College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, PR China.
Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, 63100, Pakistan.
Phytomedicine. 2022 Jan;95:153865. doi: 10.1016/j.phymed.2021.153865. Epub 2021 Nov 23.
Tibial dyschondroplasia (TD) is a common disease characterized by proliferation and the deterioration of growth plate's chondrocytes due to widespread utilization of thiram in the agriculture and industrial sector.
In recent years, Nod-like receptor pyrin domain 3 (NLRP3) inflammasome has become a dilemma in the occurrence of many diseases. According to many research investigations, NLRP3 inflammasome has been linked to various diseases caused by pesticides and environmental toxins. Its involvement in such conditions opens up new treatment approaches. However, the role of the NLRP3 inflammasome in the development of TD is not fully understood under the impact of chlorogenic acid (CGA).
Chondrocytes were cultured with our previously developed methodology from growth plates. After morphological and molecular identification, chondrocytes were split into different groups to investigate the efficacy of chlorogenic acid. Cell apoptosis was determined through flow cytometry and Tunnel assay. Furthermore, RT-qPCR, immunofluorescence, and western blotting techniques were used to check marker genes and proteins expression.
In thiram-induced TD, Bax/Bak activation persuade a parallel pathway, mediated by the NLRP3 base inflammasome. It is worth mentioning that the apoptotic executioners (caspase-3 and caspase-7) act upstream for inflammasome. Furthermore, chondrocytes' ability to undergo mitochondrial apoptosis was governed by anti-apoptotic members, e.g., Bcl-2 and Bcl-xl. Equilibrium of these anti-apoptotic proteins ensured appropriate regulation of apoptosis during the development and survival of chondrocytes.
Chondrocytes have ability to undergo Bax/Bak-mediated apoptosis and generate pro-inflammatory signals, e.g., NLRP3 in thiram-induced TD. So, the Nod-like receptor pyrin domain 3 is the potential target to eliminate TD at all stages of pathology, while drugs, e.g., CGA, can significantly improve chondrocytes' survival by targeting these pro-inflammatory signals.
胫骨软骨发育不良（TD）是一种常见疾病，其特征是由于农业和工业部门广泛使用福美双，生长板的软骨细胞增殖和恶化。
近年来，Nod-like receptor pyrin domain 3（NLRP3）炎性小体在许多疾病的发生中成为一个难题。根据许多研究调查，NLRP3 炎性小体与农药和环境毒素引起的各种疾病有关。其在这些疾病中的作用为新的治疗方法开辟了道路。然而，在绿原酸（CGA）的影响下，NLRP3 炎性小体在 TD 发展中的作用尚不完全清楚。
采用我们之前开发的方法从生长板培养软骨细胞。在形态学和分子鉴定后，将软骨细胞分为不同组，以研究绿原酸的疗效。通过流式细胞术和Tunnel 检测法测定细胞凋亡。此外，还使用 RT-qPCR、免疫荧光和 Western blot 技术检查标记基因和蛋白的表达。
在福美双诱导的 TD 中，Bax/Bak 的激活促使 NLRP3 炎性小体介导的平行途径。值得注意的是，凋亡执行者（caspase-3 和 caspase-7）在炎性小体之前起作用。此外，抗凋亡成员（如 Bcl-2 和 Bcl-xl）控制着软骨细胞的线粒体凋亡能力。这些抗凋亡蛋白的平衡确保了在软骨细胞的发育和存活过程中适当调节凋亡。
软骨细胞具有 Bax/Bak 介导的凋亡能力，并产生促炎信号，例如福美双诱导的 TD 中的 NLRP3。因此，NLRP3 是在病理的所有阶段消除 TD 的潜在靶标，而药物，如 CGA，通过靶向这些促炎信号，可显著改善软骨细胞的存活。
