Muhammad Fakhar-E-Alam Kulyar1, Quan Mo1, Wangyuan Yao2, Yanmei Ding1, Zhang Yan1, Haitao Du1, Huachun Pan1, Kewei Li1, Jindong Gao1, Muhammad Shahzad3, Muhammad Khalid Mansoor3, Mudassar Iqbal4, Muhammad Waqas5, Muhammad Akhtar1, Zeeshan Ahmad Bhutta6, Jiakui Li7. 1. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China. 2. 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. 3. Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, 63100, Pakistan. 4. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China; Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, 63100, Pakistan. 5. Faculty of Veterinary & Animal Sciences, University of Poonch Rawalakot, Azad Jammu & Kashmir, 12350, Pakistan. 6. College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea. 7. 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. Electronic address: lijk210@sina.com.
Abstract
BACKGROUND: 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. PURPOSE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
BACKGROUND: 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. PURPOSE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.