| Objective To explore the connotation of "Spleen-Ying-Mai correlation" theory and its internal correlation with Atherosclerosis(AS),and seek corresponding treatment methods and prescriptions to provide theoretical basis for the treatment of AS.To explore the drug safety of Jian-Pi-Tong-Mai formula(JPTMF),so as to provide experimental basis for clinical rational drug use,and explore the potential pharmacodynamic substances of JPTMF,so as to provide preliminary basis for improving the safety research on the basis of pharmacodynamic substances of this formula.Methods 1.Theoretical research:By referring to ancient and modern literature,clarify the connotation of "Spleen-Ying-Mai correlation" theory and its internal correlation with the incidence of AS,and the corresponding treatment principles and representative prescriptions are put forward.The scientificity of JPTMF was demonstrated through prescription analysis and modern pharmacodynamics and therapeutic mechanism study.2.Experimental research:2.1 Safety evaluation of different doses of JPTMF in two animals.(1)Preliminary safety experiment of JPTMF effective dose for 21 days.C57BL/6J mice was selected as experimental subjects.Twenty mice were randomly divided into blank control group and JPTMF group with ten mice in each group by random number table method.Mice in JPTMF group were given the effective dose of JPTMF for 21 days by gavage,.and the mice in blank control group were given the same amount of distilled water.The safety of effective dose of JPTMF was preliminatively evaluated by comparing the effects of the drug on the general condition,body weight and main organs of mice with those of blank control group.(2)JPTMF acute toxicity experiment.Kunming mouse(KM mouse)was selected as experimental subjects.First,the maximum dose of JPTMF was determined by pretest,and then the maximum dose experiment was carried out.According to random number table method,twenty four mice were randomly divided into blank control group and JPTMF group,with twelve mice in each group and half male and half female.The maximum dosage of JPTMF group was given intragaadministratively within 24 h,and the general condition,food intake,water intake and body weight of KM mice were observed.After 14 days of observation,automatic blood biochemical analyzer was used to detect the blood biochemical indexes of KM mice and calculate the organ index of mice,so as to explore the toxic and side effects of the maximum dosage of JPTMF.2.2 Study on potential pharmacodynamic substances of JPTMF.The in vitro chemical constituents of JPTMF and its metabolites into blood and tissues were detected by ultraperformance liquid chromatography coupled with quadrupole time of flight mass spectrometry(UPLC-Q-TOF/MS)technology combined with UNIFI software,and the potential pharmacodynamic substances of JPTMF were preliminarily identified.Results 1.Theoretical research results:the core connotation of "Spleen-Ying-Mai correlation"theory is that the spleen is the foundation of the "Ying" and "Mai ",full spleen qi is the source of blood,only when the blood is stable and smooth can the pulse channels be smooth.The occurrence and development of AS is related to the disorder of "Spleen-Ying-Mai".Spleen qi deficiency is the beginning of the disease,"Ying-Mai " disharmony is the core of the disease,qi deficiency and blood stasis throughout AS disease.Invigorating spleen qi,promoting blood circulation and removing blood stasis are important methods to prevent and treat AS.JPTMF is a representative prescription for treating AS from the spleen based on the theory of "SpleenYing-Mai",and its prevention and treatment of AS is reasonable and scientific.2.Experimental research results:2.1 The effective dose of JPTMF 21 d preliminary safety experiment results showed:(1)During 21 day of oral administration of the effective dose of PTMF,the general condition of the mice in the JPTMF group did not show any abnormalities.(2)No significant difference in the weekly body weight of mice in the JPTMF group compared to the blank control group(p>0.05).(3)No significant difference in the organ indices of the main organs of the JPTMF group compared to the blank control group(p>0.05).(4)No significant pathological changes were observed in the heart and other major organs in JPTMF group.2.2 JPTMF acute toxicity test results showed:(1)Compared with blank control group,JPTMF group showed no abnormal general conditions during the maximum dosage of JPTMF administration within 24 h and the observation period within 14 d.(2)Compared with blank control group,JPTMF group had no significant difference in food intake and water intake(p>0.05),and no significant difference in body weight gain(p>0.05).(3)Compared with blank control group,the levels of liver function indicators ALT,ALB,TBIL and TP,blood lipid indicators CHOL,kidney function indicators BUN and CREA,and skeletal muscle and cardiac function indicators CK in serum were not significantly changed in the JPTMF group(p>0.05),the levels of liver function indicator ALP and lipid indicator TG in serum were significantly lower in the JPTMF group(p<0.05).(4)Compared with blank control group,there were no significant differences in organ indexes of heart,liver,spleen,lung,kidney,pancreas,testis,uterus and other organs in JPTMF group(p>0.05).2.3 JPTMF in vitro component analysis results showed:68 chemical components were detected in JPTMF granules.The compounds in the results,such as Salvianic acid A,Codonopsine,Caffeic acid,Calycosin,Naringenin,Salvianolic acid B,Chlorogenic acids,Glycyrrhizic acid all showed good anti-AS effect,and may be the pharmacodynamic substances of JPTMF against AS.2.4 Analysis of metabolites of JPTMF into blood and tissues showed that:78 metabolites were detected.Among them,46 metabolites entered the spleen,27 metabolites entered the plasma and 12 metabolites entered the blood vessels.Conclusion 1.Based on the theory of "Spleen-Ying-Mai correlation",it is clarified that spleen qi deficiency and disharmony of "Ying-Mai" may be the core pathogenesis of AS,and invigorating spleen and qi and promoting blood circulation and removing blood stasis is one of the important treatment methods of AS.JPTMF is a representative prescription of treating AS from spleen under the guidance of "Spleen-Ying-Mai correlation" theory,and its treatment of AS is reasonable and scientific.2.Safety studies of JPTMF at different doses in two kinds of animals showed that JPTMF was safe.Studies on potential pharmacodynamic substances of JTPMF preliminarily confirmed that Calycosin,Naringenin,Salvianolic acid B,Chlorogenic acids,Glycyrrhizic acid prototype and metabolites are potential pharmacological substances of this formula,and have good safety. |
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