Total triterpenes of Euphorbium alleviates rheumatoid arthritis via Nrf2/HO-1/GPX4 pathway / 中国中药杂志

This study aims to investigate the effect and mechanism of total triterpenes of Euphorbium in treating rheumatoid arthritis(RA). The rat model of RA was established with Freund's complete adjuvant(FCA). Male rats were randomly assigned into control, model, Tripterygium glycosides(7.5 mg·kg~(-1)), and low-, medium-, and high-dose total triterpenes of Euphorbium(32, 64, and 128 mg·kg~(-1), respectively) groups, with 10 rats in each group. In other groups except the control group, 0.2 mL FCA was injected into the right hind toe. Rats in the intervention groups were administrated with corresponding drugs by gavage, and the control group and the model group with the same volume of 0.5% CMC-Na solution once a day. During the treatment period, the swelling degree of the hind paw was measured and the arthritis was scored until day 30. At the end of drug administration, the pathological changes of the joint tissue were observed by hematoxylin-eosin staining. The content of malondialdehyde(MDA), glutathione(GSH), and Fe~(2+) and the activity of superoxide dismutase(SOD) in the joint tissue were measured by biochemical colorimetry. RT-PCR was performed to determine the mRNA levels of nuclear factor erythroid 2-related factor 2(Nrf2), glutathione peroxidase 4(GPX4), and acyl-CoA synthetase long chain family member 4(ACSL4) in the joint tissue. Western blot was employed to determine the protein levels of Nrf2, Kelch-like ECH-associated protein 1(Keap1), heme oxygenase-1(HO-1), NAD(P)H quinone oxidoreductase 1(NQO1), SOD2, GPX4, and ACSL4 in the joint tissue. The results showed that the treatment with Tripterygium glycosides(7.5 mg·kg~(-1)) and total triterpenes of Euphorbium(32, 64, and 128 mg·kg~(-1)) alleviated the swelling degree of bilateral hind limbs, decreased the arthritis score, reduced joint tissue lesions and the content of MDA and Fe~(2+) in the joint tissue, and increased GSH content and SOD activity. Furthermore, the interventions up-regulated the protein and mRNA levels of Nrf2 and GPX4, down-regulated the protein and mRNA levels of ACSL4, and up-regulated the protein levels of Keap1, NQO1, HO-1, and SOD2 in the Nrf2/HO-1/GPX4. In summary, the total triterpenes of Euphorbium can treat RA by inhibiting lipid peroxidation and abnormal ferroptosis, which may involve the Nrf2/HO-1/GPX4 signaling pathway.

Content Determination of Tetracyclic Triterpenoids as Euphol and Euphorbol in Euphorbium resinifera and the Optimization of Extraction Technology / 中国药房

OBJECTIVE：To establish a method for the simultaneous determination of euphol and euphorbol in Euphorbium resinifera，and to optimize the extraction technology. METHODS ：HPLC method was adopted. The determination was performed on Agilent ZORBAX Eclipse Plus C 8 column with mobile phase consisted of acetonitrile-water （90∶10，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and column temperature was 30 ℃. The sample size was 10 μL. Using the contents of euphol and euphorbol ，yield of the extract as evaluation index ，comprehensive score was conducted. The extraction technology was optimized by L （9 34）orthogonal tests ，with ethanol volume fraction ，extraction time and solvent dosage as factors. RESULTS：The linear ranges of euphol and euphorbol were 0.030 4-1.216 mg/mL（r＝0.999 6）and 0.01-0.4 mg/mL（r＝0.999 9）， respectively. RSDs of precision ，stability（24 h）and producibility tests were all lower than 2％. Average recoveries were 100.46％ （RSD＝1.03％，n＝6）and 99.36％（RSD＝0.91％，n＝6）. The optimized technology was extracting with 40 mL 95％ ethanol for 1 h. After 3 times of validation tests showed that average content of euphol was 94.43 mg/g（RSD＝0.92％，n＝3），and that of euphorbol was 25.42 mg/g（RSD＝0.98％，n＝3）；average yield of the extract was 51.42％（RSD＝1.95％，n＝3），and average comprehensive score was 98.87（RSD＝0.92％，n＝3）. CONCLUSIONS ：Established method is simple ，accurate and reproducible ， which can be used for the quality control of E. resinifera . The optimized extraction technology is simple and stable.