Studies On Anti-Inflammation Effects And Molecular Mechanism Of Chlorogenic Acid Extracted From Lonicera Hypoglauca Miq

Objective To provide a theory and technical support to further develop of the medicinal plants of Guangxi, accelerated the progress of modernization of traditional chinese medicine, we observed the anti-inflammatory effects of Guangxi Lonicera hypoglauca Miq. and its acute toxicity. In the same time, investigated the molecular mechanism of anti-inflammatory of chlorogenic acid,extracted from Lonicera hypoglauca Miq. in-vitro.Methods①Desiccative Lonicera hypoglauca Miq. were extracted by ethanol, isolated and purificated it with macroporous resin. After concentrating and drying, High performance liquid chromatography (HPLC) was used for evaluating extracts.②Divided randomly into model group, positive control group, low-dose group, middle-dose group and high-dose group. Dosage of administration was respectively 20,15,10g·kg-1.Croton oil-induced ear swelling, carrageenan-induced hind paw edema were prepared to study the effects of Lonicera hypoglauca Miq. on inflammation. And acute toxicity experiment was done.③Collected, purified, cultivated peritoneal macrophages (PMФ) of a rat , randomly divided into blank control group, LPS group and calcium ionophore (A23187) group, positive control group and chlorogenic acid groupsⅠ—Ⅲ, dosage of administration was respectively 50,100,200 mg·L-1. Inflammation model of cytes were prepared with LPS and A23187 , the change of cell morphology was observed under the convert microscope. MTT assay was used to detect the effects on cells growth activities of chlorogenic acid. PMФwas stimulated with LPS for a prolonged period , ELISA was used to detect the level of TNF-α, IL-6 and PGE2 in the supernatant, COX-2 activity was determined by the level of PGE2 in the supernatant. After stimulating PMФwith A23187 for a short time, the 6-keto-PGF1αlevel in the supernatant was measured by radioimmunoassay, in order to express COX-1 activity.Results①The content of chlorogenic acid was 24.3%.②The ear edema induced by croton oil was inhibited significantly by each dose, high dose group had no difference compare with positive control group.Carrageenan induced hind paw edema was also inhibited significantly by middle-dose group and high-dose group .All mice were survival in a maximum dose.③Most of the cell morphology were changed from round to fusiform after stimulated by LPS.④Chlorogenic acid had no inhibitive effects between31.25 mg·L-1 and 1000 mg·L-1.⑤In normal groups, TNF-α, IL-6 and PGE2 were only a little expression while strongly expressed in LPS-induced groups. The level of TNF-α, IL-6 and PGE2 in drug groups were lower than that in LPS-induced group, and the difference were significant (P<0.01) . The concentration of 50mg·L-1 group was ineffective in the expression of TNF-α. In a dose-dependent manner.⑥6-keto-PGFlαstrongly expressed after induced by A23187. Different Chlorogenic acid concentrations had an inhibitory or promoting effect on the growth of 6-keto-PGFlα.Conclusions Guangxi Lonicera hypoglauca Miq. has a significant acute anti-inflammatory effect and low toxicity. The anti-inflammatory effect of chlorogenic acid may be related to inhibiting TNF-α,IL-6 activity and affecting activity of COX-2 of exogenous AA metabolism.