Evaluation of Anti-inflammatory Activity of Ethanol Extracts ofRadix Sophorae Flavescentis, Radix Arnebiae and Radix Scrophulariae by Zebrafish Model
LIU Ming1, GONG Qingfang1, LU Xin1,2, HU Shuo1, LIANG Mingzhong1, WU Yingrui1, CAI Xiaohui1
1. Guangxi Key Laboratory of Marine Environmental Disaster Process and Ecological Protection Technology, CollegeofMarineSciences, Beibu Gulf University, Qinzhou 535011, China; 2. College of Marine Fisheries, Fangchenggang Vocational and Technical College, Fangchenggang 538000, China
Abstract：In order to evaluate the anti-inflammatory activity of ethanol extracts of Radix Sophorae Flavescentis, Radix Arnebiae and Radix Scrophulariae [extracted by ethanol reflux method at 80 ℃, with solvent recovered; dissolved in dimethyl sulfoxide (DMSO) before use] using a transgenic zebrafish Danio rerio inflammation model, transgenic Tg zebrafish embryos were raised at water temperature of(26±1)℃ and moved into 0.2 mmol/L phenylthiourea (PTU) embryo medium when developed to 22—26 hours post-fertilization. After 12 h, the medium was discarded, embryos were washed 1—2 times, distilled water was added, and the embryos were returned to the 28 ℃ temperature-controlled culture and breeding system to raise the larvae to 3 days post-fertilization. Zebrafish caudal fin injury inflammation and copper sulfate inflammation models (soaked in 13.3 μmol/L copper sulfate solution for 2 h) were established. The larvae were then placed in distilled water (blank control group), 1 mg/mL Radix Sophorae Flavescentis decoction (Radix Sophorae Flavescentis group), 1 mg/mL Radix Arnebiae decoction (Radix Arnebiae group), 1 mg/mL Radix Scrophulariae decoction (Radix Scrophulariae group), and 90 μg/mL dexamethasone solution for 2 h (DEX group). Next, blue light (415—455 nm) was used to excite the green fluorescence of neutrophils in vivo in the transgenic Tg zebrafish (mop-GFP), and the distribution and number of neutrophils in the tail were observed under a fluorescence microscope, and the expression levels of TNF-α, IL-6, and IL-10 genes to evaluate the anti-inflammatory activity of the various traditional Chinese medicines by Real-time quantitative PCR. The results showed that there was the fluorescent neutrophils migration average of 45.7±1.5 in the tail inflammation model of zebrafish, 12.2±1.4 in the Radix Sophorae Flavescentis group, 9.3±0.7 in the Radix Arnebiae group, 13.3±1.0 in the Radix Scrophulariae group and 23.2±0.4 in the DEX group, with average migration of 25.5±1.2 in the copper sulfate inflammation model of zebrafish, 2.5±1.1 in the Radix Sophorae Flavescentis group, 4.8±0.9 in the Radix Arnebiae group, 4.5±0.3 in the Radix Scrophulariae group and 4.3±0.7 in the DEX group. The migration averages of fluorescent neutrophils were shown to be significantly decreased in the DEX group, the Radix Sophorae Flavescentis group, the Radix Arnebiae group, and the Radix Scrophulariae group compared with the inflammation model group. The expression levels of TNF-α and IL-6 genes were significantly decreased compared with the inflammation model group, while the expression level of IL-10 gene was significantly increased after treatment with the herbal extracts (P<0.05). The findings indicate that ethanol extracts of Radix Sophorae Flavescentis, Radix Arnebiae and Radix Scrophulariae have strong anti-inflammatory activities, inhibiting the migration and aggregation of neutrophils in zebrafish, and that can provide reference for the prevention and treatment of fish enteritis with Chinese herbal medicine.
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2026, Vol. 45 
