Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid
Abstract Background The B-cell lymphoma 2 (Bcl-2) protein regulates programmed cell death throughout the disease conditions by upholding apoptotic pathways. However, the mechanism by which it’s expressed in chondrocytes still needs to be studied in chondrocyte-related disorders. Additionally, explor...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-01-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-023-01377-w |
| _version_ | 1797363053043384320 |
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| author | Muhammad Fakhar-e-Alam Kulyar Quan Mo Wangyuan Yao Yan Li Shah Nawaz Kyein San Loon Ahmed Ezzat Ahmed Aiman A. Alsaegh Khalid M. Al Syaad Muhammad Akhtar Zeeshan Ahmad Bhutta Jiakui Li Desheng Qi |
| author_facet | Muhammad Fakhar-e-Alam Kulyar Quan Mo Wangyuan Yao Yan Li Shah Nawaz Kyein San Loon Ahmed Ezzat Ahmed Aiman A. Alsaegh Khalid M. Al Syaad Muhammad Akhtar Zeeshan Ahmad Bhutta Jiakui Li Desheng Qi |
| author_sort | Muhammad Fakhar-e-Alam Kulyar |
| collection | DOAJ |
| description | Abstract Background The B-cell lymphoma 2 (Bcl-2) protein regulates programmed cell death throughout the disease conditions by upholding apoptotic pathways. However, the mechanism by which it’s expressed in chondrocytes still needs to be studied in chondrocyte-related disorders. Additionally, exploring the potential therapeutic role of Chlorogenic acid (CGA) in confluence with Bcl-2 modulation is of significant interest. Methods In vivo and in vitro studies were performed according to our previous methodologies. The chondrocytes were cultured in specific growth media under standard conditions after expression verification of different microRNAs through high-throughput sequencing and verification of Bcl-2 involvement in tibial growth plates. The effect of Bcl-2 expression was investigated by transfecting chondrocytes with miR-460a, siRNA, and their negative controls alone or in combination with CGA. The RNA was extracted and subjected to a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Western blot analysis and immunofluorescence assays were performed to visualize the intracellular localization of Bcl-2 and associated proteins related to apoptotic and inflammasome pathways. Moreover, apoptosis through flow cytometry was also performed to understand the modulation of concerning pathways. Results The suppression of Bcl-2 induced higher apoptosis and mitochondrial dysfunction, leading to IL-1β maturation and affecting the inflammasome during chondrocyte proliferation. Conversely, overexpression attenuated the activation, as evidenced by reduced caspase activity and IL-1β maturation. In parallel, CGA successfully reduced siRNA-induced apoptosis by decreasing Cytochrome C (Cyto C) release from the mitochondria to the cytoplasm, which in turn decreased Caspase-3 and Caspase-7 cleavage with Bcl-2-associated X protein (Bax). Furthermore, siBcl-2 transfection and CGA therapy increased chondrocyte proliferation and survival. The CGA also showed a promising approach to maintaining chondrocyte viability by inhibiting siRNA-induced apoptosis. Conclusions Targeting Bcl-2-mediated regulation might be a possible treatment for chondrocyte-related conditions. Moreover, these results add knowledge of the complicated processes underlying chondrocyte function and the pathophysiology of related diseases, highlighting the significance of target specific therapies. Video Abstract |
| first_indexed | 2024-03-08T16:15:16Z |
| format | Article |
| id | doaj.art-677dd173f641472184ff39094ab90824 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-03-08T16:15:16Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-677dd173f641472184ff39094ab908242024-01-07T12:36:39ZengBMCCell Communication and Signaling1478-811X2024-01-0122111910.1186/s12964-023-01377-wModulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acidMuhammad Fakhar-e-Alam Kulyar0Quan Mo1Wangyuan Yao2Yan Li3Shah Nawaz4Kyein San Loon5Ahmed Ezzat Ahmed6Aiman A. Alsaegh7Khalid M. Al Syaad8Muhammad Akhtar9Zeeshan Ahmad Bhutta10Jiakui Li11Desheng Qi12College of Veterinary Medicine, Huazhong Agricultural UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityBiology Department, College of Science, King Khalid UniversityDepartment of Laboratory Medicine, Faculty of Applied Medical Sciences, Umm Al-Qura UniversityBiology Department, College of Science, King Khalid UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityLaboratory of Veterinary Immunology and Biochemistry, College of Veterinary Medicine, Chungbuk National UniversityCollege of Veterinary Medicine, Huazhong Agricultural UniversityDepartment of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural UniversityAbstract Background The B-cell lymphoma 2 (Bcl-2) protein regulates programmed cell death throughout the disease conditions by upholding apoptotic pathways. However, the mechanism by which it’s expressed in chondrocytes still needs to be studied in chondrocyte-related disorders. Additionally, exploring the potential therapeutic role of Chlorogenic acid (CGA) in confluence with Bcl-2 modulation is of significant interest. Methods In vivo and in vitro studies were performed according to our previous methodologies. The chondrocytes were cultured in specific growth media under standard conditions after expression verification of different microRNAs through high-throughput sequencing and verification of Bcl-2 involvement in tibial growth plates. The effect of Bcl-2 expression was investigated by transfecting chondrocytes with miR-460a, siRNA, and their negative controls alone or in combination with CGA. The RNA was extracted and subjected to a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Western blot analysis and immunofluorescence assays were performed to visualize the intracellular localization of Bcl-2 and associated proteins related to apoptotic and inflammasome pathways. Moreover, apoptosis through flow cytometry was also performed to understand the modulation of concerning pathways. Results The suppression of Bcl-2 induced higher apoptosis and mitochondrial dysfunction, leading to IL-1β maturation and affecting the inflammasome during chondrocyte proliferation. Conversely, overexpression attenuated the activation, as evidenced by reduced caspase activity and IL-1β maturation. In parallel, CGA successfully reduced siRNA-induced apoptosis by decreasing Cytochrome C (Cyto C) release from the mitochondria to the cytoplasm, which in turn decreased Caspase-3 and Caspase-7 cleavage with Bcl-2-associated X protein (Bax). Furthermore, siBcl-2 transfection and CGA therapy increased chondrocyte proliferation and survival. The CGA also showed a promising approach to maintaining chondrocyte viability by inhibiting siRNA-induced apoptosis. Conclusions Targeting Bcl-2-mediated regulation might be a possible treatment for chondrocyte-related conditions. Moreover, these results add knowledge of the complicated processes underlying chondrocyte function and the pathophysiology of related diseases, highlighting the significance of target specific therapies. Video Abstracthttps://doi.org/10.1186/s12964-023-01377-wApoptosisB-cell lymphoma 2Bcl-2Interleukin-1βInflammasome |
| spellingShingle | Muhammad Fakhar-e-Alam Kulyar Quan Mo Wangyuan Yao Yan Li Shah Nawaz Kyein San Loon Ahmed Ezzat Ahmed Aiman A. Alsaegh Khalid M. Al Syaad Muhammad Akhtar Zeeshan Ahmad Bhutta Jiakui Li Desheng Qi Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid Cell Communication and Signaling Apoptosis B-cell lymphoma 2 Bcl-2 Interleukin-1β Inflammasome |
| title | Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid |
| title_full | Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid |
| title_fullStr | Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid |
| title_full_unstemmed | Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid |
| title_short | Modulation of apoptosis and Inflammasome activation in chondrocytes: co-regulatory role of Chlorogenic acid |
| title_sort | modulation of apoptosis and inflammasome activation in chondrocytes co regulatory role of chlorogenic acid |
| topic | Apoptosis B-cell lymphoma 2 Bcl-2 Interleukin-1β Inflammasome |
| url | https://doi.org/10.1186/s12964-023-01377-w |
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