Chemical Constituents And In Vitro Pharmacological Activities Of The Flower Of Zingiber Striolatum Diels.

Zingiber striolatum Diels.is a medicinal and edible plant.Its flower is widely used as a vegetable to treat diabetes and constipation.At present,there are few studies on the chemical constituents and pharmacological activities of the flower.Therefore,in this research,the chemical constituents and in vitro pharmacological activities of the water extract（WE）and 70%ethanol extract（EE）were studied.It provides a theoretical basis for the development and utilization of Z.striolatum.The main work contents of this paper are as follows:The contents of total phenolic acids and total flavonoids in the WE and EE were determined by UV spectrophotometer.The results showed that both the WE and EE contained higher contents of total phenolic acids and total flavonoids.The contents of total phenolic acids and total flavonoids in the EE were significantly higher than those in the WE.The gallic acid equivalent and rutin equivalent of the EE were 36.01±1.04 mg GAEs/g extract and 58.15±1.14 mg REs/g extract,respectively.UPLC-Q-Orbitriap MS was used to identify the chemical constituents of the WE and EE from flower of Z.striolatum.A total of 44 chemical constituents were identified,35 compounds in the WE and 24 compounds in the EE were identified,including 6 phenolic acids in the WE and EE:which were gallic acid,vanillic acid,p-Hydroxy-cinnamic acid,homovanillic acid,6-gingerol and metilox.including 14flavonoids in the WE and EE:which were D-（+）-catechin,epicatechin,hesperetin,morin,isorhamnetin,rhoifolin,isoquercitrin,procyanidin B2,vicenin II,narcissoside,miquelianin,typhaneoside,rutin and diosmin.DPPH and ABTS method were used to study the antioxidant effect of the WE and EE from flower of Z.striolatum.The results showed that the WE and EE had a certain scavenging effect on DPPH and ABTS free radicals.Among them,the antioxidant activity of the EE was significantly stronger than that of the WE.The ascorbic acid equivalent of DPPH free radical scavenging rate of the EE was 38.63±3.98 mg AEs/g extract,and the ascorbic acid equivalent of ABTS free radical scavenging rate was 52.22±4.05 mg AEs/g extract.The paper diffusion method was used to study the antibacterial effect of the WE and EE from flower of Z.striolatum.The results showed that the WE and EE had certain inhibitory effect on the six strains.Among them,the WE and EE had better inhibition effect on Proteus vulgaris,with inhibition zone size of 17.62±0.09 mm and 11.86±0.31 mm,respectively.The WE had better inhibition effect on Staphylococcus aureus,with inhibition zone size of 10.75±0.18 mm.The inhibitory activities of the WE and EE on tyrosinase,α-glucosidase,acetylcholinesterase and butyrylcholinesterase were determined by multi-function microplate.The results showed that the WE and EE had certain inhibitory effect on the four enzymes.The IC₅₀ of the WE and EE were 7.84±1.32μg/m L and 22.91±4.46μg/m L,respectively.The acarbose equivalent was 36.92±6.74 mmo L ACEs/g extract and 12.67±2.33 mmo L ACEs/g extract,respectively.The inhibitory effect of the WE and EE onα-glucosidase was significantly stronger than acarbose.Mouse monocyte macrophage RAW264.7 cells was used as the research object to induce inflammatory response by LPS.The effects of the WE and EE from flower of Z.striolatum on the inflammatory response of RAW264.7 cells were studied with the content of NO and the concentrations of TNF-αand IL-6 as indexes.The results showed that both the WE and EE could inhibit the release of NO,TNF-αand IL-6 in LPS induced RAW264.7 cells in a concentration dependent manner,the higher the concentration,the stronger the inhibitory effect.Because the inhibitory activity of the EE was stronger than that of the WE,so the EE was selected for follow-up study.The results of morphological observation showed that the EE could inhibit LPS induced inflammatory reaction in RAW264.7 cells,and the inhibitory effect was concentration dependent,the higher the concentration was,the stronger the inhibitory effect was.The level of ROS in each group was detected by the fluorescence intensity of ROS.The results showed that the EE reduced the level of ROS in RAW264.7 cells induced by LPS in a concentration dependent manner.The m RNA expressions of IL-6,TNF-α,i NOS,IL-1βand COX-2 were detected by real time PCR.The results showed that the EE could down regulate the expression of i NOS,COX-2,IL-1β,TNF-αand IL-6 m RNA in RAW264.7 cells induced by LPS,and the high concentration（128μg/m L）group had significant inhibitory effect.Western blot and immunofluorescence analysis showed that the EE high concentration group could significantly down regulate the expression of i NOS and COX-2 protein in RAW264.7 cells induced by LPS,the EE inhibited the phosphorylation of p38,IκBαand NF-κB p65,inhibited the transfer of NF-κB from cytoplasm to nuclear in RAW264.7 cells induced by LPS,and thus inhibited the i NOS,COX-2,IL-1β,TNF-αand IL-6 m RNA transcription in RAW264.7 cells induced by LPS,inhibited the production of inflammatory mediators.All of the above results showed that EE could repress the activation of p38 MAPK and NF-κB signaling pathways.In conclusion,the WE and EE from flower of Z.striolatum have higher contents of total phenolic acids and total flavonoids,strong antioxidant activity,α-glucosidase inhibitory activity and anti-inflammatory activity.The EE could reduce the production of ROS in RAW264.7 cells induced by LPS,could down regulate the expression of i NOS,COX-2,IL-1β,TNF-αand IL-6 m RNA in RAW264.7 cells induced by LPS,and the EE high concentration group could significantly down regulate the expression of i NOS and COX-2 protein in RAW264.7 cells induced by LPS,the EE inhibited the phosphorylation of p38,IκBαand NF-κB p65,inhibited the transfer of NF-κB from cytoplasm to nuclear in RAW264.7 cells induced by LPS,and thus inhibited the i NOS,COX-2,IL-1β,TNF-αand IL-6 m RNA transcription in RAW264.7 cells induced by LPS,inhibited the production of inflammatory mediators.The EE repressed the activation of p38 MAPK and NF-κB signaling pathways in RAW264.7 cells,and then regulated the occurrence and development of inflammatory response.