研究TRPM4信号通道抑制剂-9-菲酚对骨关节炎（OA）软骨细胞的肥大分化及软骨组织的降解的作用，并研究9-菲酚对OA软骨细胞凋亡的影响，从而为治疗OA提供实验依据。

选取因OA进行膝关节置换而截下的胫骨平台关节软骨，消化成软骨细胞进行细胞培养。软骨细胞均分为6组，一组不加药作为对照组，其余分别加入不同量的。9-菲酚干预1 d培养后细胞分别进行RT-qPCR，检测Runx-2、MMP-13（软骨细胞肥大指标）及COL II、Aggrecan（软骨代谢指标）；采用TUNEL染色荧光显微镜检测及Annexin V和7-AAD标记流式细胞仪检测9-菲酚干预1 d后OA软骨细胞凋亡情况。

RT-qPCR显示：无法判断9-菲酚对软骨细胞代谢的影响；而软骨细胞肥大指标Runx-2、MMP13的mRNA较对照组表达水平有升高趋势，可见在mRNA水平上，9-菲酚可能促进软骨细胞肥大。TUNEL染色荧光显微镜检测软骨细胞凋亡率发现，9-菲酚浓度为2×10^-5 mol/L组较对照组凋亡率明显下降（P<0.05），其他浓度组与对照组比较均无统计学差异，但均有凋亡率下降的趋势。Annexin V和7-AAD标记流式细胞仪检测软骨细胞凋亡率发现，9-菲酚浓度为1×10^-5 mol/L、2×10^-5 mol/L、4×10^-5 mol/L时较对照组凋亡率均有所下降（P<0.05），而在另外两组有下降趋势。

9-菲酚在细胞水平上对OA软骨细胞的肥大分化有促进作用，可能促进OA的进展。同时9-菲酚可以抑制OA软骨细胞的凋亡，在OA治疗方面可能具有积极意义。

To investigate whether TRPM4 channel inhibitor 9-phenanthrol can attenuate human osteoarthritis (OA) chondrocytes hypertrophy and cartilage explant matrix degeneration and to study the effect of 9-phenanthrol on apoptosis of OA chondrocytes to provide an experimental basis for the treatment of OA.

Human OA chondrocytes and cartilage explants, obtained from subjects with OA undergoing total knee arthroplasty, were cultured in the absence (blank group) or presence of different concentrations of 9-phenanthrol. Runx-2, MMP-13, COL II, and aggrecan mRNA expression was determined by real-time quantitative PCR (RT-qPCR) after 24 h of incubation. The effect of 9-phenanthrol on the apoptosis of OA chondrocytes was detected by TUNEL staining with fluorescence microscopy and flow cytometry with Annexin V and 7-AAD double staining after 1 day of incubation with 9-phenanthrol.

According to RT-qPCR results, it was unable to judge the effect of 9-phenanthrol on hypertrophic differentiation and cell metabolism in OA chondrocytes, and the mRNA expression levels of chondrocyte hypertrophy indexes Runx-2 and MMP13 increased compared with the control group, suggesting that 9-phenanthrol may promote chondrocyte hypertrophy at the mRNA level. TUNEL staining with fluorescence microscopy showed that compared with the control group, the apoptosis rate of cells treated with 2×10^-5 mol/L 9-phenanthrol was significantly decreased (P < 0.05), while no significant difference was observed in other concentration groups, although there was a decreasing trend compared with the control group. Flow cytometry with Annexin V and 7-AAD double staining showed that compared with the control group, the rate of apoptotic cartilage cells in the l×10^-5 mol/L, 2×10^-5 mol/L, and 4×10^-5 mol/L 9-phenanthrol groups significantly decreased (P < 0.05), while no significant difference was observed in other two concentration groups, although there was a decreasing trend compared with the control group.

9-phenanthrenol promotes the proliferation and differentiation of OA chondrocytes at the cellular level and may promote the development of OA. However, 9-phenanthrenol can also reduce the apoptosis of OA chondrocytes and thus has positive significance in the treatment of OA.