College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China; Department of Microbiology and Plant Pathology, University of California-Riverside, Riverside, CA 92521, USA.
Phytomedicine. 2022 Sep;104:154296. doi: 10.1016/j.phymed.2022.154296. Epub 2022 Jul 2.
Apoptosis is thought to be involved in all processes, including normal cell cycle, immune system, atrophy, embryonic development, and chemical-induced cellular damage. However, if the normal apoptotic process fails, the results might be disastrous, e.g., chondrocytes damage in tibial dyschondroplasia (TD). TD is a worldwide issue in the poultry sector due to thiram toxicity. Thiram (Tetramethyl thiuram disulfide) is a dithiocarbamate pesticide and fungicide commonly used in horticulture to treat grains meant for seed protection and preservation.
According to prior studies, chlorogenic acid (CGA) is becoming essential for regulating apoptosis. But still, the specific role of CGA in chondrocyte cells remains unclear. The present study explored the molecular mechanism of CGA on chondrocytes' apoptosis with B-cell lymphoma 2 signaling under the effect of miR-460a.
An in vivo and in vitro study was performed according to our previously developed methodology. Flow cytometry, western blotting, reverse transcription-quantitative polymerase chain reaction, and immunofluorescence assay were used to investigate the involvement of apoptosis and inflammasome related pathways.
The CGA decreased the apoptosis rate with the deactivation of miR-460a, accompanied by the activation of Bcl-2. The high expression of miR-460a reduced the cell viability of chondrocytes in vitro and in vivo, that led to the interleukin-1β production. While the apoptotic executioners (caspase-3 and caspase-7) acted upstream in miR-460a overexpressing cells, and its depletion downgraded these executioners. The CGA administrated cells negatively regulated miR-460a expression and thus indicating the deactivation of the apoptotic and inflammasome related pathways.
Chlorogenic acid had a negative effect on miR-460a, setting off specific feedback to regulate apoptotic and inflammasome pathways, which might be a key feature for chondrocytes' survival.
细胞凋亡被认为参与了包括正常细胞周期、免疫系统、萎缩、胚胎发育和化学诱导的细胞损伤等在内的所有过程。然而，如果正常的细胞凋亡过程失败，结果可能是灾难性的，例如胫骨软骨发育不良（TD）中的软骨细胞损伤。TD 是由于 thiram 毒性而在禽类行业中存在的一个全球性问题。thiram（四甲基秋兰姆二硫化物）是一种二硫代氨基甲酸盐类杀虫剂和杀真菌剂，通常用于园艺业来处理用于种子保护和保存的谷物。
根据先前的研究，绿原酸（CGA）对于调节细胞凋亡变得至关重要。但是，CGA 在软骨细胞中的具体作用仍然不清楚。本研究探讨了 CGA 在 miR-460a 作用下通过 B 细胞淋巴瘤 2 信号对软骨细胞凋亡的分子机制。
根据我们之前开发的方法进行了体内和体外研究。流式细胞术、蛋白质印迹、逆转录定量聚合酶链反应和免疫荧光测定用于研究细胞凋亡和炎性小体相关途径的参与情况。
CGA 通过失活 miR-460a 降低了凋亡率，同时激活了 Bcl-2。miR-460a 的高表达降低了体外和体内软骨细胞的细胞活力，导致白细胞介素-1β的产生。而在 miR-460a 过表达细胞中，凋亡执行者（caspase-3 和 caspase-7）在上游起作用，而其耗竭则降低了这些执行者。给予 CGA 的细胞负调控 miR-460a 的表达，从而表明凋亡和炎性小体相关途径的失活。
绿原酸对 miR-460a 具有负效应，引发特定的反馈来调节凋亡和炎性小体途径，这可能是软骨细胞存活的关键特征。
