Hyp kit

The liver tissues were embedded and sectioned for H&E and Masson staining [12 (link)] using staining kits purchased from Beijing Solarbio Science & Technology Co., Ltd. (catalog no. G1120, G1346, respectively).
Films of H&E and Masson staining were read under an Olympus BX51 microscope to assess the egg granulomatous lesions and hepatic fibrosis in each group. Twenty-five single egg granulomas with clear and well-separated edges were randomly selected from each group. The average areas of the single granuloma and fibrosis were calculated using Cellsens Dimension software.
The content of hydroxyproline (HYP) in the liver tissues of mice was determined following the HYP kit (no. A030-2-1, Nanjing Jiancheng Bioengineering Institute) manufacturer's instructions.

Astragaloside IV (purity ≥98%) and ferulic acid (purity≥98%) were bought from Shanghai yuan ye Bio-Technology Co., Ltd. (Shanghai, China). MIR-29 b agomir and agomir negative control were bought from Guangzhou Ribobio Co., Ltd. (Guangzhou, China). The assay kits of MDA, ROS, and SOD for mice were purchased from Jiancheng Bioengineering Institute (Nanjing, China). The HYP kit was bought from Nanjing Jiancheng Bioengineering Co., Ltd. (Nanjing, China).

Mice were anesthetized with 2% isoflurane and subjected to cardiac perfusion to remove blood from the lungs. Approximately 100 mg of lung tissue sample (stored at −80°C) was hydrolyzed by adding 1 mL of hydrolysate and incubated at 95°C for 20 min (the samples were mixed again after 10 min of incubation). According to the manufacturer’s instructions, an HYP kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) was used to measure the amount of HYP in the lung tissue.

To quantify lung collagen contents, the hydroxyproline (HYP) contents of the lungs were measured with a HYP kit (Nanjing Jian Cheng Institute, China) according to the manufacturer's instruction. The HYP contents were expressed as micrograms per gram (μg/g) of lung wet weight.

Arctigenin was obtained from Nanjing Zelang Medical Technology Co. Ltd. (Nanjing, China). Paraquat dichloride was purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against the following proteins were used: Occludin, α-SMA, Wnt3a, β-Catenin, GSK-3β, P-GSK-3β, β-actin (Abcam, United States). Cytokeratin 18(Proteintech group Inc., United States), E-cadherin, Vimentin (Cell Signaling Technology, Beverly, MA, United States), Horseradish peroxidase (HRP)-conjugated goat anti-mouse and goat anti-rabbit IgG were obtained from Santa Cruz Biotechnogy (Santa Cruz, CA, United States). HYP kit (Nanjing Jiancheng Bioengineering Institute) Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were provided by Gibco (Rockville, MD, United States). All other chemicals and reagents in this study were of analytical grade.

The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hydroxyproline (HYP) in serum were assessed using ALT Kit (#C009‐2–1, Jiancheng, China), AST Kit (#C010‐2–1, Jiancheng, China) and HYP Kit (#A030‐2–1, Jiancheng, China) according to manufacturer's instructions.