| Rationales:Depression is a major disease that threatens human health.At present,there are many drugs for the treatment of depression,mainly imported chemical drugs,which have many problems such as adverse reactions,high price,and easy recurrence after drug withdrawal.Therefore,the market demand of new antidepressant drugs is huge.Chaigui granule,a new traditional Chinese medicine,is developed on the basis of Xiaoyaosan.Its suitable for mild and moderate depression and change the license,the depressive mood,depression,anxiety,tension and other core symptoms,memory and concentration,power loss,low desire,poor appetite,constipation,fatigue and somatic symptoms were improved.It is composed of 6 herbs of Bupleurum,Angelica,Peppermint,Licorice,Atractylodes,and White Peony,and has been clinically approved（2018L03149）.It is suitable for patients with mild and moderate depression and belongs to liver depression syndrome and has core symptoms such as depression,low mood,anxiety and tension,as well as inattention,loss of memory,decreased libido,poor appetite,Physical symptoms such as constipation and fatigue have a significant improvement effect.In our previous study,we have studied the in vitro chemical composition,in vivo and in vitro intestinal absorption characteristics,blood entry,brain entry and PK-PD combination of Chaigui Granules,and compared the blood entry and intestinal absorption characteristics of Chaigui Granules in normal and CUMS rats.The results showed that 45 components with significant absorption differences were found in the valgus intestinal sac experiment,and 10 components with significant absorption differences were found in the one-way intestinal perfusion experiment,suggesting that these components provided a material basis for the antidepressant effect,but the distribution,metabolism and excretion of its index components in normal and depressive states were not clear.Objective:In this study,the distribution,metabolism and excretion of Chaigui granules in and out of normal and CUMS rats were studied,and the process of Chaigui granules in vivo was clarified,so as to provide an important basis for metabolic screening of new drug research and efficacy evaluation of clinical drug use.Establish pharmacokinetic parameters curve of index components;The overall animal model was established to study the distribution of Chaigui granules in tissues;The metabolism of Chaigui granules in intestinal flora was studied in vitro.The overall animal model was established to study the excretion of Chaigui granules in urine and feces,so as to clarify the in vivo process of Chaigui granules,and provide important evidence for the metabolic screening of new drug research and the efficacy evaluation of clinical medication.Methods:1.UPLC-MS/MS technology was used to establish the pharmacokinetic curves of the index components of Chaigui granules,analyze their pharmacokinetic parameters,and compare their differences in pharmacokinetic behaviors under normal and CUMS conditions.2.The distribution of Chaigui granules in heart,liver,spleen,lung,kidney,ileum and hippocampus of normal and CUMS rats was determined by UPLC-MS/MS analysis.3.The intestinal flora incubation method was used to compare the metabolism of Chaigui Granules with that of normal and CUMS model rats under constant temperature anaerobic culture.4.The excretion of Chaigui granules in urine and feces of normal and CUMS rats was determined by UPLC-MS/MS and compared.Results:1.According to the previous study of our research group,61 prototype components were found in the serum of normal and CUMS rats,of which 12prototype components were identified in the common components of normal and CUMS groups,namely gallic acid,chlorogenic acid,vanillic acid,ferulic acid,paeoniflorin,paeoniflorin,liquiritin,liquiritin,Z-butenylphthalide,n-butylphthalide,glycyrrhetinic acid,and isoliquiritin.Through the analysis of pharmacokinetic curves and parameters in normal and depressive state,the results showed that the curves of ferulic acid,chlorogenic acid,paeoniflorin,liquiritigenin,Z-butenylphthalide and n-butylphthalide in the model group and the blank group were different.The curves were three peaks,and the peak time was 0.17 h,4 h and 12 h,respectively.The pharmacokinetic curves of liquiritin,glycyrrhetinic acid,isoliquiritin,vanillic acid,gallic acid and paeoniflorin in normal and model groups were similar.The retention time and elimination time of chlorogenic acid,ferulic acid and isoliquiritin in CUMS rats increased under depression and the absorption of paeoniflorin increased in vivo.2.Nine prototype components were identified by UPLC-MS in different tissues of normal and CUMS rats.Chlorogenic acid,paeoniflorin,liquiritin,liquiritin,isoliquiritin and 5-hydroxylliquiritin were widely distributed in normal and CUMS rats,while saikosaponin a was only detected in the ileum of normal and CUMS rats,which may affect its distribution in rat tissues or fail to be detected in other tissues due to its low concentration.Atractylenolide I,saikosaponin c and 5-hydroxylliquiritin were detected in hippocampus,indicating that they could enter into hippocampus through blood-brain barrier.The distribution of these components in different tissues of normal and CUMS rats was different,indicating that the binding force between drugs and tissues in vivo was changed in depression.3.Chaigui granules were incubated with normal rat intestinal bacterial fluid and CUMS model rat intestinal bacterial fluid under constant temperature anaerobic.Compared with blank samples,32 prototype compounds were identified in normal and CUMS model rat intestinal bacterial fluid,of which 31 were common.E-ligustilide was only detected in normal rat intestinal bacterial fluid.The results showed that most of the chemical components of Chaigui Granules were reduced by intestinal flora metabolism after incubation with intestinal flora in rats.The representative components were saikosaponin a,c,d,glycyrrhizic acid,liquiritin,paeoniflorin,paeoniflorin,atractylenolide III,butenylphthalide and ferulic acid.However,the contents of glycyrrhetinic acid and isoliquiritigenin in the intestinal flora of normal and CUMS rats increased with time.It was speculated that part of glycyrrhetinic acid might be formed by the metabolism of glycyrrhizic acid in the intestinal flora of rats,but its content in the intestinal flora of normal rats was significantly higher than that of CUMS rats.The metabolic rates of glycyrrhizic acid,paeoniflorin and saikosaponin a in the intestinal flora of normal rats were significantly higher than those of CUMS rats.4.A total of 23 prototype compounds were identified by LC-MS in the urine of normal and cums model rats,and 40 prototype compounds were identified in the feces of normal and cums model rats.The cumulative excretion rates of these compounds in urine and feces of normal and cums rats over time were studied.The excretion of triterpenoid saponins in the urine of blank rats was greater than that of cums rats,and the excretion of flavonoids and lactones in the urine of cums rats was greater than that of blank rats;the excretion of organic acids,terpenoids and flavonoids in the feces of normal rats was greater than that of cums rats,and the excretion of triterpenoid saponins,saikosaponins and lactones in the feces of cums rats was greater than that of normal rats.Conclusion:1.Through the comparative study of pharmacokinetic curves and parameters of 13 chemical components in Chaigui Granules in normal and CUMS rats,AUC（0–∞）and MRT（0–∞）of all components in CUMS rats increased,indicating that the absorption and retention time of these components in cums rats are increased under depression,so as to prolong the effective time and enhance the effect.2.Using UPLC-MS technology,nine prototype components were identified in different tissues of normal and CUMS rats after single intragastric administration of Chaigui granules,and the distribution differences of nine chemical components in Chaigui granules in different tissues of normal and CUMS rats were compared.It not only shows that these nine components may be antidepressant active components,which further confirms that the effective parts or target organs of Chaigui granules are hippocampus and liver tissue.3.A total of 32 compounds were identified by culturing Chaigui granules in normal and CUMS rat intestinal microflora.The changes of these compounds in normal and CUMS rat intestinal microflora at different incubation times were compared.The results showed that most of the chemical components of Chaigui granules were reduced by intestinal microflora metabolism after incubation with rat intestinal microflora.Only the contents of glycyrrhetinic acid and isoliquiritigenin in normal and CUMS rat intestinal microflora increased with time.4.A total of 23 compounds were identified by LC-MS in the urine of normal and cums model rats.Compared with the blank group,12 compounds were significantly different;a total of 42 compounds were identified in feces,of which 22were significantly different.The cumulative excretion rates in feces of normal and CUMS model rats were compared.The results showed that under depression,the excretion amount and excretion time of chemical components in Chaigui granules in urine and feces were affected,and the excretion process in vivo under physiological and pathological conditions was revealed. |
PDF Full Text Request