Huang Qi

The QSKL used in this study is manufactured by Beijing university of Chinese medicine (Beijing, China) using the six Chinese herbs at a composition of 460 g Radix Astragali Mongolici, 230 g salvia miltiorrhiza bunge, 160 g Flos Lonicerae, 160 g scrophularia, 140 g Radix Aconiti Lateralis Preparata, and 90 g Radix Glycyrrhizae. Briefly, the residue of Radix Astragali Mongolici is mixed with all salvia miltiorrhiza bunge, Flos Lonicerae, scrophularia, and Radix Glycyrrhizae, follow by extraction with hot water (twice, 2 hr each). The water extract is then concentrated to form a paste, and the ethanol is added for 24 hr, the filtration is collected to form the final product. Based on the recommended daily human dosage of 20 g/d, according to the equivalent conversion between animal and people by body surface area, dosage of 508 mg/kg is chosen in present study.

The broth for Samgyetang was prepared using 180 g of Astragalus membranaceus root, 300 g of Mulberry branch, 276 g of Kalopanax septemlobus branch, 60 g of licorice, 324 g of Siberian ginseng, and 210 g of salt per 36 L of water, followed by boiling for approximately 1 hour. The solids in the broth were removed, and the broth was filtered using a stainless mesh filter (8 mm). The final salt concentration was adjusted to 0.6% using a salinity meter (PAL-03S, Atago Co., Ltd., Tokyo, Japan) and by adding drinking water.
To prepare the stuffing, glutinous rice was soaked for 1 h and rinsed prior to use. Garlic, ginseng, and dried jujube were rinsed with cold water. Approximately 35 g of glutinous rice, 8 g of garlic, 5 g of jujube and 7 g of ginseng were placed in rice paper, which was soaked with warm water, and wrapped. The wrapped rice, breast and thigh meat were then stuffed into a retort pouch (length 19 cm [length]×25 cm [height], polyethylene terephthalate = 16 μm, aluminum = 9 μm, nylon = 15 μm, polypropylene =100 μm) gifted from Sunbong Food (Incheon, Korea) and were filled with broth until the weight reached 1,000±0.05 g. The pouches were then sealed (WB-1150VP; Woobin Tech Co., Ltd., Incheon, Korea). Two extra pouches were used to measure the F0 value during the retorting process. The retort process was performed using a steam-type retort sterilization chamber (Steri-ace, Kyungshan Co., Ltd., Gyeongsan, Korea). The heating process was started at 65°C, and the temperature was raised to 121°C for the first sterilization (pressure = 1.5 kgf/cm²; holding time = 3 min). Next, the temperature was raised again to 122°C for 10 s for the second sterilization (pressure = 1.7 kgf/cm²; holding time = 95 min). The cooling process was held at 1.7 kgf/cm² for 20 min. Retort-pouched Samgyetang was stored in a refrigerator (5°C±0.5°C) prior to analysis (within 3 days). For analysis, 10 pouches of retorted Samgyetang from each treatment group were used for meat quality analysis, while the other 28 pouches from each treatment group were used for sensory evaluation.

Astragalus roots were purchased from Tianjin Tongrentang Group Co., Ltd. (Tianjin, China). Bax, β-actin, Bcl-2, and Cyt c antibodies were purchased from ImmunoWay Biotechnology Company (Plano, TX, USA). Apoptosis index-related detection kits for cell experiments were all provided by Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Human hepatoma HepG2 cells were provided by the Shanghai Institute of Biological Sciences at the Chinese Academy of Sciences (Shanghai, China). All other chemicals used in this study were of analytical grade.

Compound chemical composition and drug targets were collected and predicted. Dangshen (Codonopsis pilosula (Franch.) Nannf), Huangqin (Scutellaria baicalensis Georgi), Baizhu (Atractylodes macrocephala Koidz), Fuling (Wolfiporia cocos (F.A. Wolf) Ryvarden & Gilb), Huangqi (Astragalus membranaceus (Fisch.) Bunge), Yiyiren (Semen Coicis), Wumei (Fructus Mume), Fangfeng (Saposhnikovia divaricata (Turcz.) Schischk), Chenpi (Citrus reticulata Blanco), Niuxi (Achyranthes bidentata Blume), and Gancao (Glycyrrhiza uralensis Fisch) were inputted to the TCMSP database (https://old.tcmsp-e.com/tcmsp.php), to collect all compounds [12 (link)]. Then, according to the pharmacokinetic principle, the suitable compounds are screened based on oral bioavailability (OB) ≥ 30% and drug-like index (DL) ≥ 0.18. Drug target data were obtained from DrugBank (https://go.drugbank.com/) and standardized using the UniProt (https://www.Uniprot.org/) database [13 (link)]. Based on GeneCards (https://www.genecards.org/) and OMIM-NCBI databases (https://www.ncbi.nlm.nih.gov/omim), IBD as keyword was used to screen disease targets, while for species selection “Homosapiens” was used. Draw Venn Diagrams tool in R software is used to process drug targets and disease targets obtained in the above steps to obtain the intersection gene targets and output Venn Diagram for display.
PPI network interaction analysis was performed on the active chemical components and core targets of Qinghua Jianpi Recipe by using the STRING database and Cytoscape3.7.1 software. According to the requirements of Cytoscape, the “source-target” data table was constructed, and the NetworkAnalyzer plug-in was used to construct the regulation network of TCM, active component, and target. In the generated regulation network, nodes represent the interaction between TCM, active component, and target, and edges represent the interaction between active the component and target disease. Furthermore, MCC algorithm of CytoHubba plug-in was used to calculate and construct PPI network of each target. Based on Bioconductor (https://www.bioconductor.org/) in the R package (https://www.r-project.org/) and clusterProfiler 3.12.0 to GO (gene ontology) core target function and KEGG pathway enrichment analysis (KEGG pathway analysis), Homo sapiens was selected and a threshold P < 0.05 was set. According to the results, the core target-critical pathway network was conducted in Cytoscape 3.7.1 software.