Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress
This study was carried out to investigate the effects of trehalose (Tre) on antioxidant capacity, endoplasmic reticulum stress (ERS) response and apoptosis of heat-stressed intestinal porcine epithelial cells (IPEC-J2). IPEC-J2 cells were cultured at 37 °C until the end of the experiment (control, C...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-08-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/12/16/2093 |
| _version_ | 1797432550800490496 |
|---|---|
| author | Fan Mo Xu Zhou Mengting Yang Leyi Chen Zhining Tang Chong Wang Yanjun Cui |
| author_facet | Fan Mo Xu Zhou Mengting Yang Leyi Chen Zhining Tang Chong Wang Yanjun Cui |
| author_sort | Fan Mo |
| collection | DOAJ |
| description | This study was carried out to investigate the effects of trehalose (Tre) on antioxidant capacity, endoplasmic reticulum stress (ERS) response and apoptosis of heat-stressed intestinal porcine epithelial cells (IPEC-J2). IPEC-J2 cells were cultured at 37 °C until the end of the experiment (control, CON); exposed to heat stress for 2 h (43 °C, HS); or pretreated with 0.1, 1, 5, 10, and 15 mM trehalose at 37 °C for 4 h prior to heat stress exposure for 2 h. The optimum level of trehalose for protecting against HS-induced cell injuries was determined to be 10 mM, as evidenced by the highest cellular viability and lowest malondialdehyde (MDA) content and lactate dehydrogenase (LDH) activity. Based on these, IPEC-J2 cells were divided into three groups: the first group was cultured at 37 °C until the end of the experiment (control, CON); the second group was exposed to heat stress for 2 h (43 °C, HS); the third group was pretreated with 10 mM trehalose for 4 h at 37 °C prior to heat stress exposure for 2 h (Tre + HS). The reactive oxygen species (ROS) content, superoxide dismutase (SOD) activity, mitochondrial membrane potential (MMP) changes, and expressions of the manganese superoxide dismutase (SOD2), ERS and apoptosis-related proteins were determined. Compared to the CON group, HS significantly increased ROS generation (<i>p</i> < 0.01), decreased SOD activity (<i>p</i> < 0.05), and downregulated protein expression of SOD2 (<i>p</i> < 0.01). Compared to the HS group, Tre supplementation reduced ROS levels and increased SOD activity and SOD2 expression to the levels that were comparable to the control (<i>p</i> < 0.05). The HS-induced ERS response was evidenced by the increased protein expressions of glucose-regulated protein 78 (GRP78) (<i>p</i> < 0.01), eukaryotic translation initiation factor 2α (p-eif2α) (<i>p</i> < 0.01), transcription activator 4 (ATF4) (<i>p</i> < 0.01), and the protein expression of C/EBP homologous protein (CHOP) (<i>p</i> < 0.01), which were the four hallmarks of ERS. The Tre + HS group showed lower expressions of GRP78 (<i>p</i> < 0.01), p-eif2α (<i>p</i> < 0.01), ATF4 (<i>p</i> < 0.01), and CHOP (<i>p</i> < 0.01) than that of the HS group. Tre pretreatment attenuated HS-induced mitochondrial apoptosis in IPEC-J2 cells, demonstrated by the increased MMP and decreased proapoptotic proteins active caspase 3, Bax, and cytochrome c (Cyt c). Taken together, trehalose can protect against HS-induced oxidative damage and endoplasmic reticulum stress-mediated apoptosis in IPEC-J2 cells. These data may provide a nutritional strategy for alleviating heat stress in pig production. |
| first_indexed | 2024-03-09T10:02:10Z |
| format | Article |
| id | doaj.art-1642289727494c0c9463c1ded734099a |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T10:02:10Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-1642289727494c0c9463c1ded734099a2023-12-01T23:19:01ZengMDPI AGAnimals2076-26152022-08-011216209310.3390/ani12162093Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat StressFan Mo0Xu Zhou1Mengting Yang2Leyi Chen3Zhining Tang4Chong Wang5Yanjun Cui6Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaKey Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, China-Australian Joint Laboratory for Animal Health Big Data Analytics, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Zhejiang A&F University, Lin’an, Hangzhou 311300, ChinaThis study was carried out to investigate the effects of trehalose (Tre) on antioxidant capacity, endoplasmic reticulum stress (ERS) response and apoptosis of heat-stressed intestinal porcine epithelial cells (IPEC-J2). IPEC-J2 cells were cultured at 37 °C until the end of the experiment (control, CON); exposed to heat stress for 2 h (43 °C, HS); or pretreated with 0.1, 1, 5, 10, and 15 mM trehalose at 37 °C for 4 h prior to heat stress exposure for 2 h. The optimum level of trehalose for protecting against HS-induced cell injuries was determined to be 10 mM, as evidenced by the highest cellular viability and lowest malondialdehyde (MDA) content and lactate dehydrogenase (LDH) activity. Based on these, IPEC-J2 cells were divided into three groups: the first group was cultured at 37 °C until the end of the experiment (control, CON); the second group was exposed to heat stress for 2 h (43 °C, HS); the third group was pretreated with 10 mM trehalose for 4 h at 37 °C prior to heat stress exposure for 2 h (Tre + HS). The reactive oxygen species (ROS) content, superoxide dismutase (SOD) activity, mitochondrial membrane potential (MMP) changes, and expressions of the manganese superoxide dismutase (SOD2), ERS and apoptosis-related proteins were determined. Compared to the CON group, HS significantly increased ROS generation (<i>p</i> < 0.01), decreased SOD activity (<i>p</i> < 0.05), and downregulated protein expression of SOD2 (<i>p</i> < 0.01). Compared to the HS group, Tre supplementation reduced ROS levels and increased SOD activity and SOD2 expression to the levels that were comparable to the control (<i>p</i> < 0.05). The HS-induced ERS response was evidenced by the increased protein expressions of glucose-regulated protein 78 (GRP78) (<i>p</i> < 0.01), eukaryotic translation initiation factor 2α (p-eif2α) (<i>p</i> < 0.01), transcription activator 4 (ATF4) (<i>p</i> < 0.01), and the protein expression of C/EBP homologous protein (CHOP) (<i>p</i> < 0.01), which were the four hallmarks of ERS. The Tre + HS group showed lower expressions of GRP78 (<i>p</i> < 0.01), p-eif2α (<i>p</i> < 0.01), ATF4 (<i>p</i> < 0.01), and CHOP (<i>p</i> < 0.01) than that of the HS group. Tre pretreatment attenuated HS-induced mitochondrial apoptosis in IPEC-J2 cells, demonstrated by the increased MMP and decreased proapoptotic proteins active caspase 3, Bax, and cytochrome c (Cyt c). Taken together, trehalose can protect against HS-induced oxidative damage and endoplasmic reticulum stress-mediated apoptosis in IPEC-J2 cells. These data may provide a nutritional strategy for alleviating heat stress in pig production.https://www.mdpi.com/2076-2615/12/16/2093heat stressendoplasmic reticulum stressapoptosistrehaloseIPEC-J2 cells |
| spellingShingle | Fan Mo Xu Zhou Mengting Yang Leyi Chen Zhining Tang Chong Wang Yanjun Cui Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress Animals heat stress endoplasmic reticulum stress apoptosis trehalose IPEC-J2 cells |
| title | Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress |
| title_full | Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress |
| title_fullStr | Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress |
| title_full_unstemmed | Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress |
| title_short | Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress |
| title_sort | trehalose attenuates oxidative stress and endoplasmic reticulum stress mediated apoptosis in ipec j2 cells subjected to heat stress |
| topic | heat stress endoplasmic reticulum stress apoptosis trehalose IPEC-J2 cells |
| url | https://www.mdpi.com/2076-2615/12/16/2093 |
| work_keys_str_mv | AT fanmo trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT xuzhou trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT mengtingyang trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT leyichen trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT zhiningtang trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT chongwang trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress AT yanjuncui trehaloseattenuatesoxidativestressandendoplasmicreticulumstressmediatedapoptosisinipecj2cellssubjectedtoheatstress |