Black currant, Ribes nigrum L. was analysed as a trial in developing natural material.
Black currant berries were extracted by the methods of hot water distillation(BCD) and ethanol extraction(BCE). The antioxidative activity of BCD and BCE was accessed through total polyphenol and flavonoid co...
Black currant, Ribes nigrum L. was analysed as a trial in developing natural material.
Black currant berries were extracted by the methods of hot water distillation(BCD) and ethanol extraction(BCE). The antioxidative activity of BCD and BCE was accessed through total polyphenol and flavonoid contents, DPPH and ABTS radical scavenging activities, SOD activity and ROS production rate in LPS treated Raw 264.7 cells. In order to measure anti-inflammatory effect of BCD and BCE, productions of NO and pro-inflammatory cytokines were measured and mRNA expression level of the pro-inflammatory cytokines were analysed in LPS treated Raw 264.7 cell. In addition, on the 7th week of the experiment, cytokine production and immunocyte quantity were measured in serum and tissue examination was performed through H&E stain within the tissue of inflammation-induced Balb/c mouse.
Total polyphenol content of BCD and BCE was represented as 202.1±1.0 ㎎/g and 238.3±2.6 ㎎/g, respectively. DPPH, ABTS radical scavenging activity of BCD and BCE increased concentration-dependently and this phenomenon was revealed more in BCD. Both at all concentrations showed no cytotoxicity and suppressed ROS production in Raw 264.7 cells. In Raw 264.7 cells, SOD activity of BCE was more increased in concentration-dependency than that of BCD. NO production was decreased by BCD and BCE and production of IL-1β, IL-6 and TNF-α was suppressed by BCD and BCE at all concentrations as compared with the control group significantly. mRNA expession level of IL-1β, IL-6 and TNF-α was decreased at 100 ㎍ / ㎖, at 10, 100 and at all concentrations of BCE, respectively.
In Balb/c mouse serum, BCD- and BCE-administered groups suppressed production of IL-1β, IL-6, TNF-α and IFN-γ at all the concentrations significantly in comparison to the control group. The Leukocyte number was decreased but the eosinophil, neutrophil and basophil increased and the lymphocyte slightly increased in number at all the BCD- and BCE-administered groups in comparison to control group. On the other hand, number of monocyte was significantly decreased in all the BCD- and BCE-administered groups.
It was observed that there was no difference of epidermis thickness between control group and experimental group of BCD and BCE on the back skin of the LPS treated Balb/c mouse as a result of H&E staining but the lymphocyte number of dermis in BCD and BCE group was counted less than the control group. It was confirmed by naked eye and visible light microscope at magnification of 40× that a thickness of ear tissue for control group was thickened while that of BCD and BCE group was thinned. In the liver tissue there was no change in shape of nucleus for all the participants after the 7th week of the experiment while the Kupffer's cell number of BCD and BCE group was fewer than that of the control group.
From the above results it was confirmed that the BCD and BCE have anti-oxidant activity and anti-inflammatory effect in Raw 264.7 cell and Balb/c mouse model induced with LPS. It is considered that the BCD and BCE have a potential for developing immune medicinal materials for preventing inflammatory edema and cellular damage and an activated substance of natural material relevant to the cosmetics industry and skin disease. It is expected that further research on BCD and BCE should be carried out through systematic and clinical studies and this will be dedicated to development of the functional cosmetics and medicine material with common antioxidant, anti-inflammation effect and widely used for maintaining and improving skin condition and health.
Keywords : Ribes nigrum L., antioxidation, anti-inflammation, ROS, Cytokine
Black currant, Ribes nigrum L. was analysed as a trial in developing natural material.
Black currant berries were extracted by the methods of hot water distillation(BCD) and ethanol extraction(BCE). The antioxidative activity of BCD and BCE was accessed through total polyphenol and flavonoid contents, DPPH and ABTS radical scavenging activities, SOD activity and ROS production rate in LPS treated Raw 264.7 cells. In order to measure anti-inflammatory effect of BCD and BCE, productions of NO and pro-inflammatory cytokines were measured and mRNA expression level of the pro-inflammatory cytokines were analysed in LPS treated Raw 264.7 cell. In addition, on the 7th week of the experiment, cytokine production and immunocyte quantity were measured in serum and tissue examination was performed through H&E stain within the tissue of inflammation-induced Balb/c mouse.
Total polyphenol content of BCD and BCE was represented as 202.1±1.0 ㎎/g and 238.3±2.6 ㎎/g, respectively. DPPH, ABTS radical scavenging activity of BCD and BCE increased concentration-dependently and this phenomenon was revealed more in BCD. Both at all concentrations showed no cytotoxicity and suppressed ROS production in Raw 264.7 cells. In Raw 264.7 cells, SOD activity of BCE was more increased in concentration-dependency than that of BCD. NO production was decreased by BCD and BCE and production of IL-1β, IL-6 and TNF-α was suppressed by BCD and BCE at all concentrations as compared with the control group significantly. mRNA expession level of IL-1β, IL-6 and TNF-α was decreased at 100 ㎍ / ㎖, at 10, 100 and at all concentrations of BCE, respectively.
In Balb/c mouse serum, BCD- and BCE-administered groups suppressed production of IL-1β, IL-6, TNF-α and IFN-γ at all the concentrations significantly in comparison to the control group. The Leukocyte number was decreased but the eosinophil, neutrophil and basophil increased and the lymphocyte slightly increased in number at all the BCD- and BCE-administered groups in comparison to control group. On the other hand, number of monocyte was significantly decreased in all the BCD- and BCE-administered groups.
It was observed that there was no difference of epidermis thickness between control group and experimental group of BCD and BCE on the back skin of the LPS treated Balb/c mouse as a result of H&E staining but the lymphocyte number of dermis in BCD and BCE group was counted less than the control group. It was confirmed by naked eye and visible light microscope at magnification of 40× that a thickness of ear tissue for control group was thickened while that of BCD and BCE group was thinned. In the liver tissue there was no change in shape of nucleus for all the participants after the 7th week of the experiment while the Kupffer's cell number of BCD and BCE group was fewer than that of the control group.
From the above results it was confirmed that the BCD and BCE have anti-oxidant activity and anti-inflammatory effect in Raw 264.7 cell and Balb/c mouse model induced with LPS. It is considered that the BCD and BCE have a potential for developing immune medicinal materials for preventing inflammatory edema and cellular damage and an activated substance of natural material relevant to the cosmetics industry and skin disease. It is expected that further research on BCD and BCE should be carried out through systematic and clinical studies and this will be dedicated to development of the functional cosmetics and medicine material with common antioxidant, anti-inflammation effect and widely used for maintaining and improving skin condition and health.
Keywords : Ribes nigrum L., antioxidation, anti-inflammation, ROS, Cytokine
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