| Aging refers to the general weakening of various functions,degenerative changes of tissues and organs,progressive decrease of physiological adaptability to the environment and comprehensive performance of a variety of physiological or pathological processes.Traditional Chinese medicine generally believes that kidney is closely related to aging,and kidney tonifying herbs can delay aging in theory,but the anti-aging mechanism of traditional Chinese medicine is still unclear.Liuwei Dihuang pill and Jingui Shenqi Pill have been used for kidney tonifying for a long time in clinical practice,with significant curative effect and widely recognized.Liuwei Dihuang Pill originated from the Song Dynasty Taiyi Qian Yi's "Xiao Er Yao Zheng Zhi Jue",which is composed of 6 traditional Chinese medicine,namely,Radix Rehmanniae Preparata,Fructus Corni,Cortex Moutan,Rhizoma Dioscoreae,Poria cocos and Rhizoma alismatis.It has the effect of "nourishing kidney yin and filling kidney essence".Jingui Shenqi Pill is from the “Jin Kui Yao Lue”,written by Zhang Zhongjing,a medical saint.It is made up of 8 medicinal materials including Aconitum carmichaelii,Cinnamon Twig,Rehmannia glutinosa,Fructus Corni,yam,Cortex Moutan,Poria and Rhizoma alismus.The prescription is highly precise and highly praised by all generations of doctors,and is honored as the ancestor of tonifying kidney and antiaging agents.Therefore,in-depth study on the anti-aging effect of Liuwei Dihuang pill and Jingui Shenqi Pill can not only clarify the mechanism of anti-aging effect of Liuwei Dihuang pill and Jingui Shenqi Pill,but also provide scientific basis for their clinical application,so as to expand their application in anti-aging and make the society happy.In order to further understand the material basis and mechanism of anti-aging effect of Liuwei Dihuang pill and Jingui Shenqi Pill,the chemical composition in vitro and in vivo of Liuwei Dihuang pill and Jingui Shenqi Pill were analyzed,and the differences of active components between them were discussed;The anti-aging mechanism of the two prescriptions was predicted;Through the establishment of acute aging model induced by D-galactose,the anti-aging pharmacological effect and mechanism of the two prescriptions were verified;LC-MS combined with cluster analysis,CA,PCA and OPLS-DA were used to study the difference of potential effective components in Liuwei Dihuang concentrated pills and water-honeyed pills,and the potential effective components with anti-aging effect in different formulations of Liuwei Dihuang pills were gradually screened and speculated.The main contents of this paper are as follows:1.Study on the pharmacodynamic material basis of anti-aging effect of Liuwei Dihuang pill and Jingui Shenqi PillThe chemical constituents of Liuwei Dihuang pills and Jingui Shenqi pills were analyzed and identified by UPLC-QE-Orbitrap-MS.A total of 99 compounds(28triterpenoids,18 iridoid glycosides,10 monoterpenoids,and other sesquiterpenoids,phenylpropanoids,flavonoids,etc.)were identified in Liuwei Dihuang pills,among which 65 compounds have been reported,The other 34 compounds were identified from Liuwei Dihuang pills for the first time.A total of 112 compounds(8 alkaloids,23 triterpenoids,16 iridoid glycosides,9monoterpenoids,and other sesquiterpenoids,amino acids,phenols,phenylpropanoids,flavonoids,etc.)were identified in Jingui Shenqi pills.Among them,100 compounds have been reported,and the remaining 12 compounds were identified from Jingui Shenqi pills for the first time.Liuwei Dihuang pill is the reduced prescription of Jinkui Shenqi Pill.According to the requirements of indications,Fuzi and Guizhi are reduced in the prescription.At the same time,88 compounds in Jingui Shenqi Pill are common with Liuwei Dihuang pill,11 compounds are characteristic components of Aconitum carmichaelii in Jingui Shenqi Pill,13 compounds are characteristic components of Ramulus Cinnamomi in Jingui Shenqi Pill,and 3 compounds(quercetin,protocatechuic acid,protocurcumenol)are common compounds of Cornus officinalis and Ramulus Cinnamomi.The results can provide the basis for the difference analysis of serum chemical components and network pharmacology research of Liuwei Dihuang pills and Jingui Shenqi pills.2.Study on serum pharmacochemistry of Liuwei Dihuang pill and Jingui Shenqi Pill in ratsUPLC-Q-To F-MS was used to detect the chemical components in the serum of rats treated with Liuwei Dihuang pill and Jingui Shenqi Pill.By comparing the chemical components in serum and in vitro of rats after administration of Liuwei Dihuang pill,32 components were identified successfully.By comparing the chemical components in serum and in vitro of rats after administration of Jinkui Shenqi pill,31 components were identified successfully.Furthermore,by analyzing the differences of blood components between the two prescriptions,18 compounds were the unique blood components of Liuwei Dihuang pills,17 compounds were the unique blood components of Jingui Shenqi pills,and 14 compounds were the common blood components of the two prescriptions.From the point of view of serum pharmacochemistry,the above research contents studied the pharmacodynamic material basis of Liuwei Dihuang pill and Jingui Shenqi Pill.The research results can provide the basis for the network pharmacology research of Liuwei Dihuang pill and Jingui Shenqi Pill.3.Network pharmacology was used to predict the anti-aging mechanism and potential effective components of Liuwei Dihuang pill and Jingui ShenqiThe related information of active components,potential targets and pathways in Liuwei Dihuang pill and Jingui Shenqi Pill were obtained by means of network pharmacology:(1)in Liuwei Dihuang pill and Jingui Shenqi Pill,51 and 62 compounds have good pharmacokinetic activities,respectively,which can be used as candidate compounds for anti-aging;(2)4612 and 5140 target proteins can be used as potential therapeutic targets for Liuwei Dihuang pill and Jingui Shenqi Pill,respectively;(3)Liuwei Dihuang pill is mainly involved in the biological process of G protein coupled amine receptor and vasoconstriction;Jingui Shenqi Pill is mainly involved in G protein coupled neurotransmitter receptor,acid secretion,negative regulation of cell secretion,activity of somatostatin receptor,positive regulation of vasoconstriction and positive regulation of smooth muscle contraction;(4)The potential pharmacodynamic components in Liuwei Dihuang pills were identified from three aspects: the source pathway of chemical components,pharmacokinetics and pharmacodynamic correlation.Finally,it was speculated that seven chemical components were potential pharmacodynamic components of Liuwei Dihuang pills.From the perspective of the source of the compound,the potential effective components of Liuwei Dihuang pill and Jingui Shenqi Pill can represent the integrity of the compound.4.Verification of anti-aging mechanism of Liuwei Dihuang pill and Jingui Shenqi PillThe acute aging mouse model was established by D-galactose.The key indicators' levels of GSH-Px and NF-?B-p65 in serum and kidney/brain tissue were detected by WB,PCR and IHC and potential target proteins were determined.The results showed that: the SOD,MDA and GSH-Px values of the treatment group were significantly different from those of the model group(P< 0.01);Telomere length in high dose group was significantly increased(P< 0.01);The levels of TNF-? in serum,brain tissue and kidney tissue of middle and high dose groups were significantly decreased(P< 0.05);The content of IL-1 in serum,kidney and brain of middle and high dose group was significantly decreased(P< 0.05);The expression of NF-?B-p65 in middle and high dose groups was significantly decreased(P< 0.01),and it was dissociated from cytoplasm to nucleus.It is suggested that Liuwei Dihuang pill and Jinguishenqi pill can improve the scavenging ability of oxygen free radicals,improve the anti-oxidation ability and slow down the cell telomere shortening by participating in the biological process of G protein coupling receptor activity and improving the vasoconstriction,thus playing an anti-aging pharmacological role.Based on the above research results,the scope of this study can be extended to predict the potential effective components of Liuwei Dihuang pills with different formulations,and explore the influence of different preparation processes on the potential effective components.5.Speculation on the potential effective components of different dosage forms of Liuwei Dihuang pillsThe contents of 10 chemical components in Liuwei Dihuang concentrated pills and Shuimi pills were determined by UPLC-Qq Q-MS.Furthermore,by using CA,PCA and OPLS-DA multivariate statistical analysis methods,combined with the results of in vitro and in vivo component detection and network pharmacology of Liuwei Dihuang pills,the content differences between concentrated pills and Shuimi pills were comprehensively evaluated and compared.It is speculated that paeoniflorin,Rehmannioside D and benzoylpaeoniflorin are the potential effective components in Liuwei Dihuang pills of different dosage forms.In this paper,the chemical constituents of Liuwei Dihuang pill and Jingui Shenqi Pill in vitro and in vivo were systematically studied,which laid a foundation for the screening of their effective substances.Through the network pharmacology and main pharmacodynamic experiments,the anti-aging mechanism and effective substance basis of Liuwei Dihuang pill were preliminarily speculated,which provided a theoretical basis for their scientific and rational drug use in the future. |
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