我们发现抗代谢增殖药羟喜树碱可通过诱导成纤维细胞凋亡、防治膝关节术后瘢痕粘连。采用基因芯片技术检测羟喜树碱刺激人成纤维细胞后的miRNA表达谱时发现miR-23b低表达于正常成纤维细胞。前期研究发现抑制miR-23b功能可以增加药物诱导成纤维细胞的凋亡率，生物信息学分析及荧光素酶报告基因系统验证结果发现并证实SPRY2是miR-23b的靶基因。SPRY2是Ras-MAPK信号通道的重要抑制因子，可诱导细胞凋亡。我们推测miR-23b可能通过调控SPRY2而减少膝关节粘连。本研究拟探讨miR-23b在人成纤维细胞中生物学特征；在体观察miR-23b与膝关节内瘢痕粘连的关系；并采用基因过表达及敲除技术研究miR-23b通过作用于SPRY2介导成纤维细胞凋亡的分子机制。miR-23b影响膝关节粘连及其机制的研究未见报道，本研究可为关节瘢痕粘连及其并发症的防治提供新思路及潜在治疗靶点。

We found that Hydroxycamptothecin, an anti-metabolic and anti-proliferated drug, could prevent intraarticular scar adhesion by inducing fibroblast apoptosis after knee surgery. The microarray results showed that miR-23b expression in human fibroblasts stimulated by hydroxycamptothecin was lower than that in normal fibroblasts. Our previous study showed that inhibition of miR-23b expression could increase apoptosis of fibroblast. SPRY2 was found and confirmed that it was a target gene of miR-23b by bioinformatics analysis and luciferase reporter gene system. SPRY2 is an important inhibitor of the Ras-MAPK signal pathway and can induce cell apoptosis. We speculate that miR-23b can prevent intraarticular scar adhesion by regulating SPRY2. The study was to investigate the biological characteristics of miR-23b in human fibroblasts, identify the relationship between miR-23b and intraarticular scar adhesion in animal model and investigate the mechanism of miR-23b inducing fibroblast apoptosis by regulate SPRY2 expression through gene overexpression and knockout technology. There was no report about the mechanism of miR-23b on intraarticular adhesion.The study could provied an new and potential therapeutic target for scar adhesion and its complications.

研究背景及目的：成纤维细胞过度增殖是膝关节术后瘢痕粘连形成的主要原因。抗代谢增殖药物羟基喜树碱（HCPT）能够通过诱导成纤维细胞凋亡防治膝关节术后瘢痕粘连。本研究探讨miR-23b在成纤维细胞中的生物学特征及其影响膝关节瘢痕粘连的机制。实验方法：CCK-8检测HCPT对人成纤维细胞活性的影响。Western blot、Hoechst 33342染色检测HCPT诱导成纤维细胞凋亡的情况。qRT-PCR检测miR-23b表达。Western blot、流式细胞技术检测过表达和沉默miR-23b对成纤维细胞凋亡的影响及其对相关信号通道的调控作用，并分析miR-23b在HCPT诱导成纤维细胞凋亡中的作用。Western blot方法检测miR-23b对Smad3的调控表达。组织学观察、TUNEL染色、羟脯氨酸含量测定、qRT-PCR检测评价HCPT调控miR-23b减少兔膝关节瘢痕粘连的情况。实验结果：HCPT显著降低人成纤维细胞活性。Western blot、Hoechst 33342染色和流式细胞技术结果显示HCPT能够诱导人成纤维细胞凋亡。qRT-PCR结果显示HCPT能够上调miR-23b表达；TGF-β1能够下调miR-23b表达。Western blot、流式细胞技术显示上调miR-23b表达和HCPT均能诱导成纤维细胞凋亡，抑制miR-23b能够减少成纤维细胞凋亡，并能降低HCPT诱导的成纤维细胞凋亡。miR-23b能够下调Smad3表达，Smad3是miR-23b的靶基因。HCPT能够上调miR-23b表达，从而抑制成纤维生长、减少兔膝关节术后瘢痕粘连。结论：成纤维细胞增殖状态时miR-23b表达下调，成纤维细胞凋亡状态时miR-23b表达上调；miR-23b过表达能够诱导成纤维细胞凋亡，而抑制miR-23b表达能够减少成纤维细胞凋亡，并能降低HCPT诱导的成纤维细胞凋亡；HCPT能够上调miR-23b表达，从而抑制成纤维细胞生长、减少兔膝关节术后瘢痕粘连。本研究结果可为瘢痕粘连的防治提供新思路和潜在治疗靶点。