Song Xianbing, Zhang Bao, Taorong Wang, Wang Gaoyuan, Huang Yanping, Zhang Ye, Liu Meimei, Chen Xiaoyu
Department of Human Anatomy, Histology and Embryology, Anhui Medical College, China.
Department of Nutrition, the First Affiliated Hospital of Anhui Medical University, China.
Ann Clin Lab Sci. 2020 Jan;50(1):85-91.
The aim of the present study is to investigate the effect of cyclopamine, a hedgehog signaling pathway inhibitor, on adjuvant arthritis (AA), rat articular chondrocyte viability, and part mechanisms In this study, an AA rat model was established by Freund's complete adjuvant (FCA). The arthritis index (AI), secondary paw swelling degree, and HE staining were used to evaluate whether the model was successfully established. Chondrocytes of the ankle joint of AA rats were cultured and identified. Cyclopamine (0, 0.03, 0.1, 0.3, 1, 3, 10 and 30 mg/l) was administered to determine chondrocyte viability. Chondrocyte apoptosis was detected by Annexin V-FITC/PI double dye. The expression of hedgehog signaling pathway-related proteins Shh, Ptch1, and Gli1 in chondrocytes was detected by western blotting. The results show that AA was successfully induced by FCA since the AI of AA rats and secondary paw swelling degree increased and the cartilage tissue of the rats' ankle joint was damaged. Thus, the chondrocytes were successfully cultured following the identification of toluidine blue and type II collagen. Cyclopamine (0.03, 0.1, 0.3, 1, 3, 10 and 30 mg/l) could increase the viability of chondrocytes and reduce the apoptotic rate of chondrocytes. As compared with the control group, different doses of cyclopamine (0.3, 3 and 10 mg/l) significantly decreased the expression of Shh, Ptch1 and Gli1 proteins in AA chondrocytes. Therefore, an AA rat model was successfully established in the present study and cyclopamine improved the viability and inhibited the apoptosis of chondrocytes. This is an effect that may be associated with the inhibition of the chondrocyte hedgehog signaling pathway.
本研究旨在探讨刺猬信号通路抑制剂环杷明对佐剂性关节炎（AA）、大鼠关节软骨细胞活力的影响及其部分作用机制。在本研究中，通过弗氏完全佐剂（FCA）建立AA大鼠模型。采用关节炎指数（AI）、继发性爪肿胀程度及苏木精-伊红（HE）染色评估模型是否成功建立。对AA大鼠踝关节软骨细胞进行培养和鉴定。给予环杷明（0、0.03、0.1、0.3、1、3、10和30 mg/l）以测定软骨细胞活力。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）双染法检测软骨细胞凋亡情况。通过蛋白质免疫印迹法检测软骨细胞中刺猬信号通路相关蛋白音猬因子（Shh）、 patched 1（Ptch1）和glioma-associated oncogene 1（Gli1）的表达。结果显示，FCA成功诱导了AA，因为AA大鼠的AI和继发性爪肿胀程度增加，且大鼠踝关节软骨组织受损。因此，经甲苯胺蓝和II型胶原鉴定后，软骨细胞成功培养。环杷明（0.03、0.1、0.3、1、3、10和30 mg/l）可提高软骨细胞活力并降低软骨细胞凋亡率。与对照组相比，不同剂量的环杷明（0.3、3和10 mg/l）显著降低了AA软骨细胞中Shh、Ptch1和Gli1蛋白的表达。因此，本研究成功建立了AA大鼠模型，环杷明改善了软骨细胞活力并抑制了软骨细胞凋亡。这一作用可能与抑制软骨细胞刺猬信号通路有关。
