1. Academic Validation
  2. PI3K/AKT/mTOR, NF-κB and ERS pathway participated in the attenuation of H2O2-induced IPEC-J2 cell injury by koumine

PI3K/AKT/mTOR, NF-κB and ERS pathway participated in the attenuation of H2O2-induced IPEC-J2 cell injury by koumine

  • J Ethnopharmacol. 2022 Dec 15;116028. doi: 10.1016/j.jep.2022.116028.
Zhihang Yuan 1 Mengran Yang 1 Zengenni Liang 2 Chenglin Yang 1 Xiangyi Kong 1 You Wu 1 Siqi Wang 1 Hui Fan 1 Can Ning 1 Wenguang Xiao 1 Zhiliang Sun 3 Jing Wu 4
Affiliations

Affiliations

  • 1 Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, PR China; Hunan Co-innovation Center of Animal Production Safety, Changsha, 410128, PR China.
  • 2 Department of Hunan Agricultural Product Processing Institute, Hunan Academy of Agricultural Sciences, Changsha, 410128, PR China.
  • 3 Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, PR China; Hunan Co-innovation Center of Animal Production Safety, Changsha, 410128, PR China; Hunan Engineering Research Center of Veterinary Drug, Hunan Agricultural University, Changsha, 410128, PR China; National Research Center of Engineering and Technology for Utilization of Botanical Functional Ingredients, PR China. Electronic address: zhiliangsun15@163.com.
  • 4 Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, PR China; Hunan Co-innovation Center of Animal Production Safety, Changsha, 410128, PR China. Electronic address: wujing@hunau.edu.cn.
Abstract

Ethnopharmacological relevance: Koumine, an indole alkaloid extracted from Gelsemium elegans Benth, exerts anti-inflammation and antioxidant activities. However, the effects of koumine on intestinal injury induced by H2O2 and its potential molecular mechanisms need larger studies.

Aim of the study: We established an IPEC-J2 cell damage model induced by H2O2 to explore the protective mechanism of koumine on intestinal injury.

Materials and methods: In the experiment, cell damage models were made with hydrogen peroxide. To assess the protective effect of koumine on H2O2-induced IPEC-J2 cell injury, CCK-8, the release of LDH and ROS, transmission electron microscopy and Annexin V-FITC/PI were employed. Western Blot and Quantitative Real-Time PCR were used to determine the potential alleviated mechanism of koumine on H2O2-trigged IPEC-J2 cell damage.

Results: The results of CCK-8 and LDH implied that koumine has a mitigative effect on H2O2-induced cell damage via upregulating cell viability and suppressing cell membrane fragmentation. Simultaneously, koumine notably inhibited the level of pro-inflammatory factors (IL-1β, IL-6, IL-8, TNF-α and TGF-β), the over-production of ROS along with decreasing the injury of mitochondrion, endoplasmic reticulum and lysosome induced by H2O2. Moreover, koumine dramatically attenuated H2O2-triggered IPEC-J2 cell Apoptosis and Autophagy. Subsequently, Western blot analysis identified NF-κB, PI3K and ERS as possible pathway responsible for the protective effect of koumine on H2O2-stimulated IPEC-J2 cell inflammation.

Conclusions: This in vitro experimental study suggests that koumine suppresses the H2O2-induced activation of inflammatory pathways, oxidative injury, ER stress, Apoptosis and Autophagy, which provide a rationale for therapeutically use in major intestinal diseases.

Keywords

Apoptosis; Autography; ERS; IPEC-J2; Inflammation; Koumine; NF-κB; PI3K; ROS.

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