Li Ping, Jiang Weiqian, Yang Qiming, Lu Yang, Zhang Jian
Department of Orthopaedics, The First Affiliated Hospital of Chongqing Medical University, No.1, You-Yi Road, district of Yu-Zhong, Chongqing, 400016, China.
Department of Orthopaedics, Chongqing Hospital of Traditional Chinese Medicine, Chongqing, 400021, China.
Heliyon. 2024 Aug 6;10(15):e35665. doi: 10.1016/j.heliyon.2024.e35665. eCollection 2024 Aug 15.
Leptin has been widely studied and found to have a significant impact on the development of osteoarthritis (OA). However, there are conflicting findings regarding the impact of leptin on chondrocytes. The study aimed to examine the impact of leptin on human chondrocytes and rats with OA. In the in vitro experiment, cartilage tissue obtained from patients hospitalized for knee replacement due to OA was collected for primary culture of chondrocytes. The proliferation and apoptosis of chondrocytes were assessed using cell counting kit-8 and flow cytometry. Autophagy levels were evaluated through monodansylcadaverine staining, mRFP-GFP-LC3 fluorescence, and transmission electron microscopy. Additionally, the expression of autophagy-related genes and proteins was analyzed using qRT-PCR and western blotting. In the in vivo experiment, an OA rat model was established. Following treatment with leptin and leptin antagonists, the cartilage tissues were examined using histology analysis (hematoxylin-eosin and Safranin O/fast green staining) and immunohistochemical. Mankin's score was utilized to assess the severity of OA, while qRT-PCR and western blotting were employed to detect the expression of autophagy-related genes and proteins in the cartilage. The ability of leptin to protect chondrocytes is achieved through the inhibition of autophagy via phosphoinositide 3 kinase/protein kinase B/mammalian target of rapamycin signaling pathway.
瘦素已得到广泛研究,并发现其对骨关节炎(OA)的发展有重大影响。然而,关于瘦素对软骨细胞的影响存在相互矛盾的研究结果。该研究旨在探讨瘦素对人软骨细胞和OA大鼠的影响。在体外实验中,收集因OA住院接受膝关节置换手术患者的软骨组织用于软骨细胞原代培养。使用细胞计数试剂盒-8和流式细胞术评估软骨细胞的增殖和凋亡。通过单丹磺酰尸胺染色、mRFP-GFP-LC3荧光和透射电子显微镜评估自噬水平。此外,使用qRT-PCR和蛋白质免疫印迹法分析自噬相关基因和蛋白质的表达。在体内实验中,建立OA大鼠模型。在用瘦素和瘦素拮抗剂治疗后,使用组织学分析(苏木精-伊红和番红O/固绿染色)和免疫组织化学检查软骨组织。采用Mankin评分评估OA的严重程度,同时使用qRT-PCR和蛋白质免疫印迹法检测软骨中自噬相关基因和蛋白质的表达。瘦素保护软骨细胞的能力是通过磷脂酰肌醇3激酶/蛋白激酶B/雷帕霉素哺乳动物靶标信号通路抑制自噬来实现的。