The Study On Main Chemical Components Of Total Flavonoids Of Hyssopus Officinalis L. And Activities Of Antioxidant,Anti-inflammatory

Objective:To investigate the main chemical constituents of total flavonoids of Hyssopus officinalis L and its anti-oxidation and anti-inflammatory effects in vitro,and to provide experimental basis for its further development and utilization.Methods:1)Qualitative analysis of the main compounds of total flavonoids of Hyssopus officinalis L by high performance liquid chromatography（HPLC）;2)The anti-oxidative activity of total flavonoids and ethanol extract of Radix Angelicae Sinensis in vitro were evaluated by DPPH free radical scavenging ability and hydroxyl radical scavenging ability assay;3)The LPS-induced RAW264.7 cell inflammation model was based on the intervention of total flavonoids of Hyssopus officinalis L,IL-6,TNF-α,MCP-1,PGE2,NO and IL-Quantitative PCR（q-PCR）was used to detect the mRNA expression of TNF-α,IL-6 and MPC-1.（4）The nuclear factor NF-κB was used as an index to establish a NF-κB dual luciferase reporter assay system,and to explore the inhibitory effect of total flavonoids of Hyssopus officinalis L and its main components on NF-κB.Results:1)The main compounds of total flavonoids of Hyssopus officinalis L may contain flavonoids such as luteolin,acacetin,rutin,diosmetin,apigenin and rosmarinic acid,caffeic acid,Acid,ferulic acid and other phenolic acids.The content of rosmarinic acid was 6.404mg/g and the content of chlorogenic acid was 2.286mg/g;2)Total flavonoids and different concentrations of ethanol extracted from Radix Shenmu had the ability of scavenging DPPH·free radical,OH·radical and O₂^-radical scavenging ability and reducing ability,among them,the total flavonoids had significant antioxidant capacity,followed by70%Ethanol extract,more than 50%,30%and water extract,but lower than the positive control drug Vc antioxidant activity;3)The total flavonoids of Hyssopus officinalis L had no effect on the expression of LDH in RAW264.7 cells.There was no significant difference between the two groups（P>0.05）.It can inhibit the expression of PGE2,IL-6,MCP-1,NO,TNF-αand IL-6,MCP-1 and TNF-αThe expression level was dose-dependent;4)total flavonoids of Hyssopus officinalis L and rosmarinic acid could inhibit the activity of NF-κB,the difference was statistically significant（P<0.05）.Among them,the high concentration total flavonoids of Hyssopus officinalis L and rosmarinic acid inhibited NF-κB activity better than DEX.The inhibitory activity of NF-κB in high concentration total flavonoids of Hyssopus officinalis L was equivalent to that of high concentration rosmarinic acid,with no significant difference（P>0.05）.Conclusion:total flavonoids of Hyssopus officinalis L may contain flavonoids and phenolic acids,of which rosmarinic acid and chlorogenic acid are higher.Its anti-oxidation,anti-inflammatory activity was significant.Rosmarinic acid may be the main active ingredient that inhibits the activity of NF-κB.