Department of Orthopedics Surgery, Shaanxi Provincial People's Hospital, Xi'an JiaoTong University, Xi'an, People's Republic of China.
Institute of Endemic Diseases of School of Public Health, Health Science Center of Xi'an JiaoTong University, NHC Key Laboratory of Trace Elements and Endemic Diseases, Xi'an, Shaanxi, People's Republic of China.
Cartilage. 2021 Dec;13(1_suppl):789S-796S. doi: 10.1177/1947603519886626. Epub 2019 Nov 24.
. To explore the relationship between the death receptor (DR) and the pathological progression of Kashin-Beck disease (KBD). . KBD cartilage samples were collected from 15 patients diagnosed according to the "National Diagnostic Criteria of KBD" in China. monolayer chondrocytes were cultured in complete medium. Caspase-3 and caspase-8 activities in chondrocytes were analyzed using a kit. Nuclear morphology was observed by Hoechst 33258 staining, apoptosis was verified by flow cytometry analysis, and DR molecules were detected using Western blotting and quantitative real-time reverse transcription polymerase chain reaction analysis. . Early apoptotic rates of KBD and osteoarthritis (OA) chondrocytes were higher than those of normal control (NC) cells. Excessive apoptotic nuclei were observed in OA and KBD cells after Hoechst 33258 staining. Activities of both caspase-3 and caspase-8 were higher in KBD and OA cells than in NC cells. The average DR4 mRNA level in KBD cells was 3.301-fold higher than that in NC cells, Fas-associating protein with death domain (FADD) transcript level in KBD cells was 2.528-fold higher than that in NC cells. Western blot analyses showed that FAS, DR4, DR5, caspase-3, and FADD were upregulated in the KBD and OA groups compared with the NC group. High expression of caspase-8 in KBD compared with NC was verified, whereas cellular FLICE-inhibitory protein (c-FLIP) in KBD was significantly downregulated. . KBD and OA chondrocytes showed obvious FADD-caspase-dependent apoptosis, which is related to the DR pathway. Apoptosis in KBD articular cartilage is mainly related to FAS/DR4-FADD-caspase signaling, and OA is associated with FAS/DR4/DR5-FADD-caspase signaling.
探讨死亡受体（DR）与大骨节病（KBD）病理进展的关系。
从中国“大骨节病诊断国家标准”诊断的 15 例患者中采集 KBD 软骨样本。使用完全培养基培养单层软骨细胞。使用试剂盒分析软骨细胞中 caspase-3 和 caspase-8 的活性。通过 Hoechst 33258 染色观察核形态，通过流式细胞术分析验证凋亡，通过 Western blot 和实时定量逆转录聚合酶链反应分析检测 DR 分子。
KBD 和骨关节炎（OA）软骨细胞的早期凋亡率高于正常对照（NC）细胞。Hoechst 33258 染色后，OA 和 KBD 细胞中观察到过多凋亡核。KBD 和 OA 细胞中 caspase-3 和 caspase-8 的活性均高于 NC 细胞。KBD 细胞中平均 DR4mRNA 水平比 NC 细胞高 3.301 倍，KBD 细胞中 Fas 相关死亡结构域蛋白（FADD）转录水平比 NC 细胞高 2.528 倍。Western blot 分析显示，与 NC 组相比，KBD 和 OA 组的 FAS、DR4、DR5、caspase-3 和 FADD 表达上调。与 NC 相比，KBD 中 caspase-8 的高表达得到验证，而 KBD 中的细胞 FLICE 抑制蛋白（c-FLIP）明显下调。
KBD 和 OA 软骨细胞表现出明显的 FADD-caspase 依赖性凋亡，这与 DR 途径有关。KBD 关节软骨中的凋亡主要与 FAS/DR4-FADD-caspase 信号有关，OA 与 FAS/DR4/DR5-FADD-caspase 信号有关。
