Thonningianin A ameliorates acetaminophen-induced liver injury by activating GPX4 and modulating endoplasmic reticulum stress

Lai Shanglei ¹ Yingyan Ye ² Qinchao Ding ³ Xiaokai Hu ⁴ Fu Ai ⁵ Lan Wu ⁴ Wenjing Cao ³ Qingsheng Liu ⁶ Xiaobing Dou ⁴ Xuchen Qi ^7*

• ¹ Department of Medical Research Center, Shaoxing People’s Hospital, Shaoxing, Zhejiang, China
• ² The First People’s Hospital of Hangzhou Linan District, Hangzhou, Jiangsu Province, China
• ³ School of Public Health, Zhejiang Chinese Medical University, Hangzhou, Jiangsu Province, China
• ⁴ College of Life Science, Zhejiang Chinese Medical University, Hangzhou, Zhejiang Province, China
• ⁵ Hangzhou Xixi Hospital, Hangzhou, Zhejiang Province, China
• ⁶ Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang Province, China
• ⁷ Department of Neurosurgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China

Acetaminophen (APAP) is widely used as an analgesic and antipyretic; however, overdose can lead to acute liver injury (ALI), representing a significant challenge for public health due to limited treatment options. Current research highlights the need for safer and more effective therapies for APAP-induced liver injury, especially those that target oxidative and endoplasmic reticulum (ER) stress pathways. This study investigates the protective effects of Thonningianin A (TA), a flavonoid compound derived from Penthorum chinense Pursh, in mitigating APAP-induced hepatotoxicity. The experimental design involved administering TA at doses of 20 mg/kg and 40 mg/kg to C57BL/6 mice prior to inducing hepatotoxicity with APAP. TA treatment significantly lowered plasma ALT and AST levels, inhibited the production of inflammatory cytokines, and reduced oxidative stress markers in liver tissues. Furthermore, TA modulated apoptosis-related proteins by increasing BCL-2 expression while decreasing CHOP and BAX levels. It alleviated endoplasmic reticulum (ER) stress by downregulating GRP78, p-PERK, and ATF4. Notably, liver-specific GPX4 knockdown, achieved through AAV-8-mediated shRNA delivery, abolished the hepatoprotective effects of TA, underscoring GPX4's essential role in mediating TA-induced hepatoprotection. These findings suggest TA as a promising therapeutic agent in managing APAP-induced liver injury, with its unique action on both oxidative and ER stress pathways contributing to its hepatoprotective efficacy.