To Investigate The Effects Of Acupuncture Combined With HRT On Sex Hormone-related Metabolites And Metabolic Pathways In POI Patients Based On Metabolomics

Objective:To explore the optimal acupuncture acupoint compatibility scheme for patients with POI,based on the data mining technology of ancient and modern literature.To further observe the clinical efficacy and safety of acupuncture combined with Hormone replacement treatment(HRT)through randomized controlled clinical trials.To explore the mechanism of acupuncture and HRT regulating serum sex hormones in patients with POI from the perspective of metabonomics by using metabonomics technology.Methods:The database of acupuncture treatment prescription in ancient and modern literature about Premature ovarian insufficiency(POI)was established respectively,the rules of meridian selection and acupoint selection were analyzed by EXCEL and SPSS Modeler 19.0 software,and a optimized acupuncture prescripition was obtained.In a further randomized controlled clinical trial,64 patients with premature ovarian insufficiency were randomly divided into three groups: patients in acupuncture plus HRT group treated by acupuncture plus HRT,control group 1 treated by acupuncture and control group 2 treated by HRT.The course of treatment in the three groups was 3 months.The serum anti-Mullerian hormone(AMH),serum sex hormones(FSH,LH,and estrogen),the number of antral follicles,modified Kupperman score,MENqol score and SAS score were evaluated before and after treatment for therapeutic effect evaluation by three curative methods and the adverse reactions was recorded,and blood routine,liver function and renal function were detected before and after treatment for safety evaluation.Finally,12 patients with POI in acupuncture group,9 patients in HRT group and 10 patients in acupuncture combined with HRT group were randomly selected for detection of metabolites.The serum samples of POI patients in three groups before and after treatment were collected.LC-MS technique was used to detect serum differential metabolites and Metabo Analyst 4.0was used to analyze the metabolic pathway.Results:1.A total of 102 acupuncture prescriptions were collected in ancient literature,covering 40 acupoints,while 106 acupuncture prescriptions were collected in modern literature,covering 75 acupoints.Meridian Selection: in both ancient and modern literature,Ren meridian is the main selection of Meridian,followed by Kidney Meridian,Bladder Meridian and Spleen Meridian,more selection of Stomach Meridian in modern literature.Part Selection: in both ancient and modern literature,abdominal area is the most important part,followed by foot area,leg area,waist area and sacral area,more selection of dorsal region in modern literature.The choice of acupoints: Guanyuan is the first high-frequency acupoint in both ancient and modern literature,the other high-frequency acupoints are quite different,Zhongji,Yongquan,Yinjiao and Shangqiu in ancient data,while Sanyinjiao,Shenyu,Zigong and Zusanli in modern data;The compatibility of ancient and modern high-frequency acupoints is mainly based on Guan Yuan,and the other compatibility acupoints are quite different.Selection of specific points: Ancient and modern literature high-frequency specific points are Mu points,five Shu points,crossing point,back Shu points.The ancient literature selects the eight confluence point also,while the modern literature selects the Yuan-Source point.The frequency of specific acupoints in both ancient and modern literature is Guanyuan,followed by Zhongji,Yinjiao,Yongquan,Shangqiu and Ran Gu in ancient literature,and Guanyuan,Sanyinjiao,Shenyu,Zusanli,Taixi,and Taichong in modern literature.The compatibility of specific acupoints is mainly based on Guanyuan in both ancient and modern times,while the other compatibility of specific acupoints is quite different.2.The levels of AMH in acupuncture group after treatment was higher than that before treatment.the level of both FSH and LH decreased,level of E2 increased,and socres of modified KI,MENqol,and SAS declined after treatment in three groups,respectively.The improvement of both estradiol and AMH in acupuncture group were better than those in hormone group,respectively.The effect of acupuncture on improving estradiol was more stable when the level of estrogen was higher than 22pmol/L or the level of FSH was lower than 40U/L before treatment.The effect of acupuncture plus HRT on the sexual life was better than that of acupuncture and HRT group,respectively.3.There were 5 metabolites in Acupuncture group before and after treatment: l-glutamic acid,D-proline,?-aminopropionitrile,citric acid,N-acetyl-L-phenylalanine.There are 8 main metabolic pathways: D-glutamate and D-glutamate metabolism,arginine and proline metabolism,arginine biosynthesis,aminoacyl-t RNA synthesis,cyano amino acid metabolism,tricarboxylic acid cycle,glyoxylic acid and dicarboxylic acid metabolism,phenylalanine metabolism.In HRT group,there were 6 different metabolites before and after treatment: l-lysine,?-alanine,5-aminopentanal,?-linolenic acid,deoxycholic acid and creatinine,and there were 10 metabolic pathways: lysine degradation,lysine production,biotin metabolism,pantothenic acid and coenzyme A biosynthesis,?-alanine metabolism,proline metabolism,linolenic acid metabolism,pentose phosphate pathway,arginine and proline metabolism.There were two different metabolites before and after treatment: lmethionine and 4-aminobutyraldehyde dimethyl acetal,and four metabolic pathways were involved: ?-alanine metabolism,arginine and proline metabolism,cysteine and methionine metabolism,and aminoacyl-t RNA synthesis.Conclusion:1.The prescriptions' selection of acupuncture and moxibustion treatment in ancient and modern literature on POI are roughly the same,while there are some differences between them,the modern developed on the basis of inheritance.2.Three therapies can improve the levels of serum sex hormones,symptoms,quality of life and anxiety of POI patients.Acupuncture can improve the ovarian reserve and levels of estrogen.Especially for patients with a certain level of estrogen or FSH levels less than40U/L,acupuncture has a better effect on improvement of estrogen.Acupuncture combined with HRT has a synergistic effect on improving sexual life.The short-term adverse reactions of the three therapies were low.3.Acupuncture combined with hormones,acupuncture and HRT regulate metabolites and metabolic pathways differently.