Anti-inflammatory Activity Of Lonicera Fulvotomentosa

Lonicera fulvotomentosa,belong to genus Lonicera,is one of the resoure plants of Shanyinhua,a Chinese Traditional Medicine collected in the Chinese Pharmacopoeia.It is also used as an ecological improving plant for the controlling karst rocky desertification in Guizhou Province,resulting in the huge biomass of the plant.The flowers and buds of Lonicera fulvotomentosa are used as a substitute of honeysuckle for many years,for the treatment of heat toxic blood stasis,wind-heat flu,etc.Its chemical constituents consist of phenolic acid and triterpenoids saponins,and many biological activities are found based on the chemical constituents,such as anti-inflammatory,anti-viral,anti-oxidation,liver-protecting and anti-tumor activities,while the anti-inflammatory is the main pharmacological activity for the traditional use of the herb.Inflammation is recognised to play fundamental role in the pathogenesis of many diseases,and complement system has always activated during the inflammation reaction,and the products of the complement activation cascades were considered as the leading to endothial cell injury.In this paper,the CVF,a specific activator for the complement alternative pathway,was used to induce acute lung injury?ALI?in mice,and the effects of the extact of Lonicera fulvotomentosa against the inflammatory injury were evaluated.Firstly,mice were randomly divided into control,model,three part extracts(500mg·kg^-1,300 mg·kg^-1,100 mg·kg^-1)doses groups,the mice in control group and model group were intragastrically administered with 0.2%CMCNa.After 7 days,35?g·kg^-1CVF was injected to mice to induce ALI,and then the alveolar lavage fluid?BALF?was collected to record the number of cells,and the activity of myeloperoxidase?MPO?was detected in the lung lobe.The screening results showed that several extracts of Lonicera fulvotomentosa could decrease the number of cells in BALF and MPO activity,and improved the ALI inflammatory response induced by activation of the complement alternative pathway.Therefore,TGF which contains n-butanol and ethyl acetate was selected to an active site.Scecondly,we investigated the effects of TGF(500 mg·kg^-1,100 mg·kg^-1)groups on CVF-induced ALI in mice.We detected the expression of IL-6,TNF-?,P-selectin and ICAM-1 in BALF and serum;The pathological condition of the lung lobe was observed by H&E staining with the lung lobe fixed in 4%paraformaldehyde.The expression of complement components C5a,C5b-9 and the degree of phosphorylation of NF-?B p65 and JAK2 protein were detected by immunohistochemistry.At the same time,the serum of each group was collected before and after oral administration to detect complement activity.The results showed that compared with the model group,TGF and PDTC groups could decrease the number of cells in BALF and the MPO activity,down-regulated the expression of inflammatory factors ICAM-1,P-selectin,IL-6 and TNF-?in BALF and serum,and improved the inflammatory infiltration of lung tissue in mice,also down-regulated the expression of complement factor C5a and C5b-9 in lung tissue.The results of TGF and PDTC groups improved the phosphorylation of NF-?B p65 and JAK2 proteins suggested that its mechanism may be related to the inhibition of NF-?B and JAK2 signaling pathway activation,but TGF itself does not affect serum complement activity,which indicated that TGF could not inhibit complementation but may act by inhibiting the inflammatory response produced by complement activation.Finally,the effect of TGF injury of HMEC induced byH₂O₂ was conducted,directly from the effects on the cell morphology and viability.Furthermore,experiment the expression of NF-?B p65 protein and its phosphorylation degree were determined with methods of Western blot,as well as the related apoptosis proteins like Bax Bcl-2,and caspase-3.The results show that 100?g·mL^-11 dose of TGF can reduce the mortality of cells injured by H₂O₂,down-regulate the expression of NF-?B p65 protein,and reduce the ratio of Bax/Bcl-2.Its indicated that TGF could inhibit apoptosis of HMEC,and its mechanism may be related to inhibition of NF-?B signaling pathway activation and the expression of related apoptotic proteins,but the activation of caspase-3 protein is not detected in the experiment,which may be related to the experimental time.In summary,the total glycosides were the anti-inflammatory active fraction of Lonicera fulvotomentosa,improving the acute lung injury induced by CVF and reducing the inflammatory response in mice.The mechanism may be related to NF-?B and JAK2 signaling pathway,and the active constituents of Lonicera fulvotomentosa may be involved in the regulation of oxidative stress-induced injury process.