ABSTRACT
To observe the protective effect of scutellarin ethyl ester on focal cerebral ischemia injury induced by middle cerebral artery occlusion in rats(MCAOR), and explore its mechanism. Totally 84 male SD rats were randomly divided into seven groups： sham-operated group, model group,positive drug group(niomdipine,12 mg•kg⁻¹), Brevisapin tablets group(48 mg•kg⁻¹), and high, middle and low-dose scutellarin ethyl ester groups(100, 50, 25 mg•kg⁻¹). The MCAOR model was prepared by using thread embolism method to observe the neurological function of rats, the area of cerebral infarction was measured with TTC, and the levels of MDA, SOD and NO in serum were detected with semiautomatic biochemistry analyzer.Ox-LDL and TNF-α cell injury models was established by treating HUVECs with 200 mg•L⁻¹ ox-LDL and 100 μg•L⁻¹ TNF-α,and the levels of MDA, SOD, NO, ET, 6-keto-PGF1α,TXB2, IL-1, IL-6, IL-8, ICAM-1 and PECAM-1 in the cell supernatant were determined. The results showed that scutellarin ethyl ester could effectively improve the neurological function of MCAOR rats, and significantly reduce the area of cerebral infarction. Compared with the model group, activities of SOD and NO in serum increased, while content of MDA decreased. In the cell supernatant, activities of SOD, 6-keto-PGF1α and NO increased, content of IL-1, IL-6, IL-8, ICAM-1, PECAM-1, TXB2, ET and MDA decreased, which indicated that scutellarin ethyl ester has a certain protective effect on focal cerebral ischemia injury induced by middle cerebral artery occlusion in rats, and its mechanism may be related to antioxidative stress, improvement of endothelial function and reduction in inflammatory reaction.
ABSTRACT
OBJECTIVE: To establish an analytical method for simultaneous determination of scutellarin ethyl ester and its related substances by RP-HPLC and identify the main related substances by UPLC-MS. METHODS: The assay was performed on a Kromasil C18 column (4.6 mm×250 mm, 5 μm),and gradient elution was used with methanol-water containting 0.2% formic acid. The flow rate was 1.0 mL·min-1 and the detection wave length was set at 335 nm. The structure of the main related substance was identified by high resolution MS. RESULTS: Under the separation condition, the related substances were completely separated from the principal components. The calibration curve of scutellarin ethyl ester showed good linearity in the concentration range of 5.005-1001 mg·L-1 (r2=0.9999). The average recovery was 99.6%. The main related substance was identified as scutellarin by LC-MS. The contents of total impurities and scutellarin were 1.14% and 0.89%, respectively. CONCLUSION: The method is simple, efficient and accurate, and can be used for the quality control of scutellarin ethyl ester.