The Discussion Of Lingnan Luo’s Gynecology On Drug Rules And The Network Pharmacology Of Yijing Decoction In The Treatment Of POI

Background and objects:To systematically evaluate the efficacy and safety of tonifying kidney therapy for premature ovarian insufficiency（POI）through Meta analysis.Data mining technology was used to study the medication law of Lingnan Luo’s gynecology in the treatment of POI to provide reference for the use of traditional Chinese medicine（TCM）in the treatment of POI in Vietnam.The pharmacodynamics mechanism of Yijing Decoction,an ancient prescription commonly used in lingnan Luo’s gynecological treatment of POI was studied through network pharmacology to discuss the possible mechanism of kidney-tonifying method.Methods:（1）Meta-analysis of kidney tonifying therapy for POI:The retrieval period was from The establishment of the databases to May 2021.Randomized controlled trials（RCTs）of kidney tonifying therapy for POI were retrieved from CNKI,Wanfang,VIP,Sino Med,Pub Med,Embase,and The Cochrane Library.The search terms were the combination of subject words and free words.Chinese search terms include premature ovarian insufficiency,primary ovarian insufficiency,tonifying the kidney,nourishing the kidney,Warming Yang,Chinese Medicine,Chinese Medicine,acupyncture,embedding,etc.The treatment group was treated with kidney tonifying therapy alone or added kidney tonifying therapy based on the control group.The control group was treated with conventional therapy or conventional therapy combined with single TCM treatment.In addition,the references of the included literatures were browsed,and the relevant literatures were manually retrieved and supplemented.Data were screened and extracted according to inclusion criteria and exclusion criteria.The quality of the literature was assessed using the risk of bias assessment recommended by the Cochrane Manual.Review Manager 5.3 software was used for meta-analysis.Relative risk（RR）was used as the effect index for measurement data,and mean difference（MD）was used as the effect index for enumeration data.I² test was used for heterogeneity analysis,and subgroup analysis,sensitivity analysis and other methods were used to analyze the sources of heterogeneity.Funnel plot or EGGER analysis was used to determine whether the results of a single outcome indicator were biased.（2）Lingnan Luo’s gynecology on drug rules in the treatment of POI:Clinical cases of POI treated by the third generation inheritor Luo Songping and the fourth generation inheritor Zhu Ling of Lingnan Luo’s Gynecology were collected.From January 2009 to December 2019,the clinic was located at the Department of Gynecology of the First Affiliated Hospital of Guangzhou University of Chinese Medicine.The patient’s general basic information,including history of present illness,relevant examinations,TCM pattern and prescriptions was extracted.SPSS 25.0 and SPSS Modeler 18.0 were used to analyze the valid survey data,and descriptive statistics were conducted on the clinical characteristics and risk factors of POI.Data mining methods,such as frequency analysis,association rule analysis and system cluster analysis,were used to analyze the regularity of drugs.According to high-frequency drugs use,association rules and systematic cluster analysis.（3）Pharmacology of Yijing Decoction network in the treatment of POI:With bioavailability and medicinal properties as screening conditions,the active ingredients of Yijing Decoction were collected from TCMSP,and the target information of effective active compounds was collected from STRING database and Uniprot database.Disease targets for POI were collected from the Gene Cards database.The identical targets of disease targets and Yijing Decoction action targets were taken as data sets and imported into Cytoscape 3.7.2 to construct a network of“target-active ingredient-Yijing Decoction drugs”.The identical targets also were imported into STRING database to construct protein-protein interaction network.According to the degree value result of STRING,the TOP 20 genes were taken as core genes and imported into R software to enrich biological process,cell composition,molecular function and signaling pathway by using the Cluster Profiler package and GGplot2.Results:（1）Meta-analysis of kidney tonifying therapy in the treatment of POI:A total of 38literatures were included,including 1282 patients in the experimental group and 1278patients in the control group.All of the included studies referred to"randomization",and 22 of them described specific randomization methods.Two studies used blind.One article mentioned lost to follow-up data processing.The total effective rate of kidney tonifying method was better than that of the control group,with statistical significance（RR=1.21,95%CI[1.16,1.27],P<0.00001）.There was no statistical difference in the incidence of adverse reactions between the two groups（RR=0.65,95%CI[0.41,1.02],P=0.06）.After treatment,FSH（SMD=-1.03,95%CI[-1.43,-0.63],P<0.00001）and LH（SMD=-0.77,95%CI[-1.13,-0.41],P<0.00001）in treatment group using Chinese medicine were significantly lower than those in control group.AND AMH（SMD=1.12,95%CI[0.53,1.70],P=0.0002）,E₂（SMD=1.04,95%CI[0.6,1.48],P<0.00001）,AFC（SMD=0.86,95%CI[0.59,1.13],P<0.00001）in treatment group using Chinese medicine were significantly higher than those in control group.There was no statistical difference in the regulation of endometrial thickness between two groups（SMD=0.2,95%CI[-0.24,0.63],P=0.38）.After treatment,FSH（SMD=-0.25,95%CI[-0.85,0.35],P=0.41）,LH（SMD=-0.31,95%CI[-0.77,0.14],P=0.17）,E₂（SMD=0.41,95%CI[-0.01,0.84],P=0.06）,AMH（SMD=0.61,95%CI[-0.03,1.25],P=0.06）and AFC（SMD=-0.48,95%CI[-0.99,0.04],P=0.07）in treatment group using acupuncture were not statistically different from control group.The endometrial thickness（SMD=1.48,95%CI[0.92,2.05],P<0.00001）was significant thicker than that of the control group.After treatment,FSH（SMD=-0.80,95%CI[-1.13,-0.47],P<0.00001）in treatment group using acupuncture combined Chinese medicine was significantly lower than that in control group.E₂（SMD=0.89,95%CI[0.46,1.32],P<0.00001）,AFC（SMD=0.80,95%CI[0.51,1.10],P<0.00001）and endometrial thickness（SMD=0.96,95%CI[0.30,1.02],P=0.004）were significantly higher than those in the control group.There was no significant difference in LH regulation between the control group and the control group（SMD=-0.14,95%CI[-1.06,0.78],P=0.77）.（2）Lingnan Luo’s gynecology on drug rules in the treatment of POI:A total of 112patients were enrolled,characterized with the age ranging from 17 to 39 and the menophania age ranging 11 to 19.Abnormal menstrual cycle,menstrual period and menstrual volume occurred in POI,mainly manifested as decreased menstrual volume,late menstrual period and amenorrhea.Number of births in POI patients:Among the patients with POI,the number of patients with none pregnancy or none deliveries were accounted for the largest proportion.752 precreptions were collected.The analysis of traditional Chinese medicine（TCM）syndromes of POI and clinical symptoms:Patients were manifested as tidal fever,night sweats,sleep disorders,emotional depression,decreased quality of sexual life,palpitations and pain during sexual intercourse.In the first diagnosis 58 cases of Yin deficiency of liver and kidney,kidney deficiency and blood stasis,liver stagnation and kidney deficiency.The common drugs includes Cuscutae Semen,Radix Salviae Miltiorrhizae,Rehmanniae Radix praeparata,etc.High-frequency drugs mainly belong to the liver,spleen and kidney channels.The main drug groups include Chinese yam-Cuscutae Semen-Lycii Fructus,Corni Fructus-Codonopsis Radix-Cuscutae Semen.Cluster analysis showed that Corni Fructus,Cuscutae Semen and Herba epimedii were commonly used as Yang tonic drugs;Ligustrum lucidi and Lycii Fructus were commonly used as Yin tonic drugs;Bupleuri radix,Curcumae Radix and Cyperi rhizoma were commonly used as liver soothing and depression relieving drugs;Paeoniae radix rubra and Chuanxiong rhizoma and Achyranthis bidentatae radix were commonly used as blood activating and stasis removing drugs.（3）The Pharmacology of Yijing Decoction network in the treatment of POI:A total of 112 active ingredients of Yijing Decoction were obtained.There were 187identical targets of Yijing Decoction and POI.The protein-protein interaction（PPI）network was constructed by importing the targets of STRING,with a total of 187 nodes and 681 links.The PPI network was imported into Cytoscape 3.7.2 for topology analysis,and 159 potential targets were obtained according to Degree results.TP53,HSP90AA1,JUN,MAPK1 and AKT1 were potential targets of Yijing Decoction in the treatment of POI.R software analysis finally enriched 1299 GO items and 139pathways.Biological processes include regulation of regulation of DNA-binding transcription factor activity,respons to radiation,cellular response to oxidative stress,response to UV,etc.Cell components include transcription factors complex,vesicle lumen,and focal adhesive.Molecular functions include DNA-binding transcription factor binding,ubiquitin ligase binding,DNA-binding transcription activator binding,ATPase binding,etc.Signaling pathway enrichment included IL-17 signaling pathway,PI3K-Akt signaling pathway,TNF signaling pathway.Conclusions:（1）Meta analysis showed that kidney-tonifying therapy for POI can improve the total clinical response rate,adjust the hormone level in vivo,and promote the increase of the number of sinus follicles and the thickness of endometrium,while the incidence of adverse reactions is low.The herbs group had better regulating effect on sex hormones and the number of sinus follicles than the control group,while the acupuncture group had better regulating effect on endometrium than the control group.The regulation of sex hormones（FSH,E2）levels,AFC and endometrium thickness in the acupuncture and Chinese medicine group was better than that in the control group,suggesting that the combination of acupuncture and Chinese medicine may exert a complementary effect in POI treatment.（2）The basic treatment of POI in Lingnan Luo’s gynecology is tonifying kidney.The commonly used treatment includes regulating liver,activating blood circulation and invigorating spleen,etc.Commonly prescription include Zuogui Pill,Zuogui Yin,Guishen Pill,Luo’s Dingjing Decoction,Yijing Decoction,Luo’s Spleen&kidney Prescription,etc.Frequent drugs include Cuscutae Semen,Rehmanniae Radix Praeparata,Chinese yam,Spatholobi caulis,Curcumae Radix,Radix Salviae Miltiorrhizae.Main drug combinations included Chinese yam-Lycii Fructus-Cuscutae Semen,Spatholobi caulis-Curcumae Radix,Codonopsis Radix-Cuscutae Semen-Corni Fructus,Rehmanniae Radix Praeparata-Cuscutae Semen-Corni Fructus,Citri Reticulatae Pericarpium-Codonopsis Radix,etc.（3）The mechanism of Yijing Decoction in treating POI may be that ginsenoside,baicalin,quercetin and other active ingredients participate in il-17signaling pathway,PI3K-Akt signaling pathway and TNF signaling pathway through AKT,IL-6,Bcl-2 and other targets to regulate oxidative stress state and improve immune function and ovarian cell apoptosis in POI.Thus play the role of ovarian protection.