Zhou Jingming, Chen Qian, Lanske Beate, Fleming Braden C, Terek Richard, Wei Xiaochun, Zhang Ge, Wang Shaowei, Li Kai, Wei Lei
Arthritis Res Ther. 2014 Jan 15;16(1):R11. doi: 10.1186/ar4437.
Previous observations implicate Indian hedgehog (Ihh) signaling in osteoarthritis (OA) development because it regulates chondrocyte hypertrophy and matrix metallopeptidase 13 (MMP-13) expression. However, there is no direct genetic evidence for the role of Ihh in OA, because mice with cartilage or other tissue-specific deletion of the Ihh gene die shortly after birth. We evaluated the role of Ihh in vivo via a Cre-loxP-mediated approach to circumvent the early death caused by Ihh deficiency.
To evaluate the role of Ihh in OA development, Ihh was specifically deleted in murine cartilage using an Ihh conditional deletion construct (Col2a1-CreER(T2); Ihh(fl/fl)). The extent of cartilage degradation and OA progression after Ihh deletion was assessed by histological analysis, immunohistochemistry, real-time PCR and in vivo fluorescence molecular tomography (FMT) 2 months after OA was induced by partial medial meniscectomy. The effect of Ihh signaling on cartilage was compared between Ihh-deleted mice and their control littermates.
Only mild OA changes were observed in Ihh-deleted mice, while control mice displayed significantly more cartilage damage. Typical OA markers such as type X collagen and MMP-13 were decreased in Ihh-deleted mice. In vivo FMT demonstrated decreased cathepsins and MMP activity in knee joints of animals with deletion of Ihh.
These findings support the protective role of Ihh deletion in surgically induced OA. Thus, our findings suggest the potential to develop new therapeutic strategies that can prevent and treat OA by inhibiting Ihh signaling in chondrocytes.
先前的观察表明,印度刺猬因子(Ihh)信号传导参与骨关节炎(OA)的发展,因为它调节软骨细胞肥大和基质金属蛋白酶13(MMP - 13)的表达。然而,尚无Ihh在OA中作用的直接遗传学证据,因为Ihh基因在软骨或其他组织特异性缺失的小鼠出生后不久即死亡。我们通过Cre - loxP介导的方法评估Ihh在体内的作用,以规避Ihh缺乏导致的早期死亡。
为了评估Ihh在OA发展中的作用,使用Ihh条件性缺失构建体(Col2a1 - CreER(T2); Ihh(fl/fl))在小鼠软骨中特异性删除Ihh。在通过部分内侧半月板切除术诱导OA 2个月后,通过组织学分析、免疫组织化学、实时PCR和体内荧光分子断层扫描(FMT)评估Ihh缺失后软骨降解程度和OA进展情况。比较Ihh缺失小鼠与其对照同窝仔鼠中Ihh信号传导对软骨的影响。
在Ihh缺失的小鼠中仅观察到轻度OA变化,而对照小鼠显示出明显更多的软骨损伤。Ihh缺失小鼠中典型的OA标志物如X型胶原蛋白和MMP - 13减少。体内FMT显示Ihh缺失动物膝关节中的组织蛋白酶和MMP活性降低。
这些发现支持Ihh缺失在手术诱导的OA中的保护作用。因此,我们的发现表明有可能开发新的治疗策略,通过抑制软骨细胞中的Ihh信号传导来预防和治疗OA。