ABSTRACT
Aim To investigate the effects of Kudino- side D on lipid accumulation induced by oxidized low density lipoprotein ( ox-LDL) and inflammation induced by lipopolysaccharide ( LPS ) in RAW264.7 cells.Methods Foam cells were established by incubating the RAW264.7 cells with ox-LDL.The concentration of lipid droplets in the cells was observed by oil red staining, and the level of total cholesterol (TC) in cells was measured by enzyme method.The gene and protein expressions of scavenger receptors CD36 and SR-A1, ATP binding cassette transporters A1 and Gl ( ABCA1 and ABCGI) were detected by RT-qPCR and Western blot, respectively.The expressions of inter- leukin-6 (IL-6), interleukin-1 (3 (IL-ip), monocyte chemoattractant protein-1 (MCP-1 ) and tumor necrosis factor-a (TNF-a) were detected by ELISA and RT-qPCR.The protein expressions of mTOR and p-mTOR were detected by Western blot.Results Compared with model group, the high dose of Kudinoside D decreased the content of TC and down-regulated the gene and protein expression of SR-A induced by ox-LDL.Meanwhile Kudinoside D also decreased the levels of IL-ip and MCP-1 and down-regulated the protein expression of p-mTOR induced by LPS.Conclusions Kudinoside D may reduce the intracellular TC content by down-regulating the gene and protein expression of SR-A1.Kudinoside D may play an anti-inflammatory role through mTOR pathway.
ABSTRACT
Objective To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of six components in Ilex kudingcha by HPLC. Methods The purity of kaempferol-3-O-β-D-rutanoside, isorhamnetin-3-O-β-D-rutanoside, kudinoside C, kudinoside A, kudinoside E, and kudinoside D was determined by HPLC-MS in test sample taking kudinoside C as internal standard, relative correction factors of the other five compounds were determined, and their contents were calculated. At the same time, the contents of these six components were determined by external standard method (ESM), and the difference between ESM and QAMS was compared to verify the feasibility and accuracy of QAMS method. Results There was no significant difference of the six components’ contents calculated between QAMS method and ESM method. Conclusion It is convenient, feasible, and accurate to measure the content of six components in I. kudingcha by QASM method.
ABSTRACT
Objective: To establish an HPLC method for simultaneously determining latifoloside G, latifoloside H, kudinoside G, latifoloside C, kudinoside A, kudinoside E, and kudinoside D in Ilex kudingcha. Methods: The separation was performed on a YMC C18 column (YMC-pack ODS-A, 250 mm × 4.6 mm, 5 μm) with water-acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was 210 nm and the column temperature was 30℃. Results: Seven triterpenoid saponins all have good separating degree, good linear range, and linearity (r > 0.999 5). The average sample recovery rates were 99.25%-100.8%, RSD < 2%. Conclusion: The established method is rapid, accurate, and with has repeatability and stability, which could provides the scientific evidence for the quality control of I. kudingcha.