R signaling by redox homeostasis by way of the detoxification of ROS [17]. LKB1 also can regulate NF-kB-mediated macrophage activation. LKB1-deficient macrophages show greater production and expression of proinflammatory cytokines just after lipopolysaccharide (LPS) stimulation and enhanced NF-kB activity [18]. Some reports indicate that CaMKK participates in NF-B-mediated inflammatory signals, which may be involved in the proinflammatory activity of macrophages. As a result, CaMKK/LKB1/AMPK may very well be involved in liver pathology, because inhibiting its activity can protect against the occurrence of liver illnesses. The mechanism by which paracetamol causes liver damage continues to be unclear, but escalating evidence shows that mediators of oxidative pressure and inflammation are involved. Sanghuangporus sanghuang (SS), a rare and valuable medicinal fungus, has been made use of to treat numerous ailments in Taiwan, China, Japan, and Korea for hundreds of years [19,20]. In traditional usage, SS has been applied for the therapy of diarrhea, night sweats, hemorrhoids, eczema, stomach discomfort, bleeding, vaginal discharge, and amenorrhea. The pharmacological effects of SS are reported to include antioxidant, anti-inflammatory, anti-tumor, anti-aging, anti-diabetic, and antiviral effects [213]. The scientific name “S. sanghuang” was controversial just before 2018; data then showed that Phellinus linteus or Inonotus Sanghuang was not the right name and that it must be S. sanghuang, for the reason that a new species that only grew on reside mulberry trees had been discovered. Information suggested that SS could effectively block oxidative anxiety and inflammatory responses in paracetamol-induced liver injury and that its mechanism may possibly be related towards the MAPK/NF-B, Keap1/Nrf2/HO-1, TLR4/PI3K/Akt, and CaMKK/LKB1/AMPK pathways. two. Components and Approaches 2.1. Reagents Paracetamol, NAC, other chemical substances and solvents had been bought from Phospholipase Inhibitor Biological Activity Sigma-Aldrich (St. Louis, MO, USA). ELISA kits for detecting the release of mouse TNF-, IL-1, and IL-6 had been bought from BioLegend Inc. (San Diego, CA, USA). Primary antibodies for Western blotting against the proteins COX-2, p-JNK, catalase, GPx, SOD, CYP2E1, AMPK, LKB1, CaMKK, p-AMPK, p-LKB1, and p-CaMKK were bought from GeneTex (San Antonio, TX, USA). Antibodies against JNK, p-ERK, ERK, p-p38, and p-IB- were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against iNOS, NF-B, IB, p38, HO-1, Nrf-2, and -actin had been purchased from Abcam (Cambridge, UK, USA). Protein assay kits (Bio-Rad Laboratories Ltd., Watford, Herts, UK) were obtained as indicated. two.2. Supply of Material The mycelium of S. sanghuang utilised in this study was fermented by Taiwan Grape King Biological Co., Ltd. (Chung-Li City, Taiwan), and its strain was proposed by Dr. Sheng-Hua Wu from the Angiotensin Receptor Antagonist Source National Museum of Organic Science. The S. sanghuang mycelium originated on the trunk of Morus in Yanping Township, Taitung County, Taiwan. It was collected in 2011/5. The “voucher specimens” are stored within the National Museum of All-natural Science (Wu 1105-1). two.3. Sample Preparation Dried mycelium powder was soaked in 70 ethanol for 1 week, then, the residue was filtered out. The filtrate was concentrated beneath decreased stress to take away the ethanol. This step was repeated three instances to acquire the extract, which was then stored at -20 C. two.4. Animals Six- to seven-week old pathogen-free male ICR mice (physique weights, 205 g) were obtained from BioLASCO Taiwan Co., Ltd. (Taipei, Taiwan). Six mice wer.