Studies On The Chemical Composition Of Essential Oil From Ginger And The Anti-inflammatory Mechanisms Of Its Main Active Component 6-Gingerol

Objective: Ginger is the fresh rhizome of the perennial monocotyledonous ginger of the genus Zingiberaceae.It is the most common medicine and food homologous traditional Chinese medicine.Ginger volatile oil and its processed products studied constituents,and to investigate the possible reasons for the differences in the pharmacological effects;ginger different origin and the content of 6-gingerol ginger 6-direct pressing of gingerol extraction rate detector for correlation ginger product development and materials provide reference and new ideas;predict the potential anti-inflammatory target of 6-gingerol by MOE and other software,and verify the anti-inflammatory targets through molecular biology related techniques to explore the main ginger organisms.The anti-inflammatory mechanism of the active ingredient 6-gingerol.Provide theoretical basis for target selection and research and development of ginger-related drugs.Methods: The volatile oils of ginger,dried ginger,artillery ginger and ginger charcoal were extracted by steam distillation.The volatile oils were diluted by ether and analyzed by gas chromatography-mass spectrometry(GC-MS)and then compared the results.6-gingerol in Ginger from five different habitats was extracted by alcohol reflux extraction method.Ginger juice and ginger dregs were obtained by direct pressing method,and 6-gingerol in fresh ginger and ginger dregs was extracted by alcohol reflux method.The content of 6-gingerol in each sample was determined by high performance liquid chromatography and compared.The potential anti-inflammatory targets of 6-gingerol were explored by MOE software,and the anti-inflammatory mechanism of 6-gingerol was studied on this basis.MTT method was used to determine the concentration of 6-gingerol,ELISA method was used to detect the effect of 6-gingerol on the secretion of inflammatory factors TNF-?,IL-6and c AMP,and Western Blot method was used to detect the effect of 6-gingerol on the expression of phosphodiesterase 4 in inflammatory cells.Results: A total of 48 compounds were identified from the volatile oils of four different processed products of ginger,including 30 volatile oils of ginger,36 volatile oils of dried ginger,35 volatile oils of artillery ginger and 26 volatile oils of ginger charcoal.Among them,ginger contains more beta-hydrocelene,citral and camphene;dried ginger mainly contains gingerene,beta-sesquiterpene,beta-hydrocelene,etc.;artillery ginger mainly contains gingerene,beta-hydrocelene and so on;ginger charcoal mainly contains borneol,gingerene,beta-hydrocelene and so on.The content of 6-gingerol in Ginger from different producing areas was 0.123 mg/m L in Liaoning,0.121 mg/m L in Shandong,0.176 mg/m L in Yunnan,0.274 mg/m L in Sichuan and 0.264 mg/m L in Jiangxi.The content of 6-gingerol in 95.57 g ginger was 121.9 mg and that in 66 m L ginger juice was 55.39 mg,which was 45.46% of the total content of6-gingerol in ginger.The content of 6-gingerol in 4.98 g ginger residue was 36.68 mg,which was the total content of gingerol in ginger.The content was 30.10%.PDE4 is a possible anti-inflammatory target of 6-gingerol screened by Pharm Mapper.The docking of PDE4 and 6-gingerol by MOE simulation showed that the target had a good docking with 6-gingerol.Compared with the blank group,the concentration of6-gingerol in 25-100 ?g/m L group was P > 0.05,indicating that 6-gingerol had no effect on the viability of RAW264.7 cells;LPS could activate RAW264.7 and significantly increase its viability.The activity of RAW264.7 cells induced by LPS was significantly inhibited in the 6-gingerol administration group compared with the LPS control group(P< 0.05 or P < 0.01).Compared with the LPS control group,the secretion of TNF-? and IL-6 in the 6-gingerol administration group was significantly decreased,and the inhibition of TNF-? and IL-6 secretion was positively correlated with the concentration of administration.The secretion of c AMP in the administration group was significantly higher than that in the LPS control group.The expression of PDE4 protein in RAW264.7cells induced by LPS was significantly higher than that in the blank group(P < 0.05).The expression of PDE4 protein in RAW264.7 cells treated with LPS was significantly higher than that in the control group(P < 0.05 or P < 0.01).The expression of PDE4 protein in RAW264.7 cells induced by LPS was significantly inhibited by 6-gingerol at 25-100?g/m L concentration compared with that in LPS group(P < 0.05 or P < 0.01).The degree of inhibition was positively correlated with the concentration of the drug.Conclusion: The volatile oils of the four processed ginger products have the same composition and their own characteristics.The kinds and contents of compounds in volatile oil of ginger vary in different processing processes,and the changes in components may lead to differences in pharmacological effects of processed products.Among the 6-gingerol contents detected by high performance liquid chromatography in Ginger from different producing areas,the concentration of 6-gingerol in Sichuan and Jiangxi provinces is significantly higher than that in other three producing areas.The content of 6-gingerol in ginger juice obtained by direct pressing method reached 45.46%.That is to say,direct pressing method can be used as a simple and feasible method to extract 6-gingerol from ginger juice,and it also provides scientific basis and reference value for the feasibility of the industrialization of direct pressing ginger juice.The score of the simulation docking of PDE4 and 6-gingerol by MOE indicates that PDE4 is a potential target of anti-inflammatory effect of 6-gingerol.On this basis,PDE4 was selected as the target for further study on the anti-inflammatory effect of 6-gingerol;6-gingerol at 25-100 ?g/m L concentration had no effect on the viability of RAW264.7cells;LPS could activate RAW264.7 cells and significantly enhance the viability of RAW264.7 cells,while 6-gingerol at 25-100 ?g/m L concentration inhibited the viability of RAW264.7 cells in the activated state.The concentration of 6-gingerol in this experiment was determined to be 25-100 ?g/m L.Compared with the blank group,the secretion of TNF-? and IL-6 increased significantly and c AMP decreased significantly in LPS group,suggesting that 10 ?g/m L LPS could induce inflammation of RAW264.7cells,while 6-gingerol at 25-100 ?g/m L concentration inhibited the secretion of TNF-?and IL-6 in RAW264.7 cells induced by LPS,but promoted the secretion of c AMP,suggesting that 6-gingerol at this concentration could effectively inhibit inflammation of RAW264.7 cells and play a role.Anti-inflammatory effect;PDE4 content in inflamed RAW264.7 cells induced by LPS increased significantly,while PDE4 expression in inflamed cells treated with 6-gingerol at 25-100 ?g/m L concentration was significantly inhibited,suggesting that 6-gingerol has anti-inflammatory effect on RAW264.7 cells,and the anti-inflammatory mechanism may be that 6-gingerol inhibits the expression of PDE4 protein in inflamed cells.