Study On Acorus Tatarinowii Schott In Dingzhi Pills On "Brain Targeting" Of Five Saponins

Objective:Dingzhi pill is composed of ginseng,poria cocos,radix polygalae and Acorus gramineus,according to the quality ratio of 3:3:2:2,initially recorded in the Chinese ancient book Bei-Ji-Qian-Jin-Yao-Fang compiled by Sun Si-Mao.It is a basic prescription for the treatment of alzheimer's disease in traditional Chinese medicine,and is still used in clinical practice.Ginsenosides of the main medicinal material in ginseng acting as“monarch herb”in Dingzhi pills,tenuifolia of the main medicinal material in ginseng acting as“ministerial herb”in Dingzhi pills,their main active ingredients are saponins,but these saponins have low oral bioavailability and are difficult to be absorbed by gastrointestinal tract.Acorus tatarinowii Schott,as“adjuvant drug”in Dingzhi pills,has the effects of resolving dampness,having appetite and inducing medicine upward.The research group confirmed that Acorus tatarinowii Schott can promote the intestinal absorption of ginsenosides in Dingzhi pills.Since alzheimer's disease is a brain disease,whether Acorus tatarinowii Schott can promote the"brain-targeted"properties of ginsenosides to reach the target organs is worth studying.In this paper,the effect of Acorus tatarinowii Schott on the five saponins in Dingzhi pills was explored from the perspective of promoting"brain targeting",which can provide scientific basis for the clinical medication of this prescription and reference for the research on compound compatibility.Methods:1.The mouse pharmacokinetic model was established by intragastric administration of different prescriptions of Dingzhi pills,and the plasma was detected by ultra performance liquid chromatography-electrospray ionization-mass spectrometry?UHPLC-ESI-MS/MS?.And the concentration of ginsenoside Rg1,Re,Rb1,Rd and tenuifolin in brain homogenate,DAS2.0 software fitting the pharmacokinetic parameters of mice,comparing the five saponins of different groups pharmacokinetic parameters such as AUC_0-t,C_max,t_1/2.According to the formula,the concentration ratio?Ce?of volatile oil of Acorus tatarinowii Schott,the relative uptake rate of brain?Re?and the targeted targeted index?DTI?were calculated to evaluate the targeting of five saponins.2.After oral administration of different concentrations of volatile oil and verapamil,the content of rhodamine 123 in plasma and brain tissue of mice was detected by fluorescence spectrophotometry,and absorbed by P-gp substrate rhodamine 123 in mice.The situation,indirectly explore the effect of volatile oil of Acorus tatarinowii Schott on P-gp efflux.3.After the mice were intragastrically administered with different concentrations of volatile oil and verapamil,the expression of P-gp in jejunum and brain tissue of mice was detected by Western blot method,and the mechanism of volatile oil absorption by Acorus tatarinowii Schott was preliminarily clarified.The molecular docking software was used to score 41 constituents of gladiolus tatarinowii volatile oil with P-gp,and the effect of constituents in gladiolus tatarinowii volatile oil on p-gp was evaluated.Results:1.The contents of ginsenosides Rg₁,Re,Rb₁,Rd and tenuifolin AUC_0-t in the whole prescription group of Dinzhi pills in plasma were increased by 165.5%,237.5%,225.9%,248.8%and 238.4%?P<0.05?,respectively,and the contents of C_maxax were increased by 160.9%,139.5%,216.7%,250.4%and 223.6%?P<0.05?.T_1/2 was 3.09,2.57,1.67,1.76 and 2.43 times as much as that of the Acorus tatarinowii Schott rhizome group?P<0.05?,and MRT_0-?was 1.35,1.19,2.13,2.56 and 2.44 times as much as that of the Acorus tatarinowii Schott rhizome group?P<0.05?.In brain tissue,ginsenosides Rg1,Re,Rb1,Rd and tenuifolin AUC0-t in the whole prescription group of Dingzhi pill were 5.57,3.84,3.30,4.20 and 4.10 times of that in the loss of Acorus tatarinowii Schott prescription group,respectively?P<0.05?.Cmax was 2.25,1.45,2.16,2.28 and2.30 times of the loss of Acorus tatarinowii Schott group?P<0.05?.T_1/2 and MRT_0-?infinity were also significantly increased compared with that in the loss of Acorus tatarinowii Schott group?P<0.05?.The above results indicated that Acorus tatarinowii Schott could promote the absorption of five saponins in vivo and the absorption was concentration dependent.At the same time,DTI of ginsenosides Rg1,Re,Rb1,Rd and tenuifolin were 2.10,1.14,1.01,1.20 and 1.21,respectively.The results showed that Acorus tatarinowii Schott could promote the absorption of five saponins in vivo in a concentration-dependent manner,indicating that acorus tatarinowii had brain targeting effect on the absorption of five saponins.2.Mouse were given different doses of Acorus tatarinowii Schott volatile oil and verapamil.After 1 h,the cumulative amount of rhodamine 123 in plasma and brain tissue of the medium and high dose volatile oil groups was significantly higher than that of the blank control group?P<0.05?,and the K_p value was 1.36,1.48 and 1.21 times as high as that of the blank control group?P<0.05?.After 2 h of intragastric administration,K_p values in the medium-dose and high-dose volatile oil groups were 1.79,2.24 and1.66 times as high as those in the control group?P<0.05?.The above results showed that the volatile oil of Acorus tatarinowii Schott volatile oil could increase the accumulation of rhodamine 123 in mice in a dose-dependent manner,suggesting that the volatile oil of Acorus tatarinowii Schott could promote drug absorption by inhibiting P-gp efflux.3.Compared with the blank control group,the volatile oil group of Acorus tatarinowii Schott can significantly reduce the expression level of P-gp in the intestinal tract and hippocampal area of brain tissues of mice?P<0.05?,indicating that the volatile oil of Acorus tatarinowii Schott can inhibit the expression of P-gp.According to the molecular docking scoring,the combination of?-asarone,?-asarone and?Z,Z?-9,12-octadecanedienoic acid was better,and the combination scoring was-15.2863,-13.1868 and-13.0812,respectively,which were mainly combined with P-gp through conjugated double bonds and hydrogen bonds.Conclusion:1.The volatile oil of Acorus tatarinowii Schott can promote the absorption of ginsenosides Rg1,Re,Rb1,Rd and tenuifolin in Dingzhi pills in mice in a concentration dependent manner.At the same time,Acorus tatarinowii Schott has brain targeting effect on five saponins.2.The volatile oil of Acorus tatarinowii Schott can increase the accumulation of rhodamine 123,the P-gp substrate in mice,suggesting that the promoting effect of Acorus tatarinowii Schott may be related to the inhibition of P-gp3.The volatile oil of Acorus tatarinowii Schott can inhibit the expression of P-gp in the jejunum and hippocampus of mice,suggesting that inhibiting the expression of P-gp may be one of the main reasons that Acorus tatarinowii Schott promotes the absorption of saponins in vivo.