Extraction of chlorophyll from Forsythia suspensa leaves using cloud point extraction coupled with ultrasonication was investigated in the present study.Single factor test and orthogonal design were used to determine the optimal conditions with absorbance values of 665 nm as the index of evaluation.The optimal conditions for the extraction of chlorophyll were as follows:Triton X-114 concentration of 4%,ultrasonic power of 250 W,ultrasonic time of 11 min,and solid-liquid ratio of 1:10.Comparing the developed extraction method with the traditional organic solvent extraction methods and ultrasonic extraction method,the extraction rate improved 9.92% and 11.58%,respectively.This study provided a new method for the extraction of chlorophyll with the following advantages:low temperature (room temperature) of extraction,no using volatile or toxic organic solvents,economical,safe,efficient,convenient,time-saving,environmental protection,etc.In addition,the method can be applied for the extraction of chlorophyll from other natural resources.

The objective of this study was to determine the content of total lignans,forsythiaside A,forsythiaside B and phillyrin in Forsythia suspensa leaves from Henan province and Shanxi province.The total lignans of F.suspensa leaves from Shanxi province were higher than Henan province by spectrophotometry method.The contents of forsythiaside A,forsythiaside B and phillyrin were determined by HPLC.The average recoveries of forsythiaside A,forsythiaside B and phillyrin were 99.6% (RSD=1.23%),101.3% (RSD=1.74%) and 102.3% (RSD=2.58%),respectively.The experimental results showed that the content of forsythiaside A and phillyrin in F.suspensa leaves from Shanxi and Henan was relatively high.In addition,the content of phillyrin in F.suspense leaves from different habitats were of large difference. F.suspensa leaves are rich in forsythiaside and lignans,more researches on F.suspensa leaves are still needed to confirm whether it can be used as substitute or cooperate with F.suspensa fruit for medicinal application.

To investigate the anti-inflammatory activities of Forsythia suspensa extract (FSE) with emphasis on excretion of inflammatory cytokines by LPS-induced BEAS-2B inflammation model.BEAS-2B cell viability was detected by MTT method in order to screen out the optimal drug dose of F.suspensa ethanol extracts (FSEE),60% forsythiaside A (60% FTA) extracts and 90% forsythiaside A (90% FTA) extracts;The morphology of BEAS-2B cells treated with FSE were observed by microscope; NO content of cell culture supernatant was determined by Griess Reagent method;Intracellular reactive oxygen species (ROS) was analyzed by flow cytometry.Toxical experimental results revealed that FSE did not show toxic reaction to BEAS-2B cell at concentration of less than 128 μg/mL;while both 60% FTA and 90% FTA showed toxicity at the concentration between 128 and 256 μg/mL.Anti-inflammatory results showed that all FSE can inhibit the secretion of NO (P<0.01) at concentration 25,50,and 100 μg/mL,and 90% FTA extracts exhibited a dose-dependent manner;The elevated level of intracellular ROS in LPSstimulated BEAS-2B cells was significantly diminished by all FSE in a dose-effect manner (P<0.05).In addition,the therapeutic effect of FSEE,60% FTA and 90% FTA increased gradually.It was concluded that the 90% FTA extracts were the main active fraction of anti-inflammatory effect and forsythiaside A was possibly the active ingredient.The results presented in this study will be helpful for the further study of F.suspensa on anti-inflammatory activity.

In this study,the anti-inflammatory effect of major components of Forsythia suspensa was explored and its bioactive components with anti-inflammatory effect were screened to provide reference for the development of new anti-inflammatory drugs of F.suspensa. Then xylene-induced mice ear swelling models were established and the swelling degree was calculated.The content of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in mice serum were detected by enzyme-linked immunosorbent assay(ELISA).The swelling degree and the content of TNF-α and IL-6 in mice serum of medication administration groups were compared with those of control group respectively.Compared with the control group,phillyrin had no significant inhibition effect on swelling degree or the production of TNF-α and IL-6,and there was no significant difference between the two groups(P>0.05).Compared with the control group,significant differences of the swelling degree and the production of TNF-α and IL-6 were detected in phillygenin,forsythiaside A and forsythiaside B group respectively(P<0.05).Thus it can be seen that by oral administration,only the three selected components of F.suspense,including phillygenin,forsythiaside A and forsythiaside B,had anti-inflammatory effects.By oral administration,the anti-inflammatory effect of phillyrin had not been found yet,but phillygenin,forsythiaside A and forsythiaside B had stronger anti-inflammatory effects.Meanwhile,it can be speculated that the anti-inflammatory mechanism of F.suspensa may be related with its inhibition effect of the production of TNF-α and IL-6 and the effect of multiple components.

A spectrum effect correlation pattern was developed to evaluate the antioxidant activity and the main active components of Forsythia suspense leaves in this study.The fingerprint chromatogram of the 18 Forsythia suspense leaves extracts collected from different places was established by HPLC.In addition the antioxidant activity and the content of total phenol of these 18 extracts were studied using the 1,1-diphenyl-2-picrylhydrazyl free radical(DPPH·)-scavenging method and Folin-Ciocalteuas method,respectively.The results showed that the antioxidant activity was correlated to the content of total phenol by the analysis of bivariate correlation.Furthermore,three components played a role in antioxidant activity of F.suspense leaves through the analysis of principal component,bivariate correlation and multiple linear regression.The study provided reference for the evaluation of antioxidant activity of F.suspense leaves.

Antipyretic effect and mechanisms of  Forsythia suspensa extract and forsythia oil on rats with high fever induced by yeast were investigated in the study. Rat model of yeast-induced fever was established by subcutaneous injections of yeast. After the basal body temperature was measured, the rats were randomly divided into four groups:control group,model group, Forsythia extract group (25 crude drug g/kg) andforsythia oil group (26.7 crude g/kg). Then the body temperature and the concentration of prostaglandin E2 (PGE-2) and cyclic adenosine monophosphate (cAMP) in the hypothalamus cerebrospinal fluid of rats were detected dynamically two hours after dosing. The result showed that  F.suspensa extract and forsythia oilcan significantly decrease the temperature(P<0.01 or  P <0.05)as well as hypothalamus cAMP content (P<0.01) in yeast-induced fever rats.In addition, F.suspensa  extract also obviously decreased hypothalamus PGE-2 content (P<0.01). F.suspensa extract can down-regulate cAMP and PGE-2 in hypothalamus. And forsythia oil down-regulate cAMP in hypothalamus. Both can be used as symptomatic treatment for clinical fever.

This is the first time to investigate the chemical constitutes of Forsythia suspensa flower for tyrosinase-inhibition activity. First, different organs (flower,leaf,green Forsythiae Fructus and ripe Forsythiae Fructus) of F.suspensa  were tested for the tyrosinase-inhibition activity,and the F.suspensa flower showed concentration-dependent on the inhibition of tyrosinase activity.Second,different extracts of F.suspensa flower were also tested for the tyrosinase-inhibition activity,and the 95% ethanol extract displayed the best inhibition of tyrosinase activity.Then,five compounds were obtained from 95% ethanol extract by the activity-guided isolation method,and their structures were identified as forsythiaside A (1),rutin (2),(+)-pinoresinol-β-D-glucoside (3),(+)-pinoresinol monomethyl ether β-D-glucoside (4) and (+)-phillyrin (5),while compound 4was reported for the first time from F.suspensa flower.Four of them showed weak inhibition activity against tyrosinase but displayed obvious synergistic effect.The results implied that the 5 compounds were most likely to be the chemical substance foundation of the whitening effect of F.suspensa flower and its inhibitory effect was possibly produced by synergistic effect.

To investigate the effects of volatile oil of Forsythia suspense (VOFF) on different rat models,and explore the mechanism of its anti-inflammatory and antipyretic effects.Two rat fever models were established by subcutaneously injection of dry yeast and intraperitoneal injection of lipopolysaccharide (LPS) to evaluate the antipyretic effect of VOFF,respectively.Rats cotton ball granuloma model,rats foot swelling model induced by subcutaneous injection of carrageen or fresh egg white at the right posterior foot were used to evaluate the anti-inflammatory effects.VOFF can significantly inhibit LPS and dry yeast induced fever,reduce carrageenan and fresh egg white induced rat paw edema,compared with the model group (P<0.05).It can also significantly inhibit the proliferation of granulation tissue,compared with the model group (P<0.01).The results showed that VOFF had obvious antipyretic and anti-inflammatory effects,which provided a theoretical basis for further research on heat clearing and detoxifying function.

To obtain endophytes which can produce antimicrobial substance from forsythia,endophytic fungi strains were isolated and purified from fresh tissues of forsythia,morphology analysis was used and antimicrobial activitywas detected by oxford cup method,and 18S rRNA gene sequence was analyzed.The forsythin,forsythia glycosides of A and forsythia glycosides of B were detected in their fermented liquid by HPLC.The results showed that five endophytic fungi strains were obtained.Four of them had good antimicrobial activity on S.Aureus and S.enteritidis.Molecular analysis and NCBI blast showed that three strains were Cladosporium and one strain was Aspergillus flavus.The HPLC detection suggested that certain concentration of forsythin,forsythia glycosides of A and forsythia glycosides of B were existed in their supernatant of fermented liquid,but further research is need to reveal the relation between the forsythin,forsythia glycosides of A and forsythia glycosides of B and the antimicrobial activity.This study provided a new way and new sources of strains for produce antibacterial material by microbial fermentation.

The volatile oil from Forsythia suspense possesses antibacterial,anti-oxidant,antipyretic and anti-inflammatory activities.It is the major fraction responsible for the pharmacological effect of this plant.It has potential application value in the food industry.This review summarized the chemicals in the volatile oils from different parts of F.suspense and in compatibility drugs,and the pharmacological effects of these oils in order to provide reference for the deep development and comprehensive utilization of volatile oil from Forsythia suspense.

To study the anti-inflammatory effect and anti-inflammatory mechanism of Phillygenin,dexamethasone as a positive control,established a lipopolysaccharide (LPS)-induced RAW264.7 inflammation model,to detect the release of inflammatory factors and the expression of related proteins and mRNA,in order to improve the comprehensive understanding of the anti-inflammatory effect of Phillygenin and provide a strong scientific basis for the clinical development of Phillygenin.The NO production was determined by assaying nitrite in culture supernatants with the Griess reagent.The levels of TNF-α and IL-6 in culture media were measured with ELISA kits.Western blot assay was performed to illustrate the inhibitory effects of Phillygenin on iNOS and COX-2.RT-qPCR was detected for mRNA expression of iNOS and COX-2.Treatment with Phillygenin or Dexamethasone dose-dependently inhibited the production expression of NO,TNF-α and IL-6 in RAW264.7 macrophages.Western blot and RT-qPCR assay suggested that the mechanism of the anti-inflammatory effect was associated with the inhibition of iNOS and COX-2.In conclusion,Phillygenin exhibited obvious anti-inflammatory effect and its mechanism may be related to its regulation on inflammatory cytokines,and inhibition of the expression of iNOS and COX-2.

The content of main active components in Forsythia suspensa leaves and flowers from different regions were determined,and the quality difference in samples was analyzed,which provided a scientific basis for the resource development and comprehensive utilization of F. suspensa leaves and flowers.The contents including rutin,hyperoside,forsythiaside A,forsythin,oleanolic acid and ursolic acid in 28 batches of F. suspensa  leaves and flowers were determined by liquid chromatography mass spectrometry (LC-MS) technology,the results were analyzed by principal component analysis (PCA).The results showed there were some differences in the main active components in F. suspensa  leaves and flowers from different regions.PCA showed the comprehensive evaluation of F. suspensa  leaves in Shanxi Linfen,Shaanxi Xi'an and Henan Xinyang were better,and the comprehensive evaluation of F. suspensa  flowers in Henan Luanchuan,Shandong Pingyi and Shandong Rizhao were better.The method of good precision,repeatability and stability showed certain reference to the comprehensive utilization and the development of resource products of F. suspensa  leaves and F. suspensa flowers.