RESUMO
OBJECTIVE:To investigate the anti-inflammatory activity of 70% ethanol extracts from Garcinia oblongifolia (GOEE)on LPS-induced RAW 264.7 cells and its potential molecular mechanism. METHODS :GOEE was obtained after the fresh G. oblongifolia epicarp refluxed with 70% ethanol. The contents of total phenol and total flavonoids were determined by Folin-Ciocalteau assay and UV spectrophotometer. MTT assay was used to detect the cytotoxicity of different doses of GOEE. The inflammatory model was induced in RAW 264.7 cells by lipopolysa- ccharide (LPS). Using dexamethasone and N-acetyl-L-cysteine as positive control ,Griess assay and 2′,7′-dichloro- fluorescein assay were used to detect the contents of NO in cell culture medium and ROS in cells. The levels of TNF-α,IL-6,and IL- 1β in cell culture medium were measured by ELISA. The protein expression of p 65,p-p65,IκBα,p-IκBα,HO-1 in cells and NRF 2 in nucleus were determined by using Western blotting assay. RESULTS:The contents of total phenol and flavonoids in GOEE were (20.191±1.264)and(12.571±0.020)mg/g,respectively. At the concentration below 500 μ g/mL, GOEE had no significantly effect on survival rate of RAW 264.7 cells(P> 294043)0.05). Compared with control group ,the contents of NO and ROS,the levels of TNF-α,IL-6 and IL- 1β,ratio of p-p 65 top65,ratio of p-IκBα to IκBα,protein expression of NRF 2 were increased significantly in LPS model group (P<0.05 or P<0.01). Compared with LPS model group ,the contents of NO(except for GOEE 50 μg/mL group)and ROS ,the levels of TNF-α,IL-6 and IL- 1β,ratio of p-p 65 to p 65 and ratio of p-IκBα to IκBα were decreased significantly in GOEE groups and positive control groups ,while protein expression of HO- 1 and NRF 2 were increased significantly (P<0.05 or P<0.01). CONCLUSIONS:GOEE attenuates LPS-induced macrophages inflammation injury by inhibiting the inflammatory response and the phosphorylation of NF-κB pathway,promoting NRF 2 protein transportation to the nucleus.
RESUMO
OBJECTIVE:To investigate the correlation of storage life and effective composition content with color value of Carthamus tinctorius ,and to provide reference for the quality evaluation of C. tinctorius with different years of storage. METHODS:Using 24 batches of C. tinctorius from same place of production with different years of storage (0,1,2 years,8 batches each type )as samples ,the contents of hydroxysafflor yellow A (HSYA)and kaempferol were determined by HPLC. Color value [lightness value (L*),red-green value (a*),yellow-blue value (b*)] were determined by spectrophotometer. SPSS 19.0 statistical software was used to analyze the correlation of storage life and effective composition content with color value. RESULTS:Kaempferol content was still high after 1 year or 2 years of storage (0.161%,0.061%,respectively). However ,the content of HSYA decreased with the prolongation of the storage life (the average content of HSYA were 2.46%,1.58%,and 1.51% after storage 0,1 and 2 years,respectively),and the color of the drug became darker (a* value decreased ). Results of correlation analysis showed that the content of HSYA was positively associated with color value L*,a*(r=0.430,0.781,P<0.05 or P<0.01);the content of HSAY was negatively associated with storage life (r=-0.777,P<0.01). There was no correlation between the remaining variables (P>0.05). CONCLUSIONS :The longer the storage life ,the darker the color and the lower the content of HSYA ,so it is not suitable for over year and multiyear preservation.