Background:Anoectochilus roxburghii (Orchidaceae) is a traditional Chinese medicinal herb with anti-inflammatory, antilipemic, liver protective, immunomodulatory, and other pharmacological activities. Kinsenoside (KD), which shows protective effects against a variety types of liver damage, is an active ingredient extracted from A. roxburghii. However, the liver protective effects and potential mechanisms of KD in alcoholic liver disease (ALD) remain unclear. This study aimed to investigate the liver protective activity and potential mechanisms of KD in ALD.Methods: AML12 normal mouse hepatocyte cells were used to detect the protective effect of KD against ethanol-induced cell damage. An alcoholic liver injury model was induced by feeding male C57BL/6J mice with an ethanol-containing liquid diet, in combination with intraperitoneal administration of 5% carbon tetrachloride (CCl₄) in olive oil. Mice were divided into control, model, silymarin (positive control), and two KD groups, treated with different doses. After treatment, hematoxylin–eosin and Masson’s trichrome staining of liver tissues was performed, and serum alanine aminotransferase (ALT) and aspartate transaminase (AST) levels were determined to assess the protective effect of KD against alcoholic liver injury. Moreover, proteomics techniques were used to explore the potential mechanism of KD action, and ELISA assay, immunohistochemistry, TUNEL assay, and western blotting were used to verify the mechanism.Results: The results showed that KD concentration-dependently reduced ethanol-induced lipid accumulation in AML12 cells. In ALD mice model, the histological examination of liver tissues, combined with the determination of ALT and AST serum levels, demonstrated a protective effect of KD in the alcoholic liver injury mice. In addition, KD treatment markedly enhanced the antioxidant capacity and reduced the endoplasmic reticulum (ER) stress, inflammation, and apoptosis compared with those in the model group. Furthermore, KD increased the phosphorylation level of AMP-activated protein kinase (AMPK), inhibited the mechanistic target of rapamycin, promoted the phosphorylation of ULK1 (Ser555), increased the level of the autophagy marker LC3A/B, and restored ethanol-suppressed autophagic flux, thus activating AMPK-dependent autophagy.Conclusion: This study indicates that KD alleviates alcoholic liver injury by reducing oxidative stress and ER stress, while activating AMPK-dependent autophagy. All results suggested that KD may be a potential therapeutic agent for ALD.

Excessive stimulation of hepatotoxins and drugs often lead to acute liver injury, while treatment strategies for acute liver injury have been limited. Methyl 6-O-cinnamoyl-α-d-glucopyranoside (MCGP) is a structure modified compound from cinnamic acid, a key chemical found in plants with significant antioxidant, anti-inflammatory, and antidiabetic effects. In this study, we investigated the effects and underlying mechanisms of MCGP on acetaminophen (APAP)- or carbon tetrachloride (CCl₄)-induced acute liver injury. As a result, MCGP inhibited cell death and apoptosis induced by APAP or CCl₄, and suppressed the reactive oxygen species (ROS) generation stimulated by H₂O₂ in liver AML12 cells. In vivo, MCGP alleviated APAP/CCl₄-induced hepatic necrosis and resumed abnormal aminotransferase activities and liver antioxidase activities. In addition, MCGP depressed APAP- or CCl₄-induced oxidative stress through the suppression of CYP2E1 and activation of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. MCGP also enhanced the number of PCNA-positive hepatocytes, increased hepatic PCNA and Bcl-XL, and decreased BAX expression in APAP-/CCl₄-intoxicated mice. Furthermore, MCGP activated the GSDMD-N/cleaved caspase 1 pathway. In summary, MCGP might act as a potential therapeutic drug against drug-induced and chemical-induced acute liver injuries, and its underlying mechanisms might engage on the pressing of oxidative stress, refraining of hepatocyte apoptosis, and facilitating of liver regeneration.

Background: Studies using biospecimens can help reveal pathogenic mechanisms and improve prevention, diagnosis, and treatment of diseases. However, there is still a lack of relevant investigation data, which can provide initial evidence for establishing or improving relevant laws and regulations, on people's willingness to donate biospecimens, and whether they agree to waive the right of obtaining informed consent in the special period of sudden outbreak of new infectious diseases. Objectives: To investigate people's willingness to donate their remnant biospecimens of clinical tests for research in the context of the COVID-19 pandemic and their willingness to sign the informed consent for research using their biospecimens. Methods: We conducted a survey using an online questionnaire, which included questions on basic personal information, COVID-19-related information, donation of remnant biospecimens, willingness to sign informed consent, and reasons to do so. Results: Among the 721 valid responses, 620 respondents (86.0%) reported that they would be willing to donate their remnant biospecimens for research, of whom 434 (70.0%) reported that they would donate their remnant biospecimens without signing the informed consent. Of the 11 specified influencing factors, occupation, household income, and degree of concern about the COVID-19 pandemic were associated with willingness to donate remnant biospecimens. Gender and age were associated with willingness to donate remnant biospecimens without signing the informed consent. The main reasons for unwillingness to donate remnant biospecimens and sign the informed consent were a limited knowledge of research and privacy concerns. Conclusions: Most respondents reported that they would be willing to donate their remnant biospecimens for biomedical research without signing an informed consent in the context of the COVID-19 pandemic. Lack of understanding of the proposed research and concerns about personal privacy were the main reasons for unwillingness to donate biospecimens and signing the informed consent.