Erxian decoction potentially prevents postmenopausal osteoporosis by modulating miR-335: a study based on bioinformatics analysis and preliminary clinical case validation

Background:miRNAs are closely related to bone metabolism.Studies have shown that Erxian decoction can improve bone metabolism,possibly achieving this regulatory effect through miRNA targets.Netinfer was used to predict the miRNA targets of Erxian decoction for the treatment of postmenopausal osteoporosis,and the results were validated by clinical trials.Methods:In this study,we identified possible targets of Erxian decoction in osteoporosis by means of network pharmacological analysis and bioinformatic prediction.Fifteen cases of postmenopausal osteoporosis with kidney Yin and Yang deficiency(In traditional Chinese medicine,kidney Yin nourishes and moistens the tissues of the internal organs of the body,while kidney Yang promotes and warms the tissues of the internal organs of the body.)were treated with Erxian decoction for four weeks,and serum bone metabolism indices(P1NP,osteocalcin,andβ-CTX)and miRNA-335-5p expression were measured before and after treatment.Results:The constructed miRNA postmenopausal osteoporosis related gene network of the effective compound of the Erxian decoction has 296 points and 981 edges.The 39 postmenopausal osteoporosis related genes regulated by miRNA-335-5p were enriched in ossification,while the signaling pathways were enriched in rheumatoid arthritis,the Toll signaling pathway,the HIF-1 signaling pathway,and the MAPK signaling pathway.After taking Erxian decoction,the expression of the serum bone formation index(P1NP,osteocalcin)and miRNA-335-5p gene expression levels increased significantly.The alterations in P1NP and osteocalcin were correlated with the changes in miRNA-335-5p.Conclusion:Circulating miRNA-335-5p may serve as an important target of Erxian decoction in the treatment of postmenopausal women.The effect of Erxian decoction on bone formation is significant,but the underlying mechanism requires further investigation.

Observation on Therapeutic Effect of Erxian Decoction on Relieving Low Back Pain after PVP of PMOP-derived Vertebral Fracture

[Objectives]To explore the clinical effect of Erxian decoction on relieving low back pain after percutaneous vertebroplasty(PVP)of vertebral compression fracture caused by postmenopausal osteoporosis(PMOP).[Methods]Ninety patients who were treated in Suzhou TCM Hospital from September 2021 to January 2023 were randomly divided into three groups:traditional Chinese medicine group(n=30),western medicine group(n=30)and blank group(n=30).The patients in all the three groups were treated with basic anti-osteoporosis drugs.The patients in the traditional Chinese medicine group were treated with Erxian decoction after PVP,and those in the western medicine group were treated with celecoxib to relieve pain after operation.The visual analogue(VAS)score,Oswestry dysfunction index(ODI)score,TCM syndrome score and serum indexes such as interleukin-6(IL-6)and estrogen E2 were recorded before treatment and 2 weeks,1 month and 3 months after treatment.[Results](i)In terms of pain relief,the VAS score of the western medicine group was lower than that of the traditional Chinese medicine group after 2 weeks of treatment,but there was no significant difference in VAS score between the two groups after 1 month and 3 months,and the pain improvement of the two groups was better than that of the blank group.(ii)After 3 months of treatment,the ODI score in the traditional Chinese medicine group was lower than that in the western medicine group,and the improvement of TCM syndrome in the traditional Chinese medicine group was better than that in the other two groups 1 and 3 months after treatment(P<0.05).(iii)The level of IL-6 in the western medicine group was lower than that in the other groups after 2 weeks,and there was no significant difference between the two groups after 3 months of treatment.After 3 months of treatment,the level of E2 in the traditional Chinese medicine group was higher than that before treatment and higher than that in the western medicine group and the blank group,but there was no significant difference between the two groups(P>0.05).[Conclusions]Both Erxian decoction and non-steroidal anti-inflammatory drugs can relieve residual low back pain after PVP,and their long-term effects are similar,but Erxian decoction has more advantages in alleviating pain,muscle and joint pain and sensory abnormalities in postmenopausal women.Moreover,it is safe and reliable,and is worthy of clinical application.

Discussion on the mechanism of Erxian Decoction in the treatment of premature ovarian failure based on network pharmacology and molecular docking

Objective:To explore the mechanism of Erxian Decoction in the treatment of premature ovarian failure.Methods:Based on the method of network pharmacology and molecular docking,the active ingredients of each drug in Erxian Decoction were obtained by searching the TCMSP database;the premature ovarian failure disease targets were collected from the GeneCards,OMIM,PharmGkb and Drugbank databases,and the active ingredients and the disease gene targets were collected Click the intersection to get the predictive target of Erxian Decoction for the treatment of premature ovarian failure.Use Cytoscape 3.8.0 to construct a"drug-component-target-disease"network;construct a protein interaction network(PPI)and streamline the core network through STRING database and Cytoscape;use R Studio software to enrich the Erxiantang treatment POF with GO And KEGG pathway enrichment analysis.Use molecular docking technology to verify the results of the"drug-component-target-disease"network.Results:68 main active ingredients were screened,involving 182 gene targets,among which the main active ingredients include Quercetin,Luteolin,Kaempferol,etc.;The core target genes include RB1,TP53,FOS,CDKN1A,ESR1,AKT1,MAPK1,TNF,etc.;GO enrichment items were obtained,including the 2545 Biological Process(BP),89 Cellular Component(CC),212 Molecular Function(MF);KEGG pathways,including PI3K-Akt signaling pathway,MAPK signaling pathway,and AGE-RAGE signaling pathway in diabetic complications.The verification of the molecular docking results indicated that the main active ingredient has a good binding activity with the core target.Conclusion:This study preliminarily revealed that Erxian Decoction may play a role in the treatment of POF through multi-component,multi-target,and multi-channel synergy,which provides a reference for the next in-depth research.

Mechanism of Guilu Erxian Gum in the treatment of osteoporosis based on network pharmacology

Objective:This article uses the method of network pharmacology to study the related mechanism of Guilu Erxian Gum in the treatment of osteoporosis.Methods:Based on the TCMSP database,ETCM database,chemistry database,and DrugBank database,the potential active ingredients and related targets of Guilu Erxian Gum were obtained.The known therapeutic targets of osteoporosis were obtained from OMIM and Genecards databases,and the STRING database was used.A protein interaction network(PPI)of active ingredients-disease targets was established,and the topological parameters of the network were analyzed using Cytoscape 3.8.0 software to obtain key active ingredients and their targets.In R4.0.2,the Bioconductor data package was used to analyze the GO biological function and KEGG pathway analysis of key targets,and obtain the effective ingredients and targets of Guilu Erxian Gum for treating osteoporosis.Results:The prediction results show that Guilu Erxian Gum has 87 active ingredients and 2305 targets,and there are 4141 known therapeutic targets for osteoporosis.The two act together to obtain a total of 71 PPI core genes.The GO biological process analysis yielded 95 entries,and the KEGG pathway analysis yielded 115 pathways.Conclusion:The analysis results show that Guilu Erxian Gum may play an anti-osteoporosis effect by regulating inflammatory factors,promoting osteoblast differentiation,inhibiting osteoclast formation,and improving microcirculation.The pathways involved include TNF signaling pathways,IL-17 signaling pathway,NF-κB signaling pathway,HIF-1 signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,etc.This study provides a theoretical basis for further elucidating the pharmacological mechanism of Guilu Erxian Gum in the treatment of osteoporosis.

To explore the mechanism of Erxian decoction in treating insomnia based on network pharmacology

Objective:This paper aims to explore the mech-anism of Erxian Decoction in treating insoimnia by means of network pharmacology research method.Methods:The com-ponents and related targets of Erxian Decoction are screened by TCMSP database.Disease targets are obtained by OMMM and Gene Cards database,and the common targets of drugs and diseases are obtained.The network diagram of"drug-coumponent-disease-target"is constructed and analyzed.STRING database constructs PPI network and finds the core target.GO and KEGG enrichnent analysis are employed on intersection targets.Results:84 effective components and 169 drug targets.of Erxian Decoction as well as 2614 targets of insomnia are screened out.Seventy-two intersection targets are selected by Venn diagram,and the core targets include IL-6,TNF,VEGFA and L-1β.These intersection targets contain 404 GO processes and 67 KEGG pathways,including TOLL-like receptor signaling pathway,cyclic adenosine monophosphate(CAMP)signaling pathway and tumor necrosis factor(TNF)signaling pathway.Conclusion:Erxian Decoction may play a role in treating insomnia by regulating TOLL-like receptor signaling pathway,cyclic adenosine monophosphate(CAMP)signaling pathway and tumnor necrosis factor(TNF)signaling pathway.

龟鹿二仙胶诱导破骨细胞凋亡的机制研究

目的 探讨不同浓度龟鹿二仙胶对破骨细胞凋亡影响及相关机制。方法 RAW 264.7细胞培养传代,用M-CSF+RANKL诱导小鼠单核细胞RAW 264.7细胞株分化成破骨细胞,获得破骨细胞;CCK8法、TUNEL染色、流式细胞法检测不同浓度的龟鹿二仙胶对破骨细胞凋亡的影响;Real-time PCR法、Western blot法检测破骨细胞凋亡相关分子Bcl-2、Bax、Cytochrome C的表达;经龟鹿二仙胶及PI3K/AKT通路抑制剂LY294002干预后,通过流式细胞法、Western blot法检测破骨细胞凋亡率及凋亡相关蛋白AKT的变化。结果 不同浓度的龟鹿二仙胶(GLEXJ高、中、低)均能抑制破骨细胞的增殖活性,提高破骨细胞的凋亡率,提高凋亡相关分子Bax/Bcl-2的比值以及Cytochrome C的表达,其中,以中浓度GLEXJ作用72 h后影响最显著;经中浓度龟鹿二仙胶,以及PI3K/AKT通路抑制剂LY294002的干预,明显抑制了PI3K/AKT通路相关蛋白AKT蛋白的磷酸化,破骨细胞总凋亡率显著上升。结论 龟鹿二仙胶通过抑制PI3K/AKT通路活化作用而促进破骨细胞凋亡。

双侧卵巢切除大鼠下丘脑⁃垂体的转录组学特征及二仙汤的调节效应

目的利用RNA测序技术研究围绝经期综合征模型大鼠下丘脑-垂体的转录组学特征及二仙汤的作用。方法以大鼠双侧卵巢切除术模拟围绝经期综合征雌激素骤然下降状态。48只雌性SD大鼠,随机取假手术组12只,其余36只大鼠双侧卵巢切除术后再随机分为双侧卵巢切除组、17β-雌二醇组、二仙汤组,每组12只。大鼠双侧卵巢切除术后第7天,17β-雌二醇组灌胃17β-雌二醇(0.1 mg·kg^(-1)·d^(-1)),二仙汤组灌胃二仙汤提取液(8 g·kg^(-1)·d^(-1)),每日灌胃1次,持续6周;假手术组与双侧卵巢切除组均灌胃等量的灭菌水。采用酶联免疫吸附测定(ELISA)法检测血清雌二醇(E2)、促性腺激素释放激素(GnRH)与β-内啡肽(β-EP)含量。采用mRNA转录组测序技术检测下丘脑与垂体,根据FPKM值与edgeR算法筛选组间差异表达基因,进行基因本体(GO)与京都基因与基因组百科全书(KEGG)富集分析,并以实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法验证下丘脑雌激素受体α(Esr1)与线粒体编码的NADH脱氢酶亚基4(Mt-nd4)、垂体钙/钙调素依赖性蛋白激酶Ⅱα(Camk2a)与甲状腺激素受体α(Thra)共4个差异表达基因。结果①与假手术组比较,双侧卵巢切除组大鼠血清E2与β-EP显著下降(P<0.05);与双侧卵巢切除组比较,二仙汤组E2与β-EP水平显著升高(P<0.05)。②大鼠下丘脑与垂体可检测基因数量均为30957个。③与假手术组比较,双侧卵巢切除大鼠下丘脑上调基因135个、下调基因348个,垂体上调基因409个、下调基因82个。与双侧卵巢切除组比较,17β-雌二醇治疗后大鼠下丘脑上调基因126个、下调基因225个,垂体上调基因93个、下调基因273个;二仙汤治疗后大鼠下丘脑上调基因91个、下调基因92个,垂体上调基因390个、下调基因376个。④GO分析表明,下丘脑与垂体差异表达基因富集于细胞过程、生物学调控、代谢过程等。⑤KEGG呈现了下丘脑与垂体的差异表达基因富集前30个信号通路,其中二仙汤调节尼古丁成瘾、氮代谢、突触囊泡循环等信号通路。⑥RT-qPCR所验证的二仙汤显著调节的Esr1、Mt-nd4、Camk2a基因表达变化与转录测序结果一致。结论双侧卵巢切除大鼠下丘脑与垂体的神经内分泌系统处于紊乱状态,17β-雌二醇侧重于调节下丘脑基因,二仙汤对垂体调节更广、更深。

基于网络药理学的二仙汤“异病同治”乳腺增生和围绝经期综合征的作用机制

目的:基于网络药理学探讨二仙汤“异病同治”乳腺增生和围绝经期综合征的作用机制。方法:通过中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选二仙汤中6味中药的活性成分并预测其潜在靶点;通过人类基因数据库(the human gene database,GeneCards)及在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)平台检索乳腺增生和围绝经期综合征的疾病靶点,借助Venn在线工具构建药物与两种疾病的交集靶点。运用Cytoscape 3.7.2软件构建“药物-成分-靶点-疾病”网络;利用STRING数据库构建PPI网络,按Dgree值大于等于平均值进一步筛选核心靶点;采用DAVID数据库进行基因本体论(gene ontology,GO)富集分析,同时实现京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果 :根据口服生物利用度(oral bioavailability,OB)≥30%和药物类药性(druglikeness,DL)≥0.18筛选得到二仙汤有效活性成分104个,预测潜在作用靶点147个;通过筛选得到乳腺增生靶点4153个,围绝经期综合征靶点155个,药物与疾病的交集靶点29个,其中IL-6、VEGFA、TNF、TP53、ESR1、CAT、IL-1β等可能是二仙汤治疗乳腺增生和围绝经期综合征的关键靶点。活性成分及靶点主要参与DNA模板转录正调控、基因表达的正调控、胞外间隙、胞外区、类固醇结合、酶结合等;靶基因主要富集在TNF、NOD样体、HIF-1、癌症、PI3K-Akt等信号通路。结论:二仙汤可能通过调控PI3K-Akt、HIF-1等信号通路达到防治乳腺增生和围绝经期综合征的目的。

水陆二仙丹联合抵挡汤加减方对早中期糖尿病肾病患者的临床疗效

目的探讨水陆二仙丹联合抵挡汤加减方对早中期糖尿病肾病患者的临床疗效。方法83例患者随机分为对照组(42例)和观察组(41例),对照组给予厄贝沙坦片,观察组在对照组基础上加用水陆二仙丹联合抵挡汤加减方,疗程12周。检测临床疗效、中医证候评分、血糖指标(FBG、HbA1c)、血脂指标(TC、TG)、肾功能指标(BUN、Scr、24 h UTP、eGFR)、炎症因子(IL-1β、hs-CRP、IL-6、TNF-α、IL-18、TGF-β1)、免疫功能指标(淋巴细胞、中性粒细胞、CD8^(+)、CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+))、不良反应发生率变化。结果观察组总有效率高于对照组(P<0.05)。治疗后,观察组中医证候评分、血糖指标、血脂指标、BUN、Scr、24 h UTP、炎症因子、CD8^(+)降低(P<0.05),淋巴细胞、中性粒细胞减少(P<0.05),eGFR、CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)升高(P<0.05),并比对照组更明显(HbA1c、TG、SCr、24 h UTP、淋巴细胞、中性粒细胞除外)(P<0.05)。2组不良反应发生率比较,差异无统计学意义(P>0.05)。结论水陆二仙丹联合抵挡汤加减方可安全有效地改善早中期糖尿病肾病患者临床症状,其机制可能与降低炎症水平、改善机体免疫功能有关。

仝小林教授治疗帕金森病伴嗜睡验案一则

帕金森病是一种进行性神经退行性疾病,以肌强直、运动迟缓、静止性震颤等运动症状及便秘、认知、情绪、睡眠障碍等非运动症状为突出临床特征。帕金森病属中医学“颤证”“震颤”等范畴,仝小林教授综合“四焦八系”与“态靶理论”治疗帕金森伴嗜睡病例一则,提出颤证为髓系疾病,病位在顶焦,本案患者以“虚态”“老态”为本,痰浊、血瘀为标,以地黄饮子加减作为“调态方”联合“靶方”二仙汤加减治疗,取得良好临床疗效,为临床治疗帕金森病伴嗜睡提供了新的诊疗思路。

Systems Pharmacology Uncovers Multiple Mechanisms of Erxian Decoction (二仙汤) for Treatment of Premature Ovarian Failure

Objective:To predict the chemical compositions and drug targets and to systematically dissect the pharmacological mechanism of Erxian Decoction(二仙汤,EXD)as a treatment for premature ovarian failure(POF)using a systems pharmacology approach.Methods:The compounds present in EXD were obtained from three databases.The active ingredient was identified by analyzing the values of oral bioavailability(OB),drug-like ness(DL),and Lipin ski's rule(LR).The active in gredients were further searched in research articles,drug targets in the DrugBank database,and the C-T and T-P networks,as well as by pathway analysis using the Cytoscape platform.Results:A total of 728 compounds were identified in EXD.Of these,59 were identified as active compounds that conformed to the criteria with OB>30%and DL>0.18.By further searches in the literature,126 related targets were ide ntified that could in teract with the active compounds.Additi on ally,it was found that the beneficial effects of EXD in POF are probably exerted via regulation of the immline system,modulation of estrogen levels,and anti-oxidative activities,and that it may act in a synergistic or cooperative manner with other therapeutic agents.Conclusions:The systems pharmacology approach is a comprehensive system that was used to elucidate the pharmacological mechanism of EXD as a treatment for POF.The results of this study will also facilitate the application of traditional medicine in modern treatment strategies.

Guilu Erxian Glue(龟鹿二仙胶)Inhibits Chemotherapy-Induced Bone Marrow Hematopoietic Stem Cell Senescence in Mice May via p16INK4a-Rb Signaling Pathway

Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally(i.p.)with 5×10^6/mL H22 cells per mouse.Fifty tumor-bearing mice were divided into the control,model,pifithrin-α,GEG,and GEG+pifithrin-αgroups using a random number table,10 mice in each group.CTX(100 mg/kg i.p.)was administrated to mice from day 1 to day 3(d1–d3)continuously except for the control group.The mice in the pifithrin-α,GEG and GEG+pifithrin-αgroups were treated with pifithrin-α(2.2 mg/(kg·d)i.p.)for 6 consecutive days(d4–d9),GEG(9.5 g/(kg·d)i.p.)for 9 consecutive days(d1–d9),and GEG plus pifithrin-α,respectively.HSCs were collected after 9-d drug treatment.The anti-aging effect of GEG was studied by cell viability,cell cycle,andβ-galactosidase(β-gal)assays.The mRNA and protein expressions of cyclin-dependent kinase 2(CDK2),CDK4,inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16^(p16^INK4a),p21^Cip1/Waf1,p53,and phosphorylated retinoblastoma(pRb)were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot,respectively.Results Compared with the model group,GEG increased cell viability as well as proliferation(P<0.05 or P<0.01)and reducedβ-gal expression.Furthermore,GEG significantly decreased the expressions of p16^INK4a,p53 and p21^Cip1/Waf1 proteins,and increased the expressions of CDK2,CDK4 and pRb proteins compared with the model group(P<0.05 or P<0.01).Conclusion GEG can alleviate CTX-induced HSCs senescence in mice,and the p16^INK4a-Rb signaling pathway might be the underlying mechanism.

加味二仙汤对多囊卵巢综合征不孕患者胰岛素抵抗及性激素水平的影响

目的 探讨加味二仙汤对多囊卵巢综合征(Polycystic ovarysyndrome, PCOS)不孕患者胰岛素抵抗及性激素水平的影响。方法 选取2020年1月—2022年1月期间河南省濮阳市妇幼保健院收治的PCOS不孕患者120例,按随机数字表法分为对照组和观察组,每组各60例。对照组常规服用二甲双胍治疗,观察组在对照组基础上加用加味二仙汤治疗。1个月经周期为1个疗程,两组患者均治疗3个疗程。观察比较两组患者临床疗效、治疗前后胰岛素指标[空腹血糖(Fasting plasma glucose, FPG)、空腹胰岛素(Fasting insulin, FINS)以及稳态模式胰岛素抵抗指数(Homeostasis model insulin resistanceindex, HOMA-IR)]变化、性激素水平[雌二酮(Estradiol, E_(2))、促卵泡雌激素(Follicle stimulating hormone, FSH)、睾酮(Testosterone, T)以及促黄体生成素(Luteinizing hormone, LH)]变化、子宫及卵巢功能[卵巢体积、子宫内膜厚度、成熟卵泡数量]变化以及治疗后排卵、妊娠、复发情况。结果 治疗后观察组临床总有效率96.67%(58/60)高于对照组85.00%(51/60),差异有统计学意义(P<0.05)。治疗后两组患者胰岛素指标FPG、FINS、HOMA-IR均较治疗前明显降低,差异有统计学意义(P<0.05);且观察组胰岛素指标FPG、FINS、HOMA-IR均明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者性激素E_(2)、FSH、T、LH水平均较治疗前明显降低,差异有统计学意义(P<0.05);且观察组性激素E_(2)、FSH、T、LH水平均明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者卵巢体积较治疗前降低,子宫内膜厚度以及成熟卵泡数量较治疗前升高,差异有统计学意义(P<0.05);且观察组卵巢体积明显低于对照组,子宫内膜厚度以及成熟卵泡数量明显高于对照组,差异有统计学意义(P<0.05)。随访3个月经周期,观察组排卵率86.67%(52/60)、妊娠率76.67%(46/60)均高于对照组71.67%(43/60)、58.33%(35/60),复发率1.67%(1/60)低于对照组11.67%(7/60),差异有统计学意义(P<0.05)。结论 加味二仙汤治疗PCOS的临床疗效显著,可有效降低患者的胰岛素抵抗以及调节性激素水平,改善卵巢功能,促进排卵,提高妊娠成功率,值得临床推广。

基于HIF-1α/EGFR/MAPK1信号通路探讨龟鹿二仙胶对少弱精子症大鼠的保护作用

目的探讨龟鹿二仙胶调控缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)/表皮生长因子受体(epidermal growth factor receptor,EGFR)/和丝裂原活化蛋白激酶(mitogen-activated protein kinase 1,MAPK1)信号通路对少弱精子症(oligoasthenozoospermia,OAS)大鼠的保护作用。方法将48只SD雄性大鼠随机分为正常组,模型对照组,龟鹿二仙胶低剂量(0.65 g/kg)组、中剂量(1.25 g/kg)组、高剂量(2.5 g/kg)组和左卡尼汀组,每组8只。除正常组外,其他组均采用腺嘌呤灌胃给药进行造模。造模成功后进行灌胃给药进行干预,连续给药4周,记录大鼠一般情况,给药4周后进行取材处理。腹主动脉采血,ELISA法检测血清睾酮(testosterone,T)、雌二醇(estradiol,E2)、促卵泡激素(follicle-stimulating hormone,FSH)、促黄体生成激素(luteinizing hormone,LH)和催乳素(prolactin,PRL);分离肾组织,计算肾指数;分离右侧附睾,运用自动精子分析仪检测各组大鼠精子浓度和精子活力;HE染色观察睾丸组织病理改变;用试剂盒检测大鼠睾丸组织活性氧(reactive oxygen species,ROS)、超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平;免疫组化检测大鼠睾丸组织HIF-1α、EGFR和MAPK1的蛋白表达。结果与模型对照组相比,不同剂量的龟鹿二仙胶均可显著增加大鼠的体质量(P<0.01),降低大鼠的肾指数(P<0.01),提高精子浓度和精子活力(P<0.01),改善大鼠的精子质量,并能有效减轻模型大鼠睾丸组织损伤程度,其中与龟鹿二仙胶低剂量组相比,龟鹿二仙胶高剂量组大鼠体质量、精子密度和精子活力均显著上升(P<0.05)。ELISA结果显示,与模型对照组相比,不同剂量龟鹿二仙胶能显著升高T和E2水平(P<0.05),且显著降低FSH、LH和PRL水平(P<0.05)。ROS、SOD和GSH-Px检测结果显示,与模型对照组相比,龟鹿二仙胶低、中、高剂量组ROS水平显著降低(P<0.05),龟鹿二仙胶中、高剂量组和左卡尼汀组SOD和GSH-Px活性显著升高(P<0.05)。免疫组化结果显示,与模型对照组相比,龟鹿二仙胶中、高剂量组和左卡尼汀组大鼠睾丸组织HIF-1α、EGFR、MAPK1水平显著降低(P<0.05)。结论龟鹿二仙胶可能是通过HIF-1α/EGFR/MAPK1信号通路调节氧化应激来改善OAS。

二仙补骨活血汤联合芪骨胶囊对绝经后骨质疏松症患者骨代谢及预后的影响

目的:探讨绝经后骨质疏松症(PMOP)患者应用二仙补骨活血汤联合芪骨胶囊治疗的效果。方法:采用随机数字表法将2021年1月—2023年2月于南昌市洪都中医院就诊的80例PMOP患者分为两组,各40例。对照组口服碳酸钙D3+芪骨胶囊治疗,观察组加用二仙补骨活血汤治疗,两组均连续治疗6个月。比较两组临床疗效、骨代谢指标[β胶联降解产物(β-CTx)、Ⅰ型胶原氨基端延长肽(PⅠNP)]、骨密度(BMD)和血清性激素雌二醇(E_(2))、骨折率及安全性。结果:相比于对照组,观察组治疗总有效率更高,治疗后PⅠNP、β-CTx水平均更低,BMD和E_(2)水平均更高,差异均有统计学意义(P<0.05);两组骨折率比较,差异无统计学意义(P>0.05);两组均无明显不良反应发生。结论:PMOP患者应用二仙补骨活血汤联合芪骨胶囊治疗效果较佳,可改善骨代谢指标、BMD,提高E_(2)水平,且安全性好,利于预后。

加味二仙汤联合来曲唑对PCOS患者促排卵及血清性激素水平的影响

目的:探究加味二仙汤联合来曲唑治疗多囊卵巢综合征(PCOS)对患者促排卵及血清抗缪勒管激素(AMH)等性激素水平的影响。方法:选取本院2020年3月-2023年8月确诊的PCOS患者86例,随机数字表法分为西药组(n=43)和中西药组(n=43)。两组均服用来曲唑治疗,中西药组加服加味二仙汤治疗,统计两组治疗前后促排卵情况(成熟卵泡个数、宫颈粘液评分、排卵期子宫内膜厚度),性激素[雌激素(E_(2))、促卵泡激素(FSH)、睾酮(T)、黄体生成素(LH)]水平,AMH水平以及不良反应。结果:治疗后,中西药组成熟卵泡个数(5.3±0.6个)、宫颈粘液评分(11.9±1.4分)、排卵期子宫内膜厚度(9.2±1.1mm)、妊娠率(74.4%)均高于西药组(3.8±0.5个、9.2±1.1分、6.5±0.9mm、46.5%),两组血清E_(2)、T、LH、AMH水平均降低,FSH两组均升高,且中西药组变化幅度大于西药组(均P<0.05);两组不良反应发生率(16.3%、9.3%)无差异(P>0.05)。结论:加味二仙汤联合来曲唑治疗PCOS,可显著改善患者排卵情况及性激素水平,降低AMH水平,提高治疗妊娠率。

二仙汤治疗肾阳虚型老年性骨质疏松症临床观察

目的观察二仙汤治疗肾阳虚型老年性骨质疏松症的临床疗效。方法选取肾阳虚型老年性骨质疏松症患者90例,分为对照组与治疗组,各45例。对照组予口服碳酸钙D3片和阿仑膦酸钠,治疗组在此基础上加用二仙汤。比较2组治疗12周后临床疗效。结果治疗组总有效率高于对照组(P>0.05);治疗组VAS评分低于对照组(P<0.05);治疗后,2组骨密度水平差异无统计学意义(P>0.05);β-CTX治疗前后无明显变化,治疗组PINP、BGP高于对照组。结论二仙汤联合碳酸钙D3与阿仑膦酸钠治疗肾阳虚型老年性骨质疏松症疗效优于对照组,能延缓骨质疏松进展,缓解临床症状,可供临床参考使用。

Erxian Tang (二仙汤)——Introduction of a Chinese Herbal Formula, Clinical Practice, and Experimental Studies

Erxian Tang （二仙汤, EXT） is a Chinese herbal formula developed for the treatment of menopausal syndrome in women. In the past 50 years, EXT has shown positive efficacy in the treatment of many chronic diseases in TCM, involving syndrome types of Shen （肾） yin-yang deficiency, yin-deficiency caused yang-flourishing, and disharmony of Chong-Ren meridian. Experimental studies have revealed that EXT has multiple pharmacological actions on such multiple targets as hypothalamus-pituitary-target gland axis, immune function and free radical metabolism, etc.

二仙丸加减方含药血清抑制巨噬细胞焦亡的物质基础和作用机制

背景:课题组前期研究发现二仙丸加减方可以缓解胶原诱导性关节炎大鼠炎症反应,但其作用机制尚需进一步验证。目的:分析二仙丸加减方入血成分,观察二仙丸加减方含药血清对J774A.1巨噬细胞焦亡的影响。方法:①二仙丸加减方入血成分分析:采用超高效液相色谱-高分辨质谱(UHPLC-HRMS)对二仙丸加减方及入血成分进行检测和鉴定。②二仙丸加减方含药血清对J774A.1巨噬细胞焦亡的影响:采用分子对接技术对二仙丸加减方入血成分倍半萜类化合物与NLRP3进行初步验证。将J774A.1巨噬细胞随机分为空白对照组、脂多糖+三磷酸腺苷组及脂多糖+三磷酸腺苷+二仙丸加减方含药血清低(2.5%)、中(5%)、高(10%)剂量组。根据试剂盒说明书检测各组细胞上清液中乳酸脱氢酶释放情况;ELISA检测各组细胞上清液中白细胞介素1β、白细胞介素18水平;Hoechst/PI染色法检测各组细胞膜受损情况;Western blot检测各组细胞中NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达水平。结果与结论:①共鉴定出二仙丸加减方药效成分32个,入血成分21个,其中入血成分主要包括多种倍半萜类化合物;②分子对接结果显示3-O-Acetyl-13-deoxyphomenone、Incensol oxide、Atractylenolide Ⅲ、Rupestonic acid、3,7-Dihydroxy-9,11-eremophiladien-8-one与NLRP3之间结合活性较好;③与空白对照组相比,脂多糖+三磷酸腺苷组细胞上清中乳酸脱氢酶活性及白细胞介素1β、白细胞介素18水平均显著升高(P<0.001),Hoechst/PI染色可见PI阳性细胞数量显著增加。脂多糖+三磷酸腺苷+二仙丸加减方含药血清各组上述指标均显示不同程度降低;④与空白对照组相比,脂多糖+三磷酸腺苷组NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达显著升高(P<0.05);与脂多糖+三磷酸腺苷组相比,脂多糖+三磷酸腺苷+二仙丸加减方含药血清各组细胞中NLRP3、Caspase-1、GSDMD及GSDMD-N蛋白表达均有不同程度降低(P<0.05),且具有一定的剂量依赖性。结果表明:二仙丸加减方含药血清可能通过调控NLRP3/Caspase-1/GSDMD通路抑制J774A.1巨噬细胞焦亡。

二仙汤合桂枝茯苓丸加减治疗PCOS对患者代谢紊乱及卵巢血流动力学的改善作用

目的:观察二仙汤合桂枝茯苓丸加减治疗多囊卵巢综合征(PCOS)疗效及对患者代谢紊乱、卵巢血流动力学的影响。方法:将2021年3月-2023年3月本院就诊的PCOS患者104例分为两组,对照组(n=52)给予炔雌醇环丙孕酮片治疗,观察组(n=52)给予炔雌醇环丙孕酮片联合二仙汤合桂枝茯苓丸加减治疗,治疗3个月经周期后评估疗效。检测血清性激素及代谢指标,超声测量卵巢间质动脉收缩期峰值血流速度(PSV)、阻力指数(RI)和舒张末期血流速度(EDV)。结果:治疗后观察组中医证候积分(5.58±1.89分)、血清睾酮(2.01±0.33nmol/L)、空腹血糖(4.65±0.37mmol/L)、黄体生成素(7.12±0.64U/L)、总胆固醇(4.11±0.32mmol/L)、黄体生成素/卵泡刺激素(1.31±0.12)、低密度脂蛋白胆固醇(2.46±0.31mmol/L)、空腹胰岛素(11.07±2.65)、三酰甘油(1.09±0.19mmol/L)均低于对照组(7.96±2.03分、2.74±0.42nmol/L、5.22±0.51mmol/L、9.34±0.85U/L、4.52±0.36mmol/L、1.72±0.21、2.67±0.43mmol/L、12.89±2.97、1.23±0.21mmol/L),雌二醇(68.85±5.96ng/L)水平高于对照组(62.48±5.17ng/L)(均P<0.05),但两组卵泡刺激素和高密度脂蛋白胆固醇水平无差异(P>0.05),卵巢PSV(12.21±1.98cm/s)、EDV(10.02±2.17cm/s)均低于对照组(15.88±2.01cm/s、11.53±2.25cm/s),RI(0.82±0.21)和治疗总有效率(98.1%)均高于对照组(0.62±0.13、84.6%)(均P<0.05)。结论:二仙汤合桂枝茯苓丸加减治疗PCOS能够改善患者临床症状,纠正内分泌及糖脂代谢紊乱,改善卵巢血流动力学,提高临床疗效。