhibitor 9-Phenanthrenol In OA Chondrocytes

Objective:1、To reseach whether(transient receptor potential M4)TRPM4 signal channel inhibitor-9-phenanthrol can cartilage cell hypertrophy and matrix degeneration of cartilage explants in human osteoarthritis(OA).2、To reseach the effect of 9-phenanthrol on apoptosis of OA chondrocytes,so as to provide experimental basis for treatment of OA;Methods:1、The human OA chondrocytes derive from subjects with surgery of total knee replacement.2、Culture the cells in the presence of blank group and other five group which contain different concentrations of 9-phenanthrol such as 5×* 10-6mol/l、l* 10-5 mol/l、2* 10-5mol/l、4* 10-5 mol/land8×10-5 mol/l.3、MMP-13,Runx-2,Aggrecan,COL II mRNA were determined by real-time qPCR after 24 h of incubation.4 、The effects of 9-phenanthrol on the apoptosis of OA chondrocytes were detected by TUNEL staining and fluorescence microscopy and Annexin V and 7-AAD labeled flow cytometry after 1 days of incubation with 9-phenanthrol;Results:1、The RT-qPCR results showed the effect of 9-phenanthrol on hypertrophic differentiation and cell metabolism on chondrocyte OA by inhibiting TRPM4 ion channel: 9-phenanthrol group cartilage in 8* 10-5 mol/l concentration in AGG cells compared with blank control group,the expression level was significantly lower(P<0.05)and other groups with different concentration was also declined.At the same time,the concentration of COLII in the 9-group at the concentration of 8 * 10-5 mol/l was significantly higher(P<0.05)and other groups had an upward trend than that in the control group except in the 10-6mol/l group.so it is unable to judge the impact of 9-phenanthrol on chondrocyte metabolism;and chondrocyte hypertrophy index Runx-2,MMP13,mRNA compared with the control group the expression level increased visible,9-phenanthrol may promote chondrocyte hypertrophy at the mRNA level.2、Effect of 9-phenanthrol on apoptosis of OA chondrocytes: TUNEL staining of cartilage cell apoptosis rate of fluorescence microscopy showed that compared with the control group the apoptosis rate of the 9-phenanthrol group at concentration of 2 * 10-5mol/l was significantly decreased(P<0.05),other groups compared with the control group had no statistical significance,but has the apoptotic rate decreased.The rate of detection of cartilage cell apoptosis cytometry Annexin V and 7-AAD 9-labeled flow,compared with the control group,the apoptosis rate of the 9-phenanthrol group at concentration of l* 10-5 mol/l、2* 10-5mol/l、4* 10-5 mol/l decreased(P<0.05),and decreased in the other two groups.Conclusion:1 、 9-Phenanthrenol promotes the proliferation and differentiation of OA chondrocytes.2、9-can reduce the apoptosis of phenanthrol OA chondrocytes.