| Hypertension is one of the most common chronic diseases in the world,and it is the main cause of cardiovascular disease and premature death.In recent years,the prevalence of hypertension in my country has increased.Food and medicine homologous substances have mild medicinal properties and strong medicinal effects.They are often used in the treatment of chronic diseases and play an important role in the treatment of diseases such as hypertension,hyperlipidemia,and hyperglycemia.This article screened and studied the antihypertensive effects of the food and medicine homologous substances,focusing on the antihypertensive effects of Eucommia ulmoides leaves,Fructus Crataegi,and Sophora japonicus,and optimized the extraction process of chlorogenic acid of E.ulmoides,and carried out related researches on the antihypertensive mechanism of chlorogenic acid.It provides scientific basis and theoretical support for the development and application of E.ulmoides leaf.The main content of this research includes the following 4 aspects:1.Research on the anti-hypertensive effect as well as mechanism of medicinal and edible plant.The medicinal and edible plants,which possess the anti-hypertensive effect,are screened out through network pharmacology analysis.Animal experiment are carried out to verify the significant anti-hypertensive effect of a variety of medicinal and edible plants.Meanwhile,the main components for anti-hypertensive in medicinal and edible plants are subsequently analyzed.The results showed that 40 plants with excellent anti-hypertensive effects are selected out form 108 medicinal and edible plants.Among them,Eucommia ulmoides leaves,Fructus Crataegi and Sophora japonicus revealed significant antihypertensive effects.Animal experiments certified that these 3 plants could significant reduce the level of systolic blood pressure,Ang II and ALD in SHR rats and increase plasma NO,especially for Eucommia ulmoides leaf.2.Study on the extraction process of chlorogenic acid from E.ulmoides leaf with natural deep eutectic solvent.In order to screen the natural deep eutectic solvent system with excellent extraction efficiency,many factors are investigated,including ultrasonic power,ultrasonic time,ultrasonic temperature,water content in natural deep eutectic solvent,and molar the ratio of each component in natural deep eutectic solvent.Their effects on chlorogenic acid extraction are also analyzed.On the basis of single-factor experiments,3 factors(water content in natural deep eutectic solvent,ultrasonic time,and ultrasonic power)are found to significantly affect the extraction yield,and these 3 factors are selected for response surface optimization.The antioxidant activities of the crude chlorogenic acid extract of E.ulmoides leaf are further investigated.The results show that the factors affecting the yield of chlorogenic acid range as water content in natural deep eutectic solvent > ultrasonic time > ultrasonic power > moles of natural deep eutectic solvent components >ultrasonic temperature.According to the results of response surface optimization and the actual need for production,the optimal extraction process conditions are designated as follows: ultrasonic time,42 min,ultrasonic power,340 W;water content in eutectic solvent,5 mol.Under optimal extraction conditions,the extraction yield of chlorogenic acid reach 31.46 mg/g,which is 18.18% and 14.81% higher than the extraction yield of water extraction(26.61 mg/g)and ethanol extraction(27.40mg/g),respectively.The chlorogenic acid crud extract reveal good antioxidant activities,and its free radical scavenging effect is positively correlated concentration within a certain range.In order to further separate and purify chlorogenic acid,the medium pressure liquid chromatography is used for extraction based on the above selected optimal extraction conditions after crub extract purified by resin.The preparation process is as follows: column temperature,25℃;detection wavelength,254 nm;flow rate,15 m L/min;mobile phase,sample size,1 m L,1% phosphoric acid and alcohol(v:v,90:10).The obtained product is analyzed by high performance liquid chromatography,and the purity of chlorogenic acid is 95.67%.3.The antihypertensive mechanism network pharmacology analysis of chlorogenic acid from E.ulmoides leaf.6 main components with anti-hypertensive effects were screened out from Eucommia ulmoides leaves by network pharmacology analysis and 116 targets of the main components were also located.Among these targets,58 of them were of the targets of chlorogenic acid.Among them,21 targets are hypertension-related targets.Besides,HPLC analysis had verified that the content of chlorogenic acid in Eucommia ulmoides leaves was kept at a relative high level.The screened targets are further analyzed by the DAVID database and map into 67 GO terms.Ten KEGG pathways are significantly enriched and VEGF signal pathway,PI3K-Akt signaling pathway and RELAXIN signaling pathway are screened out,which are relate to the antihypertensive of chlorogenic acid and involvement with the biochemical process of protein kinase phosphorylation,vascular endothelial dysfunction,cell metabolism and oxidative stress.Based on the PPI network analysis,6 key targets are identified,including CASP3,KDR,ACE,MMP2,ERBB2 and ABCB1.Molecular docking analysis confirm that chlorogenic acid can bind tightly to these key targets.The results indicate that chlorogenic acid can take function in antihypertensive through multiple targets and multiple pathways.4.Animal experiments to verify the antihypertensive mechanism of chlorogenic acid from E.ulmoides leaves.Spontaneously hypertensive rats(SHR)are used as model and chlorogenic acid of E.ulmoides leaves is fed uninterruptedly.After 5 w feeding,the height ratio of heart,liver,and kidney are measured.The NO,Ang II,ALD,ET-1,IL-6,and TNF-α in plasma are determined by ELISA.The results show that chlorogenic acid cause systolic and diastolic blood pressure in the tail artery of vascular gradually decrease in SHR rats.The ratio of heart and liver also decrease.Meanwhile,the vascular-related factors ANGII,ALD,and ET-1 will significant decrease and NO level will significant increase.At the same time,the pro-inflammatory factors IL-6 and TNF-α are also significantly reduced.It can be concluded that the antihypertensive effect of chlorogenic acid may mainly affect the key targets,e.g.KDR,ACE,ERBB2 and ABCB1,as well as key pathways,e.g.VEGF pathway and P13K-Akt pathway.Therefore,the antihypertensive effect of chlorogenic acid can be achieved from multiple targets and multiple pathways.In addition,chlorogenic acid can also help improve the antihypertensive effect through anti-inflammatory effects and the protecting heart/liver effects.The above mentioned results can deeply explain the antihypertensive mechanism of chlorogenic acid from E.ulmoides leaf,and provide more scientific basis for the high value conversion and the industrial application of chlorogenic acid from E.ulmoides leaf as antihypertensive products. |
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