Mechanism of action of Wuzi Yanzong pill in the treatment of oligoasthenozoospermia in rats determined via serum metabolomics

Objective:To investigate the mechanism of action of Wuzi Yanzong pill(WYP)in rats with oligoasthenozoospermia(OAZ)via metabolomics and to provide a possible basis for improving this WYP-based treatment.Methods:A rat model of OAZ was established by treating male SpragueeDawley rats with glucosides from Tripterygium wilfordii Hook.F.Seventy-two rats were randomly divided into six groups:control,L-carnitine(positive control),model,and low-,medium-,and high-dose WYP groups.Rats in the experimental groups were treated with WYP for 4 weeks.At the end of the treatment period,sperm cell quality(density,motility,and viability)was assessed using a semen analysis system,mitochondrial membrane potential(MMP)was assessed using flow cytometry,and testicular injury was assessed using hematoxylin and eosin staining to validate the therapeutic effect of WYP in OAZ.Further,serum metabolomics-based analysis was performed using high-performance liquid chromatography-mass spectrometry to identify differential metabolic pathways and possible mechanisms of action of WYP in OAZ treatment.Results:A rat model of OAZ was considered successfully-established after comparing the quality of spermatozoa in the model group to that in the control group.WYP-M and WYP-H treatments significantly improved sperm cell density,motility,and viability compared with those in the model group(all P<.05).Compared with the model group,both WYP-M and WYP-H treatments increased MMP values(P=.006 and P=.021 respectively),while there was no significant difference in the L-carnitine group.L-carnitine and WYP administration reversed damage to the testes to varying degrees compared with that in the model group.Further,44 differential metabolites and four metabolic pathways,especially autophagy pathway,related to OAZ were identified via metabolomics.Conclusions:WYP improves sperm cell quality and MMP in OAZ primarily via autophagy regulation.These findings can be employed to improve the efficacy of WYP in humans.

Study on the mechanism of Wuzi Yanzong pill in treating osteoporosis based on network pharmacology and molecular docking

Background:Based on network pharmacology and molecular docking,our study discussed the mechanism of Wuzi Yanzong pill in treating Osteoporosis(OP),which lays the foundation for drug development of OP.Methods:The chemical compounds and potential targets of Wuzi Yanzong pill were explored through TCMSP,PubChem,Swiss ADME and other databases.GeneCards,OMIM and Drugbank databases were used to obtain OP related targets.The intersection between the targets of Wuzi Yanzong pill and the related targets of OP was found by drawing a Venn diagram.PPI network was constructed with the STRING database and core targets were screened.The TCM-compound-action target-disease network was drawn using the Cytoscape software.The Metascape platform was used to find the pathways and functions for core target enrichment.Molecular docking validation of action compounds and core targets is completed by software such as Auto Dock Vina.Results:59 compounds and 707 action targets of Wuzi Yanzong pill were found.603 disease targets were selected,106 intersection targets were found using a Venn diagram,and 37 core targets were screened.By enrichment analysis,143 KEGG pathways,1026 GO biological processes,23 GO cell compositions and 60 GO molecular functions were obtained.The results of molecular docking showed that the effective compounds of Wuzi Yanzong pill,such as stigmasterol,quercetin,kaempferol andβ-sitosterol,had high binding activity with STAT3,TNF and IL6 core target proteins.Conclusion:Wuzi Yanzong Pill may play a role in treating OP by regulating STAT3,TNF,IL-6,TP53,VEGFA,JUN,AKT1,IL-1B,SRC,MMP9 and other pathways,as well as cancer-correlation,rheumatoid arthritis-correlation,MAPK,Th17 cell differentiation,IL-17,TNF signaling pathway and so on,to interpret Wuzi Yanzong pill’s clinic.

Wuzi Yanzong prescription(五子衍宗丸)from Traditional Chinese Medicine for male infertility:a narrative review

Wuzi Yanzong prescription(WYP),as a classical prescription for male infertility with kidney essence deficiency,is composed of Gouqizi(Fructus Lycii),Tusizi(Semen Cuscutae),Wuweizi(Fructus Schisandrae Chinensis),Fupenzi(Fructus Rubi Chingii)and Cheqianzi(Semen Plantaginis).It has been used for hundreds of years in the treatment of male infertility,known as“the first prescription of ancient and modern seeds”,with convincing clinical evidence.At present,more than 100 chemical compounds have been isolated from WYP,including polysaccharide,fatty acids,flavonoids,phenylpropanoids,organic acids,alkaloids,terpenoids,etc.Pharmacological and clinical studies show that WYP has an obvious effect on reproductive system diseases,especially male infertility,which has a very wide application prospect.It also has effects on the nervous system,inhibiting liver injury,lowering blood sugar and blood lipid,anti-aging,improving immunity,resisting hypoxia and fatigue effects.This study reviewed the chemical constituents,quality control,pharmacology,and clinical application of WYP.There is no doubt about the clinical value of WYP,but its quality control system is not perfect,pharmacological mechanism is not fully explained,and clinical applications need to be reevaluated.Therefore,the follow-up researches should proceed from the theory of Traditional Chinese Medicine(TCM)and clinical applications,further explain the theoretical connotation,reveal the mechanism of action,and provide the basis for the secondary development of classic famous prescriptions.In addition,WYP is mostly used in combination with western medicines besides being used alone.Whether it can improve the efficacy and reduce side effects will also be a meaningful research direction in the future.

Wuzi Yanzong Pill Plays A Neuroprotective Role in Parkinson’s Disease Mice via Regulating Unfolded Protein Response Mediated by Endoplasmic Reticulum Stress

Objective:To investigate the protective effects and its possible mechanism of Wuzi Yanzong Pill(WYP) on Parkinson’s disease(PD) model mice.Methods:Thirty-six C57BL/6 male mice were randomly assigned to 3 groups including normal,PD,and PD+WYP groups,12 mice in each group.One week of intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP) was used to establish the classical PD model in mice.Meanwhile,mice in the PD+WYP group were administrated with 16 g/kg WYP,twice daily by gavage.After 14 days of administration,gait test,open field test and pole test were measured to evaluate the movement function.Tyrosine hydroxylase(TH) neurons in substantia nigra of midbrain and binding immunoglobulin heavy chain protein(GRP78) in striatum and cortex were observed by immunohistochemistry.The levels of TH,GRP78,p-PERK,p-elF2α,ATF4,p-IRE1α,XBP1,ATF6,CHOP,ASK1,p-JNK,Caspase-12,-9 and-3 in brain were detected by Western blot.Results:Compared with the PD group,WYP treatment ameliorated gait balance ability in PD mice(P<0.05).Similarly,WYP increased the total distance and average speed(P<0.05or P<0.01),reduced rest time and pole time(P<0.05).Moreover,WYP significantly increased TH positive cells(P<0.01).Immunofluorescence showed WYP attenuated the levels of GRP78 in striatum and cortex.Meanwhile,WYP treatment significantly decreased the protein expressions of GRP78,p-PERK,p-elF2α,ATF4,p-IRE1α,XBP1,CHOP,Caspase-12 and Caspase-9(P<0.05or P<0.01).Conclusions:WYP ameliorated motor symptoms and pathological lesion of PD mice,which may be related to the regulation of unfolded protein response-mediated signaling pathway and inhibiting the endoplasmic reticulum stress-mediated neuronal apoptosis pathway.

五子衍宗丸对黄体功能不全不孕(肾精亏虚)患者的治疗效果观察

目的探讨五子衍宗丸对黄体功能不全不孕(肾精亏虚)患者的治疗效果。方法选取衡水市第二人民医院2020年2月至2022年1月收治的120例黄体功能不全不孕(肾精亏虚)患者,采用随机数字表法分为观察组和对照组,各60例。对照组予以地屈孕酮治疗,观察组予以地屈孕酮联合五子衍宗丸治疗。比较两组治疗前后症状评分、疗效、治疗前后内分泌和代谢指标、安全性及妊娠成功率。结果治疗后两组主症、次症评分与总分均降低且观察均较低(P<0.05);观察组总有效率高于对照组(P<0.05);治疗后两组血清孕酮(P)、黄体生成素(LH)、卵泡刺激素(FSH)、雌二醇(E_(2))水平,以及高温相持续时间(HTPD)、高低温差(HLTD)、基础体温曲线高温相评分(HPS)均升高,且观察组均较高(P<0.05);两组移行天数均减少且观察组较少(P<0.05);两组不良反应发生率差异无统计学意义(P>0.05);观察组妊娠成功率高于对照组(P<0.05)。结论对黄体功能不全不孕(肾精亏虚)以五子衍宗丸联合地屈孕酮治疗可减轻症状、增强效果、改善内分泌及代谢状态,安全性高,且能显著提高妊娠成功率。

五子衍宗丸对雷公藤多苷致肾病综合征雄鼠生殖损伤的机制研究

目的:观察五子衍宗丸干预肾病综合征SD大鼠对生精细胞T型Ca^(2+)通道及顶体酶PPEF1活性的影响及作用机制。方法:按照随机数字表法将70只SD大鼠分为空白组(蒸馏水1 mL灌胃)、肾病模型组(蒸馏水1 mL灌胃)、肾病模型+雷公藤多苷组(雷公藤多苷0.04 mg·kg^(-1)·d^(-1)灌胃)、肾病模型+五子衍宗丸组(五子衍宗丸1.07 g·kg^(-1)·d^(-1)灌胃)、肾病模型+雷公藤多苷+低剂量五子衍宗丸组(雷公藤多苷0.04 mg·kg^(-1)·d^(-1)与五子衍宗丸0.54 g·kg^(-1)·d^(-1)同时灌胃)、肾病模型+雷公藤多苷+中剂量五子衍宗丸组(雷公藤多苷0.04 mg·kg^(-1)·d^(-1)与五子衍宗丸1.07 g·kg^(-1)·d^(-1)同时灌胃)、肾病模型+雷公藤多苷+高剂量五子衍宗丸组(雷公藤多苷0.04 mg·kg^(-1)·d^(-1)与五子衍宗丸2.14 g·kg^(-1)·d^(-1)同时灌胃)。通过鼠尾静脉注射阿霉素造成肾病综合征模型,除空白组外,其余各组于实验第1天和第8天分别鼠尾静脉注射阿霉素,第1天以4.0 mg/kg剂量注射、第8天以3.5 mg/kg剂量注射。实验各组连续灌胃干预30 d。采用膜片钳放大法记录生精细胞T型Ca^(2+)通道电流变化情况;ELISA法测定顶体酶PPEF1活性;ELISA法测定血清中促性腺激素释放激素(GnRH)、雌二醇(E2)、促卵泡激素(FSH)、睾酮(T)和黄体生成素(LH)水平;荧光标记法测定顶体反应发生率。结果:与空白组比较,肾病模型组生精细胞T型Ca^(2+)活性降低(P<0.01)、顶体酶PPEF1活性降低(P<0.05)、性激素GnRH、E2、T水平降低(P<0.05),性激素FSH、LH水平升高(P<0.05);与肾病模型组比较,肾病模型+雷公藤多苷组生精细胞T型Ca^(2+)活性降低(P<0.01)、顶体反应发生率降低(P<0.05),性激素T水平降低(P<0.05),性激素FSH水平增高(P<0.05);与肾病模型+雷公藤多苷组比较,肾病模型+雷公藤多苷+中剂量五子衍宗丸组T型Ca^(2+)活性升高、顶体酶PPEF1活性升高、顶体反应发生率升高、性激素GnRH、E2、T水平升高,FSH、LH水平降低(P<0.05);与肾病模型+雷公藤多苷+低剂量五子衍宗丸组比较,肾病模型+雷公藤多苷+中剂量五子衍宗丸组、肾病模型+雷公藤多苷+高剂量五子衍宗丸组T型Ca^(2+)活性升高、性激素T水平升高(P<0.05)。结论:五子衍宗丸可以提高生精细胞T型Ca^(2+)电流、顶体酶PPEF1活性,促进顶体反应的发生,五子衍宗丸可以防治雷公藤多苷在肾病综合征治疗中的致不育副作用。

加味五子衍宗丸联合地屈孕酮片对黄体功能不全不孕症患者的干预效果

目的加味五子衍宗丸联合地屈孕酮片在黄体功能不全(LPD)不孕症患者中的应用效果。方法选取2021年11月—2022年12月180例LPD不孕症,随机数字表法分为对照组和观察组,每组90例,对照组采取地屈孕酮片治疗,于此基础上观察组联合加味五子衍宗丸治疗。观察2组疗效、不良反应、妊娠率,比较治疗前后中医证候积分、高温相评分(HPS)、性激素[孕酮(P)、雌二醇(E_(2))]、子宫动脉血流阻力指数(RI)、搏动指数(PI)、子宫内膜厚度、Th1/Th2细胞因子[白细胞介素-4(IL-4)、白细胞介素-13(IL-13)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)]及子宫内膜容受性相关miRNA表达。结果观察组总有效率86.67%(78/90)高于对照组65.56%(59/90)(P<0.05)。治疗后观察组各中医证候积分、RI、PI、TNF-α、IFN-γ水平低于对照组,基础体温HPS、子宫内膜厚度、P、E_(2)、IL-4、IL-13水平高于对照组(P<0.05)。治疗后观察组miR-494-5p、miR-544b表达量低于对照组,miR-4427表达量高于对照组(P<0.05)。观察组不良反应总发生率10.00%与对照组6.67%比较,无显著差异(P>0.05);观察组妊娠率高于对照组(P<0.05)。结论在LPD不孕症患者治疗中,加味五子衍宗丸联合地屈孕酮片可增强疗效,改善子宫内膜容受性,促进胚胎植入,提高妊娠率,这可能与Th1/Th2免疫应答、miRNA调控有关。

五子衍宗丸对肾病综合征雄鼠生殖功能的保护作用

目的五子衍宗丸干预雷公藤多苷治疗肾病综合征,探究五子衍宗丸对肾病综合征大鼠生殖能力、肾脏功能的影响。方法以雄性SD大鼠为研究对象,按照随机数字表法将70只大鼠分为空白组、肾病模型组、肾病模型+雷公藤多苷组、肾病模型+五子衍宗丸组、肾病模型+雷公藤多苷+低剂量五子衍宗丸组、肾病模型+雷公藤多苷+中剂量五子衍宗丸组、肾病模型+雷公藤多苷+高剂量五子衍宗丸组。于实验第1天和第8天需造模各组分别鼠尾静脉注射阿霉素4.0、3.5 mg/kg造肾病综合征模型。从实验第1天开始实验各组连续灌胃干预30 d。采用全自动生化仪器检测血清尿素氮、血肌酐、24 h尿蛋白定量、总胆固醇、甘油三酯、血浆白蛋白;采用ELISA检测血清中GnRH、E_(2)、FSH、T和LH等激素水平。观察肾脏和睾丸的病理改变。结果睾丸病理结果显示与单独接受雷公藤多苷治疗的组别比较,加入五子衍宗丸的各组生精小管萎缩减轻,管腔空泡减少,各层生精细胞层数增多,生精细胞数目增多。肾脏病理结果显示联合应用雷公藤多苷和五子衍宗丸治疗的组别较单独应用雷公藤多苷或五子衍宗丸的组别肾小球硬化、肾小管损伤、肾间质纤维化和炎症细胞浸润等病理改变降低。与肾病模型组比较,肾病模型+雷公藤多苷组性激素T降低,FSH增高;与肾病模型+雷公藤多苷组比较,肾病模型+五子衍宗丸组性激素GnRH、E_(2)、T增高,FSH、LH降低;与肾病模型+雷公藤多苷组比较,肾病模型+雷公藤多苷+中剂量五子衍宗丸组、肾病模型+雷公藤多苷+高剂量五子衍宗丸组性激素GnRH、T增高,FSH、LH降低;与肾病模型+雷公藤多苷+低剂量五子衍宗丸组比较,肾病模型+雷公藤多苷+中剂量五子衍宗丸组、肾病模型+雷公藤多苷+高剂量五子衍宗丸组性激素T增高。与肾病模型组比较,肾病模型+雷公藤多苷+中剂量五子衍宗丸组、肾病模型+雷公藤多苷+高剂量五子衍宗丸组血清肌酐含量降低、血浆白蛋白含量升高。结论通过在雷公藤多苷治疗肾病综合征的基础上添加五子衍宗丸,观察到在肾脏和睾丸的病理、生化指标以及性激素水平方面,相较于未添加五子衍宗丸的组别,病变程度较轻。这表明五子衍宗丸能够有效治疗肾病综合征,并对男性不育问题产生积极影响。这一研究为同时处理肾病综合征和男性不育提供了新的治疗思路。

应用微阵列技术分析五子衍宗丸干预少弱精症小鼠睾丸组织的长链非编码RNA表达

目的研究五子衍宗丸干预少弱精症相关的睾丸长链非编码RNA(long noncoding RNA,lncRNA),初步探讨该方调控lncRNA在其干预少弱精症中的潜在作用机制。方法采用白消安法建立少弱精症动物模型,使用五子衍宗丸干预14天后,应用小动物精子自动检测仪测定附睾精子数量和活力及与雌鼠交配受孕率评估生育力。分别抽提正常组、模型组、药物组各6例睾丸组织的RNA,应用转录组测序技术分析并筛选五子衍宗丸调控的差异表达lncRNA,实时荧光定量(quantitative real-time PCR,qRT-PCR)技术对差异表达lncRNA数据验证。通过生物信息学对获得的药物调控lncRNA结合前期报道该方调控mRNAs的实测值,进行关联分析,建立该方调控lncRNA靶向mRNAs的共表达网络,筛选网络lncRNA靶向的核心mRNAs,对核心mRNAs进行KEGG信号通路分析,构建该方调控lncRNA靶向信号通路网络,分析五子衍宗丸调控lncRNA干预少弱精症的信号通路。结果五子衍宗丸可显著提高少弱精症精子质量、雌鼠受孕率,并明显调控296条睾丸lncRNA,其中上调215条,下调81条,qRT-PCR验证lncRNA测序结果可靠。通过lncRNA-mRNAs的关联分析筛选到药物调控的核心lncRNA(102条)和mRNAs(81条),该方调控lncRNA靶向信号通路网络显示,在102条lncRNA中,有80条lncRNA靶向2条及以上信号通路,信号通路中有40条对应2条及以上lncRNA,这些lncRNA靶向的信号通路涉及到睾丸细胞的生长、分化、代谢、凋亡、信号传导等整个细胞周期过程,并且主要富集在范科尼贫血通路、细胞周期、剪接体、叉头状转录因子通路、p53通路、同源重组等通路上。结论本研究提供了五子衍宗丸干预少弱精症的睾丸lncRNA表达谱,这些lncRNA参与了药物修复睾丸生精损伤,提示其可能在药效中起到了重要作用,是五子衍宗丸潜在的干预靶点。

肾茶精水方对雄性SD大鼠生殖功能的保护机制

目的:观察肾茶精水方对大鼠生殖功能的保护机制。方法:70只雄性SD大鼠随机分为7组,空白对照组、肾病综合征组(NS组)、肾病综合征+雷公藤多苷组(雷公藤组)、肾病综合征+雷公藤多苷+五子衍宗丸组(雷公藤+五子组)、肾病综合征+雷公藤多苷+低剂量肾茶精水方组(雷公藤+低剂量肾茶精水方组)、肾病综合征+雷公藤多苷+中剂量肾茶精水方组(雷公藤+中剂量肾茶精水方组)、肾病综合征+雷公藤多苷+高剂量肾茶精水方组(雷公藤+高剂量肾茶精水方组),每组10只。通过鼠尾注射阿霉素制备肾病综合征模型。灌胃干预30 d后取材。采用全自动生化仪检测UTP、Alb、BUN、TC、SCr、TG含量;采用ELISA法检测血清中GnRH、E2、FSH、T和LH激素水平;采用HE染色及Masson染色观察肾脏和睾丸的病理改变。结果:与雷公藤组相比,雷公藤+五子组、雷公藤+低、中、高剂量肾茶精水方组睾丸生精小管萎缩现象显著减轻,且生精细胞的数量和层数显著增加。肾脏病理状态也显著缓解,包括肾小球硬化程度降低、肾小管损伤减少、肾间质纤维化和炎症细胞浸润减轻。与NS组比较,雷公藤+五子组GnRH、E2、T、Alb增高,FSH、LH、BUN、SCr、TC、UTP、TG降低(P<0.05);雷公藤+中剂量肾茶精水方组FSH、BUN、SCr、TG水平降低,Alb含量升高(P<0.05),雷公藤+高剂量肾茶精水方组FSH、BUN、SCr、TC、UTP、TG含量降低,Alb含量升高(P<0.05);与雷公藤组比较,雷公藤+五子组GnRH、E2、T、Alb水平升高,FSH、LH水平降低(P<0.05),雷公藤+高剂量肾茶精水方组Alb水平增高(P<0.05)。结论:肾茶精水方可以减轻肾病综合征大鼠生化指标和病理结果的异常变化,肾茶精水方可以防治雷公藤多苷治疗过程中的致不育作用。

A Chinese Herbal Formula,Wuzi Yanzong Pill(五子衍宗丸),Improves Spermatogenesis by Modulating the Secretory Function of Sertoli Cells

Objective： To evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill （五子衍宗丸, WYP） on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism. Me^ods： Five groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group （5 in each group） were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f （GTW） either alone （20 mg/kg） or followed by WYP （0.5, 1.0, or 2.0 g/kg daily）, respectively for 30 days. Serum levels of follicle-stimulating hormone （FSH）, inhibin B （INHB） and testosterone （r） were evaluated using enzyme-linked immunosorbent assay. Androgen- binding protein （ABP） gene expression and transferrin （TF） protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen＇s score. Results： The toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen＇s scores were detected in the model group compared with the control group （P〈0.05 or P〈0.01）. Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW- induced changes in specific secretory functions of Sertoli cells （ABP, INHB and TF）. Furthermore, serum T level and Johnsen＇s score of the testis increased greatly compared with the model group （P〈0.01）. Conclusion： WYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.

Effect of Modified Wuzi Yanzong Granule(加味五子衍宗颗粒)on Patients with Mild Cognitive Impairment from Oxidative Damage Aspect

Objective： To observe the effects of modified Wuzi Yanzong Granule （加味五子衍宗颗粒,WYG） on memory function and the activity of serum superoxide dismutase （SOD）, malondialdehyde （MDA） levels, leukocyte mitochondrial DNA （mtDNA） deletion rate and β-amyloid protein1.28 （A β 1-28 ） in patients with mild cognitive impairment （MCl）. Methods： Thirty-six patients with MCl were selected based on the internationally recognized Petersen＇s criteria, and equally and randomly assigned to two groups. The treated group was treated with WYG and the control group was treated with placebo for 3 months. In addition, 20 healthy subjects were included in the study as the normal control group. Changes of memory function, SOD activity, MDA content, leukocyte mtDNA deletion rate and A β 1-28 content were observed before and after treatment. Results： Compared with the normal control group, the memory quotient and SOD activity in patients with MCl decreased significantly （P 〈 0.01）, while MDA, A β1-28 levels and the leukocyte mtDNA deletion rate increased significantly （P 〈 0.01）. After treatment, levels of memory quotient and serum SOD activity increased while the serum MDA level, leukocyte mtDNA deletion rate and A β 1-28 level decreased in the treated group compared with those before treatment （P〈0.01, P〈0.05）. Meanwhile, leukocyte mtDNA deletion rate and A β 1-28 content in the treated group were all lower than those in the control group （P〈0.05）. Conclusion： WYG could improve memory function in patients with MCl and the therapeutic mechanism is possibly related to the increased activity of anti-oxidase, the improved free radical metabolism and the alleviation of leukocyte mtDNA oxidation damage. WYG shows clinical significance in delaying the progression of MCl.

补肾方药治疗肾虚所致男性生殖功能衰减的药效和机制研究进展

有关高龄育子和少子化的现状表明,男性生殖功能衰减带来不育风险的防治问题日益突出。男性生殖功能衰减与下丘脑-垂体-性腺轴、氧自由基、血睾屏障、生精细胞凋亡等因素有密切关系。中医认为,肾藏精,主生殖,生殖功能衰减与肾之阴阳失衡密切相关,运用补肾法可治疗肾虚导致的生殖功能衰减。补肾类方药通过系统调节下丘脑-垂体-性腺轴的功能,可显著延缓生殖功能的衰退;补肾类方药可清除体内过多的氧自由基,保护细胞免受氧化损伤;补肾类方药可通过维持血睾屏障而改善生精功能,并能调控生精细胞的凋亡。通过补肾改善肾虚所致生殖功能衰减的代表性方剂有五子衍宗丸、肾气丸、右归丸、六味地黄丸、左归丸及龟鹿二仙膏。这些方剂并非只针对单一机制产生效果,而是通过多途径、多环节、多靶点发挥治疗作用,综合调节机体内环境,以达到改善男性生殖功能的目的。中医补肾疗法在男性生殖功能衰减的治疗中发挥着不可替代的作用。深入研究其作用机制,可更好地指导临床实践,为改善男性生殖功能衰减提供理论依据。

桂枝茯苓丸联合五子衍宗丸治疗精子顶体酶活性低下临床研究

目的探讨桂枝茯苓丸联合五子衍宗丸治疗精子顶体酶活性低下的临床效果。方法将2021年7月—2023年2月于东莞市中医院男科就诊的80例精子顶体酶活性低下患者,按照就诊顺序随机分为对照组(40例)和观察组(40例)。对照组给予左卡尼汀口服液、维生素E软胶囊治疗,观察组给予桂枝茯苓丸联合五子衍宗丸治疗,两组患者均治疗3个月。分别于治疗前后对两组患者精子浓度、前向运动精子百分率,以及顶体酶活性进行检测,根据精液的检测结果将观察组患者分为少精子症组(3例)、弱精子症组(20例)、正常组(17例)。对观察组和对照组患者临床疗效及中医证候主症、次症积分进行对比,将观察组和对照组患者治疗前后精液检测参数、顶体酶活性进行对比,将少精子症组、弱精子症组、正常组治疗前后精液检测参数、顶体酶活性进行对比。结果治疗后观察组临床总有效率为85.00%,高于对照组的65.00%(P<0.05);治疗后对照组和观察组患者中医证候主症、次症积分均下降,且观察组低于对照组(P<0.05);治疗前后两组患者精子浓度比较差异无统计学意义(P>0.05),治疗后观察组与对照组患者前向运动精子百分率与精子顶体酶活性定量均高于治疗前,且观察组高于对照组(均P<0.05);治疗后3组患者精子顶体酶活性定量、正常精子形态、精子活率均高于治疗前(均P<0.05),正常组患者前向运动精子百分率治疗前后比较差异无统计学意义(P>0.05),而少精子症组治疗后精子浓度高于治疗前(P<0.05),弱精子症组治疗后前向运动精子百分率高于治疗前(P<0.05),治疗后弱精子症组患者精子顶体酶活性定量高于少精子症组、正常组(均P<0.05)。结论桂枝茯苓丸联合五子衍宗丸应用于精子顶体酶活性低下患者的治疗中,可有效改善精子活力、形态,有助于精子顶体酶活性的提高,尤其是对于弱精子症患者的效果较为显著。

五子衍宗丸治疗男性少弱精子症的Meta分析

目的系统评价五子衍宗丸治疗少弱精子症的有效性。方法检索CNKI、Wanfang Database、VIP、CBM、Pubmed、Cochrane Library、Embase数据库中五子衍宗丸治疗少弱精子症的随机对照试验,检索时限为自建库-2022年7月31日,对文献进行筛选、资料提取,评估偏倚风险,采用RevMan 5.3软件进行Meta分析。结果最终共纳入16项研究,包含1960例患者,依据干预措施进行亚组分析,Meta分析结果显示,五子衍宗丸组治疗后在精子密度[MD=5.28,95%CI(1.55,9.02),P=0.006]、A级精子[MD=6.69,95%CI(5.01,8.38),P<0.00001]、A+B级精子[MD=10.57,95%CI(7.28,13.85),P<0.00001]、配偶妊娠率[RR=1.25,95%CI(1.13,1.39),P<0.00001]、总有效率[RR=1.46,95%CI(1.3,1.63),P<0.00001]均显著高于西药组;联合用药组治疗后在精液量[MD=0.88,95%CI(0.71,1.04),P<0.00001]、精子密度[MD=7.19,95%CI(1.92,12.46),P=0.007]、A级精子[MD=7.9,95%CI(4.59,11.2),P<0.00001]、A+B级精子[MD=8.52,95%CI(3.99,13.04),P=0.0002]、配偶妊娠率[RR=1.79,95%CI(1.31,2.44),P=0.0003]、总有效率[RR=1.23,95%CI(1.12,1.35),P<0.00001]均显著高于西药组。结论五子衍宗丸单用或联合西药在改善精子密度、A级精子、A+B级精子、配偶妊娠率、总有效率指标上优于西药。

加味五子衍宗丸治疗黄体功能不全不孕症患者的效果及对雌性激素、PAPP-A、VEGF的影响

目的分析加味五子衍宗丸治疗黄体功能不全(LPD)不孕症患者的效果及对雌性激素、妊娠相关蛋白(PAPP-A)、血管内皮生长因子(VEGF)的影响。方法选取2021年11月—2022年12月收治的160例LPD所致肾虚精亏证不孕症。采用随机数字表法分为对照组和研究组,各80例。对照组采用地屈孕酮片治疗,研究组在对照组基础上联用加味五子衍宗丸治疗,均治疗3个月经周期。比较2组临床疗效、妊娠成功率及治疗期间不良反应发生情况。比较治疗前后中医证候积分、血清PAPP-A、VEGF、卵泡素(FSH)、促黄体生成素(LH)、雌二醇(E_(2))和孕酮水平。结果研究组总有效率为93.75%(75/80)高于对照组的82.50%(66/80)(P<0.05)。治疗后,研究组FSH、LH、E_(2)、孕酮、PAPP-A、VEGF水平高于对照组(P<0.05)。随访3个月经周期,研究组妊娠成功率为51.25%(41/80)明显高于对照组的35.00%(28/80)(P<0.05)。2组治疗期间均无药物不良反应发生。结论地屈孕酮片联合加味五子衍宗丸治疗LPD所致肾虚精亏证不孕症获得较好临床效果,调节患者内分泌代谢,提高血清PAPP-A、VEGF水平,提高妊娠成功率。

五子衍宗丸治疗多囊卵巢综合征的网络药理学探讨

目的:基于中药网络药理学方法探讨五子衍宗丸治疗多囊卵巢综合征(Polycystic ovary syndrome,PCOS)的潜在作用机制。方法:通过检索TCMSP中苍附导痰丸组成药物的成分数据,检索Disgenet、GeneCards、OMIM数据库中PCOS疾病相关靶点;利用Biogenet分别对药物与疾病靶点进行蛋白质相互作用网络进行构建,并提取交集网络以获得五子衍宗丸治疗PCOS的关键靶点;利用Metascape对关键靶点进行GO和KEGG富集分析,得出五子衍宗丸治疗PCOS的作用机制。结论:五子衍宗丸治疗多囊卵巢综合征的主要成分可能是槲皮素、山奈酚、β-谷甾醇等,涉及到的关键通路有癌症信号通路、脂质与动脉粥样硬化信号通路、化学致癌–受体激活通路、T白细胞介素-17信号通路等,与激素反应、对有机物的反应、细胞对脂质的反应、对外源性刺激的反应等多个生物学过程相关。

五子衍宗汤联合桃红四物汤辅治2型糖尿病肾病肝肾阴虚兼血瘀证临床研究

目的:观察五子衍宗汤联合桃红四物汤辅治2型糖尿病肾病肝肾阴虚兼血瘀证的疗效。方法:64例随机分为观察组和对照组各32例。两组均予以西医基础治疗,观察组加用五子衍宗汤联合桃红四物汤。结果:观察组总有效率高于对照组(P<0.05)。两组治疗后中医症状积分、尿液指标、肾功能指标、肾动脉血流动力学指标及TNF-α、Fetuin-A、MCP-1指标均改善(P<0.05),且观察组改善优于对照组(P<0.05)。两组不良反应发生率比较差异无统计学意义(P>0.05)。结论:五子衍宗汤联合桃红四物汤辅治T2DN肝肾阴虚兼血瘀证可提高临床疗效,且安全。

Relationship between Tissue Distributions of Modified Wuzi Yanzong Prescription(加味五子衍宗方) in Rats and Meridian Tropism Theory

Objective： To investigate the relationship between tissue distributions of modified Wuzi Yanzong prescription（加味五子衍宗方, MWP） in rats and meridian tropism theory. Methods： A high-performance liquid chromatography with Fourier transform-mass spectrometry（HPLC-FT） method was used to identify the metabolites of MWP in different tissues of rats after continued oral administration of MWP for 7 days. The relationship between MWP and meridian tropism theory was studied according to the tissue distributions of the metabolites of MWP in rats and the relevant literature. Results： Nineteen metabolites, mainly flavanoid compounds, were detected in the different rat tissues and classified to each herb in MWP. Further, it was able to establish that the tissue distributions of the metabolites of MWP were consistent with the descriptions of meridian tropism of MWP available in literature, this result might be useful in clarifying the mechanism of MWP on meridian tropism. In the long run, these data might provide scientific evidence of the meridian tropism theory to further promote the reasonable, effective utilization, and modernization of Chinese medicine. Conclusion： The tissue distributions of MWP in vivo were consistent with the descriptions of meridian tropism of MWP.

五子衍宗丸对帕金森病小鼠神经元运动功能的作用及其可能的机制

目的观察五子衍宗丸(WYP)对帕金森病模型小鼠运动功能的作用及其可能的机制。方法24只雄性C57BL/6小鼠随机分为对照组、模型组和WYP组,每组8只。模型组及WYP组小鼠连续7 d腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶,建立帕金森病模型。WYP组小鼠从模型制备第1天开始,连续14 d采用WYP药液[16 g/(kg.d)]灌胃,2次/d;对照组和模型组同一时间用0.9%氯化钠溶液[50 ml/(kg.d)]灌胃,2次/d。步态实验检测各组小鼠行为学表现;免疫荧光染色检测各组小鼠黑质区酪氨酸羟化酶(TH)阳性细胞数,核因子E2相关因子2(Nrf2)的荧光强度和NeuN神经元与Nrf2a共染细胞数;Western blotting检测各组小鼠脑组织TH、Kelch样ECH相关蛋白1(Keap-1)、Nrf2、血红素加氧酶-1(HO-1)的表达水平。结果步态实验结果显示,与对照组比较,模型组小鼠左前爪、右前爪、左后爪、右后爪站立时间均明显缩短(P<0.01),左前爪、右前爪、左后爪、右后爪摆动时间均明显延长(P<0.05);与模型组比较,WYP组小鼠左前爪、右后爪站立时间延长(P<0.05),左前爪、右前爪摆动时间缩短(P<0.05)。免疫荧光染色和Western blotting检测结果显示,与对照组比较,模型组TH阳性细胞数与蛋白、Nrf2平均荧光强度与蛋白、HO-1水平降低(P<0.01),Keap-1水平增高(P<0.01),NeuN神经元与Nrf2共染细胞数降低(P<0.001);与模型组比较,WYP组小鼠脑组织TH阳性细胞数与蛋白、Nrf2平均荧光强度与蛋白、HO-1水平、NeuN神经元与Nrf2共染细胞数均增高(P<0.05),Keap-1蛋白水平降低(P<0.05)。结论五子衍宗丸可缓解帕金森病小鼠运动功能障碍,保护多巴胺能神经元,其作用机制可能与调控Keap-1/Nrf2/HO-1通路抑制氧化应激反应有关。