Systematic review of Yougui pills combined with levothyroxine sodium in the treatment of hypothyroidism

BACKGROUND Yougui pills have long been used to treat hypothyroidism,usually in combination with levothyroxine sodium in clinical treatment,while their clinical efficacy and safety are still controversial when compared to levothyroxine treatment alone.AIM To explore the clinical efficacy and safety of Yougui pills combined with levothyroxine sodium in the treatment of hypothyroidism.METHODS This meta-analysis was performed in accordance with the PRISMA guidelines.Randomized controlled trials on Yougui pills in the treatment of hypothyroidism published from 2008 to May 2021 were searched in a total of 8 databases(4 databases in Chinese and 4 databases in English).The quality of the included studies was evaluated according to the Cochrane risk assessment tool.Weighted mean difference(WMD)was used for continuous variables,and relative risk(RR)was used for binary variables.Data were extracted,and the meta-analysis was conducted with the statistical software of Stata15.0 and RevMan5.0.RESULTS A total of 140 articles were retrieved,and 9 of them were finally included,with a total sample size of 936 cases.The main meta-analysis results are as follows:(1)The group of Yougui pills combined with levothyroxine sodium had a significantly higher overall response rate than the group of levothyroxine sodium(RR=1.20,95%CI 1.12,1.28,P<0.00001);(2)Yougui pills combined with levothyroxine sodium achieved significantly better efficacy than levothyroxine sodium alone in alleviating adverse symptoms[standard mean difference(SMD)=-1.10,95%CI:-1.37,-0.84,P<0.00001];(3)The level of thyrotropin stimulating hormone in the group of Yougui pills combined with levothyroxine sodium was significantly lower than in the control group of levothyroxine sodium(WMD=-1.38,95%CI:-2.10,-0.67,P=0.00001);(4)The level of free triiodothyronine in the group of Yougui pills combined with levothyroxine sodium was higher than that in the control group of levothyroxine sodium(WMD=0.41,95%CI:0.03,0.79,P=0.03);(5)The level of free thyroxine in the group of Yougui pills combined with levothyroxine sodium was significantly higher than that in the control group of levothyroxine sodium(SMD=0.83,95%CI:0.44,1.22,P≤0.0001);and(6)The adverse reactions in the group of Yougui pills combined with levothyroxine sodium were significantly less than those in the control group of levothyroxine sodium(RR=0.33,95%CI:0.20,-0.53,P<0.00001).CONCLUSION In the treatment of hypothyroidism,the combination of Yougui pills with levothyroxine sodium may be better than levothyroxine sodium treatment alone.

Chinese herbal medicine Yougui Pill reduces exogenous glucocorticoid-induced apoptosis in anterior pituitary cells

Long-term glucocorticoid use may result in sustained suppression of one or more secreted components from the hypothalamo-pituitary-adrenal axis, and often results in apoptosis. Yougui Pill（YGP）, a 10-component traditional Chinese herbal medicine, has been shown to be clinically effective for glucocorticoid-induced suppression of the hypothalamo-pituitary-adrenal axis. However, the pharmacological and molecular mechanisms remain unclear. We hypothesized that YGP would exert an anti-apoptosis effect on dexamethasone-treated anterior pituitary cells. In vivo experiments showed that YGP significantly reduced the number of apoptotic cells, down-regulated m RNA expression of cytochrome c, caspase-3, and caspase-9, and up-regulated m RNA expression of Bcl-2. These findings suggest that YGP reduced glucocorticoid-induced apoptosis in rat anterior pituitary cells by regulating the mitochondria-mediated apoptosis pathway.

Network pharmacological study of Zuogui pill and Yougui pill for the treatment of postmenopausal osteoporosis

Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese medicine is also widely used for the treatment of PMO,especially in Asian countries.Zuogui pill(ZGP)and Yougui pill(YGP)are classical formulas for the treatment of PMO with potent clinical effects.This study aimed to explore the potential active compounds,potential targets,and potential mechanisms of ZGP and YGP for the treatment of PMO.Methods:Compounds from ZGP and YGP were collected from three online databases,and these compounds were screened by oral bioavailability and drug-likeness.Their corresponding targets were determined from the Medicine Systems Pharmacology Database and Analysis platform.The corresponding targets for PMO were obtained from two disease databases.The target protein-protein interaction was identified through the STRING platforms and the core targets were ascertained by analyzing the protein-protein interaction network by using Cytoscape software.PMO-related genes were identified via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses to identify specific biological processes,cellular components,molecular functions and key signalling pathways of ZGP and YGP for PMO treatment.Results:A total of 68 compounds were screened from ZGP with 168 putative potential target genes,whereas 99 compounds were screened from YGP with 214 potential target genes associated with PMO.Most of the core components included quercetin and kaempferol,and most of the core targets were TP53,AKT1,MAPK1,JUN,TNF,HSP90AA1,APP,IL6,VEGFA,RELA,MAPK8,NR3C1,EGFR,CXCL8,ESR1,FOS and MAPK14.Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that ZGP and YGP treat PMO primarily via regulation of bone formation and resorption,anti-inflammatory immunity and hormonal regulation.Conclusion:Our data provided insights into the mechanisms,by which ZGP and YGP treat PMO.This study points out a direction for further research into ZGP and YGP monomers and pathways and offers a new clinical treatment option for non-traditional Chinese medicine clinicians in the treatment of PMO.

Zuogui(左归丸)and Yougui pills(右归丸)improve perimenopausal syndrome via regulation of apoptosis in mice

OBJECTIVE:To examine,dissect,and understand the molecular mechanisms and combinatorial effects of Zuogui(左归丸,ZGP)and Yougui pills(右归丸,YGP)in 4-vinyl cyclohexene diepoxide(4-VCD)-induced Perimenopausal syndrome(PMS).METHODS:Using the 4-VCD-induced PMS mouse model,uterine and ovary index were measured,and serum sex steroidal hormone levels were evaluated after treatment with ZGP,YGP,ZGP+YGP,estradiol valerate(EV),and Gengnian An(更年安,GNA).Histopathological examinations,ingredient-target network predictions,western blotting,and real-time quantitative polymerase chain reaction(RT-q PCR)analyses were performed to ascertain the possible pharmacological effects and molecular mechanisms of ZYP and YGP.RESULTS:Treatment with ZGP and YGP remarkably improves estrous cyclicity and prevents pathological damage to the uterus.Also,altered sex hormones,including AMH,E2,FSH,LH,P,and T,were restored to normal levels after ZGP and YGP administration.Ingredient-target network analysis showed that the 5 ingredients common to the ZGP and YGP formula modulate 53 targets shared with PMS.Pathwayenrichment analysis further predicted that ZGY and YGP likely regulate of apoptosis and other essential pathways during PMS.In-vivo studies showed that ZGP and YGP suppress PMS via modulating apoptosis through decreasing Caspase-3 and Bcl-2-associated X(Bax)levels and increasing B-cell lymphoma-2(Bcl-2)/Bax and Bcl-2 levels.Importantly,ZGP+YGP treatment modulation effects were somewhat or significantly better compared to ZGP or YGP alone treatment.CONCLUSION:ZGP and YGP represent novel anti-PMS agents whose effects involve restoring altered hormonal levels,protecting the uterus,and regulating apoptosis.

基于网络药理学探讨右归丸干预强直性脊柱炎作用机制

目的基于网络药理学的研究方法探讨右归丸干预强直性脊柱炎的作用机制。方法通过中药系统药理学数据库与分析平台(TCMSP)检索并筛选出右归丸中药组成的有效化学成分及作用靶点信息,利用GeneCards及OMIM数据库获得强直性脊柱炎的靶点基因,通过VENNY 2.1.0获得右归丸组成中药与强直性脊柱炎的交集基因,借助String在线平台与Cytoscape 3.7.2软件获得PPI网络,在DAVID数据库完成GO及KEGG通路富集分析。结果共获得药物有效成分79个,有效基因靶点500个,疾病靶点基因767个,交集基因98个,PPI网络分析得到关键靶点基因是IL1B、IL6和TNF等。GO功能富集分析得到GOBP条目535条、GOMF条目88条和GOCC条目53条。KEGG富集分析得到121条通路,主要信号通路为IL-17、HIF-1和TNF等。结论该研究基于网络药理学理论,系统地分析了右归丸的中药组成可以通过多成分、多靶点、多通路等干预强直性脊柱炎的作用机制,进而起到调节免疫反应、调控炎症因子、保护软骨、延缓骨破坏等治疗作用。

右归丸辅助骨髓间充质干细胞对家兔骨质疏松性骨折愈合的影响及机制初探

目的探讨右归丸辅助骨髓间充质干细胞对家兔骨质疏松性骨折愈合的影响及可能机制。方法研究时间为2023年1月至6月。将50只家兔分为正常对照组、模型对照组、右归丸组、骨髓间充质干细胞组、联合治疗组,各10只。除正常对照组外其余各组建立骨质疏松性骨折模型,并予以相应药物治疗。治疗结束后,测定家兔股骨骨密度(bone mineral density,BMD)、骨小梁数量、骨小梁厚度、骨小梁分散度以及股骨弹性模量、刚度、最大应力、最大承载力;实时聚合酶链式反应及蛋白印迹法测定股骨Janus蛋白酪氨酸激酶2/信号转导和转录激活子3(Janus kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)基因与蛋白水平。采用单因素方差分析及LSD-t检验进行数据分析。结果模型对照组家兔股骨BMD、骨小梁数量、骨小梁厚度,股骨弹性模量、刚度、最大应力、最大承载力,股骨JAK2、STAT3 mRNA和蛋白水平均低于正常对照组(均P<0.05),骨小梁分散度高于正常对照组(P<0.05);右归丸组、骨髓间充质干细胞组、联合治疗组家兔股骨BMD、骨小梁数量、骨小梁厚度、股骨弹性模量、刚度、最大应力、最大承载力、股骨JAK2、STAT3 mRNA和蛋白水平均高于模型对照组(均P<0.05),骨小梁分散度低于模型对照组(均P<0.05);联合治疗组股骨BMD[(1.76±0.13)g/cm^(3)]、骨小梁数量[(5.87±0.24)个/mm]、骨小梁厚度[(0.14±0.03)mm],股骨弹性模量[(343.27±21.04)MPa]、刚度[(904.25±73.25)N/mm]、最大应力[(32.63±3.17)N/mm^(2)]、最大承载力[(358.85±13.02)N],股骨JAK2、STAT3 mRNA[(3.45±0.26)、(3.60±0.28)]和蛋白[(0.82±0.05)、(0.84±0.05)]水平高于右归丸组[(1.41±0.15)g/cm^(3)、(3.88±0.28)个/mm、(0.10±0.02)mm、(304.62±20.41)MPa、(698.52±69.55)N/mm、(25.34±3.22)N/mm^(2)、(297.34±13.24)N、(2.58±0.26)、(2.29±0.23)、(0.60±0.05)、(0.53±0.07)]、骨髓间充质干细胞组[(1.47±0.14)g/cm^(3)、(3.89±0.28)个/mm、(0.09±0.03)mm、(299.54±16.94)MPa、(720.33±69.48)N/mm、(23.24±3.13)N/mm^(2)、(289.38±13.13)N、(2.63±0.25)、(2.30±0.24)、(0.59±0.06)、(0.53±0.08)](均P<0.05),骨小梁分散度[(0.34±0.02)mm]低于右归丸组[(0.42±0.03)mm]、骨髓间充质干细胞组[(0.43±0.03)mm](均P<0.05)。结论右归丸辅助骨髓间充质干细胞能明显促进家兔骨质疏松性骨折愈合,其机制可能与右归丸辅助骨髓间充质干细胞治疗能激活骨质疏松性骨折家兔股骨JAK2/STAT3信号通路有关。

基于RAP1信号通路探讨右归丸延缓膝骨关节炎的软骨退变

目的通过右归丸治疗膝骨关节炎(KOA)模型大鼠,观察大鼠关节软骨形态及检测右归丸对RAP1信号通路表达和大鼠血清中的炎性因子表达,探讨右归丸对RAP1信号通路的表达及防治KOA的作用机制,为治疗提供理论基础。方法72只SPF级SD大鼠(雌雄各半)随机数字表法分为假手术组12只和造模组60只。造模组采用改良Hulth法建立膝关节OA模型,假手术组接受假手术。将造模成功的大鼠分为模型组(n=11)、右归丸低剂量组(n=12)、右归丸中剂量组(n=12)、右归丸高剂量组(n=12)及塞来昔布组(n=12)。给药阶段模型组、假手术组给予0.9%氯化钠溶液;右归丸低剂量组为等效浓度的1/2为0.5 g/d、右归丸中剂量组等效浓度为1 g/d、右归丸高剂量组为等效浓度2倍为2 g/d,塞来昔布组配置成悬浮(等效浓度为0.0072 g/d)。驱赶7周后检测并比较各组大鼠软骨病理,血清TNF-α、IL-1β和IL-18含量,以及大鼠软骨组织中RAP1、RAF、MEK1/2、ERK1/2、CREB、MMP3、MMP13、ADAMTS、C-fos、C-myc和C-Jun的表达水平。结果各组大鼠软骨病理检测结果显示:假手术组大鼠软骨组织未见异常,模型组大鼠关节软骨部分区域出现明显缺损,右归丸低剂量组大鼠关节软骨表面磨损较模型组有所减轻,而中、高剂量组及塞来昔布组大鼠软骨表面可见侵蚀现象明显改善。右归丸对KOA大鼠血清中的IL-1β、TNF-α、IL-18含量与模型组相比,右归丸高、中、低剂量组和塞来昔布组在大鼠血清中表达显著降低,差异具有统计学意义。与软骨组织中RAP1-RAF-MEKl/2-ERKl/2-CREB信号通路活化水平检测结果显示:与假手术组比较,模型组大鼠软骨组织中RAP1、RAF、MEK1/2、ERK1/2、CREB、MMP3、MMP13、ADAMTS、C-fos、C-myc和C-Jun的表达水平均显著升高,差异有显著统计学意义(P<0.01)。结果提示,在KOA发生时,RAP1-RAF-MEKl/2-ERKl/2-CREB信号转导通路激活,介导软骨组织中炎症反应及细胞凋亡的发生,这与以往研究结果一致。右归丸干预后,软骨组织中RAP1、RAF、MEK1/2、ERK1/2、CREB、MMP3、MMP13、ADAMTS、C-fos、C-myc和C-Jun蛋白表达水平均显著降低,差异有显著统计学意义(P<0.01)。结论右归丸能够抑制KOA大鼠软骨组织中RAP1-RAF-MEKl/2-ERKl/2-CREB信号通路的活化,降低炎性因子表达,从而延缓软骨退变。

右归丸对激素型肾阳虚证miRNA表达谱的影响及其靶基因功能分析

目的探讨微小RNA(miRNA)在右归丸对激素型肾阳虚证肾组织与正常肾组织中的表达差异,并用生物信息学方法分析其意义,为右归丸干预肾阳虚证的进一步机制研究奠定基础。方法15只SD大鼠随机分为正常组(5只)和造模组(10只),正常组注射2 mL生理盐水,造模组后肢肌注氢化可的松注射液。成模后将造模组随机分为模型组和右归丸组,每组5只。右归丸组给予右归丸汤剂灌胃,模型组和正常组给予生理盐水灌胃。麻醉处死后使用TRIzol法提取肾组织中RNA,使用NanoDrop ND-1000对总RNA进行质量检测,用标记后的RNA与Agilent Rat 8×60k miRNA芯片进行杂交。收集原始数据进行标准化处理,进行聚类分析和火山图分析。利用生物信息学方法,预测差异miRNA富集的基因本体论(GO)功能和京都基因与基因组百科全书(KEGG)通路。结果筛选出右归丸干预肾阳虚证相关差异表达miRNA 18条(|log 2FC|≥1.5,P≤0.05),其中包括5条上调miRNA,分别是rno-miR-149-3p、rno-miR-188-5p、rno-miR-221-5p、rno-miR-762、rno-miR-877;13条下调miRNA,分别是rno-miR-101b-5p、rno-miR-125a-3p、rno-miR-125b-2-3p、rno-miR-3085、rno-miR-344a-5p、rno-miR-347、rno-miR-34b-3p、rno-miR-351-3p、rno-miR-3573-3、rno-miR-370-3p、rno-miR-409a-3p、rno-miR-448-3p、rno-miR-503-3p。GO分析显示差异表达miRNA主要参与DNA、RNA转录的正调控进程且与ATP的结合显著相关。KEGG分析显示差异表达miRNA显著富集于cAMP通路和FoxO信号通路,这两条通路与线粒体功能密切相关。结论右归丸通过干预miRNA的表达,改善肾阳虚证大鼠线粒体功能,调节能量代谢异常,进一步为右归丸干预肾阳虚证的机制研究提供理论依据。

Research on the Mechanism of Zuogui Pill(左归丸)and Yougui Pill(右归丸)in Promoting Axonal Regeneration in Model Rats of Autoimmune Encephalomyelitis

Objective：To study the molecular mechanism of Zuogui Pill（左归丸,ZGP） and Yougui Pill（右归丸,YGP） on axonal regeneration in rats with experimental autoimmune encephalomyelitis（EAE）.Methods： EAE rat model was established by bilateral rear pedes subcutaneous injection of antigen made by mixing myelin basic protein（MBP） and complete Freud＇s adjuvant（CFA） in the volume ratio of 1：1.The pathological changes of axonal injury and regeneration in the brain and the spinal cord were observed on the 14th（the acute stage） and the 28th day（the remission stage） after modeling,with hematoxylin-eosin（HE） staining,silver stain, and immunohistochemical staining.The rats treated with prednisone acetate were taken as controls.Results： Observation under the light microscope with HE staining showed a sleeve-like change in rats＇ cerebrospinal parenchyma with inflammatory cell infiltration around the small vessels and neuronic denaturation,while silver staining showed excessive tumefaction and abscission of axon,and immunohistochemical analysis showed decreasing of nerve growth factor（NGF） expression at the acute stage of EAE,which was even more remarkable at the remission stage,showing significant difference as compared with the normal control（P〈0.05）. And the expressions of Nogo A,an axon growth inhibitor,and its receptor（Nogo-66 receptor,Ng R） were significantly higher than those in the normal control at the acute stage（P〈0.01）.However,after the intervention of ZGP and YGP,the pathological changes and axon damage in rats＇ brain and spinal cord were much more alleviated,and the NGF expression was significantly higher than that in the model group at the acute stage （P〈0.05）.The expression of NGF was even stronger during the remission stage,and a better effect was shown by YGP.As for Nogo A and Ng R expressions,they were significantly lower than those in the model group at the acute stage（P〈0.05）,but a better effect was shown by ZGP.Conclusions：ZGP and YGP can prevent axonal injury and promote the axonal regeneration in rats of EAE,and the possible mechanism is to increase the expression of NGF and reduce the expression of Nogo A and its receptor.However,some differences are observed between the two Chinese preparations in their acting times and points,which provides a certain basis for revealing the modern connotation of the Chinese medicine theory on tonifying Shen（肾）-yin and Shen-yang.

Effects of Zuogui Pill(左归丸) and Yougui Pill(右归丸) on the Expression of Brain-Derived Neurotrophic Factor and Cyclic Adenosine Monophosphate/Protein Kinase A Signaling Transduction Pathways of Axonal Regeneration in Model Rats with Experimental Autoimm

Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA）signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis（EAE）,in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods：The rats were randomly divided into normal control（NC）,model（MO）,prednisone acetate（PA）,ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein（MBP）68-86.The brain and spinal cord were harvested on the 14th and 28th day postimmunization（PI）,the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction（PCR）,and the cAMP levels were detected by using enzyme-immunoassay method.Results：（1）On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group（P〈0.05 or P〈0.01）.（2）On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group（P〈0.05 or P〈0.01）,and the cAMP level in YGP group was higher than that in ZGP group（P〈0.05）.The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively（P〈0.01）.（3）On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively（P〈0.05）.（4）On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions：The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.

病证结合骨质疏松模型大鼠骨骼的生物力学性能分析

背景:肾虚是骨质疏松症的最主要病机,对肾阳虚、肾阴虚与骨质疏松的关系研究有利于病证结合临床诊疗模式的发展。目的:评价右归丸对肾阳虚、肾阴虚大鼠股骨生物力学性能的影响差异,从生物力学角度论证骨质疏松症病证结合、因证施药的科学性。方法:手术摘除60只雌性SD大鼠双侧卵巢建立骨质疏松模型,10周后将大鼠随机分为肾阴虚组(n=30)和肾阳虚组(n=30),肾阳虚组臀部肌肉注射氢化可的松注射液,肾阴虚组灌胃甲状腺片混悬药液,1次/d,连续14 d。造模结束后,每组取20只灌胃右归丸混悬液,1次/d,连续12周,剩余10只作为对照不灌胃。灌胃结束后使用Micro-CT扫描测量股骨微结构参数,在力学试验机上对股骨分别进行三点弯曲、有限元仿真、股骨头压缩及表面压痕分布实验。结果与结论:①Micro-CT扫描显示,与肾阴虚组比较,肾阴虚+右归丸组股骨骨密度、骨体积分数、骨表面密度、骨小梁数量、骨小梁分离度升高(P<0.05);与肾阳虚组比较,肾阳虚+右归丸组股骨骨体积分数、骨表面密度、骨小梁数量及骨小梁厚度升高(P<0.05);②三点弯曲实验显示,与肾阳虚组相比,肾阳虚+右归丸组股骨弹性模量、最大抗弯强度和弯曲断裂强度均降低(P<0.05),韧性提高(P<0.05);③有限元仿真显示,右归丸能显著提高肾阳虚组大鼠股骨抗骨折能力,对肾阴虚组大鼠不产生明显作用;股骨头压缩实验显示,右归丸可增强肾阳虚组大鼠股骨头抵抗形变的能力,但右归丸对肾阴虚组和肾阳虚组大鼠股骨头表面性能的影响无显著区别;④结果表明,右归丸可增强肾阳虚骨质疏松骨骼生物力学性能,但对肾阴虚骨质疏松骨骼无显著影响。

右归丸治疗腰椎间盘突出症:网络药理学分析活性成分及潜在治疗靶点

背景:右归丸具有温肾助阳、益精填髓的作用,临床多用于治疗肾阳虚证的腰椎间盘突出症且具有较好的疗效。目的:采用网络药理学、分子对接技术并结合动物实验探讨右归丸治疗腰椎间盘突出症的潜在靶点及作用机制。方法:①网络药理学分析:通过TCMSP等数据库获取经过筛选后的右归丸有效成分与作用基因靶点,在GeneCards等数据库中收集与腰椎间盘突出症相关的基因,取二者交集进行拓扑结构分析,得到主要活性成分及核心治疗靶点。使用R软件对核心治疗靶点进行GO功能及KEGG通路富集分析。②分子对接:运用Autodock和Pymol软件进行中药活性成分与核心治疗靶点分子结合能预测。③动物实验:将18只SD大鼠随机分为对照组、退变组与退变+右归丸组,每组6只。取退变组与退变+右归丸组大鼠,采用纤维穿刺法制备椎间盘退行性变模型,造模后2周,退变+右归丸组大鼠给予右归丸汤剂(1次/d)灌胃,连续给药2周。给药结束后,采用ELISA法检测血清中肿瘤坏死因子α质量浓度,苏木精-伊红染色后观察椎间盘纤维环及髓核细胞的形态学改变。结果与结论:①筛选出右归丸有90种活性成分和64种靶点,且主要活性成分为槲皮素、山奈酚、β-胡萝卜素、大豆黄素,4′-O-甲基尼亚萨酚。右归丸治疗腰椎间盘突出症的核心靶点为白细胞介素6、肿瘤坏死因子α、AKT1、白细胞介素1B、血管内皮生长因子A。富集分析发现,交集基因可能通过白细胞介素17、肿瘤坏死因子、MAPK、PI3K-AKT等信号通路改善椎间盘退变。②分子对接实验验证了右归丸中的槲皮素、山奈酚、β-胡萝卜素等与核心靶点有较强的结合能力。③动物实验结果显示,退变组大鼠血清肿瘤坏死因子α质量浓度高于对照组(P<0.05),退变+右归丸组大鼠血清肿瘤坏死因子α质量浓度低于退变组(P<0.05);苏木精-伊红染色显示,退变组大鼠椎间盘纤维环、髓核结构遭到破坏,髓核细胞数量减少;退变+右归丸组大鼠椎间盘纤维环可见重建趋势,髓核细胞数量较退变组增加。④结果表明,右归丸可能通过槲皮素、山奈酚、β-胡萝卜素等主要活性成分作用于肿瘤坏死因子α等核心靶点改善椎间盘退变,进而治疗腰椎间盘突出症。

温通与温补理论探析

温法有温通与温补之别,温通是温以促通,温补是温以达补。首先,从三个代表性时期论述温法中“通”与“补”的历史脉络:仲景时期,治阳重在温通,甘草干姜汤演化为四逆汤类方,温通思想贯穿始终;金元时期,李东垣在升降思想下创制补中益气汤,其以象春生夏长之令以升补清阳;明清时期,温补学派在东垣脾胃学说的基础上,进一步强调了温阳补虚基础上填补阴精的重要性。其次,基于《伤寒论》太阳病篇、张景岳右归丸,分别探讨温通与温补的不同特点,同时从病因、病位、病机、药物组成等方面阐述温通与温补的应用鉴别。最后,从胸痹与久泻举例论证温通与温补的不同应用场景。

左、右归丸对去卵巢大鼠骨髓单核/巨噬细胞破骨分化的影响

目的基于AMPK/mTOR信号通路研究左、右归丸对去卵巢大鼠骨髓单核/巨噬细胞(bone marrow-derived macrophage,BMMs)破骨分化的作用及机制。方法将60只雌性大鼠随机分为6组,分别为空白组(KB)、假手术组(SHAM)、模型组(OVX)、补佳乐组(BJL)、右归丸组(YGW)和左归丸组(ZGW)。KB组不做处置,SHAM组模拟卵巢切除手术过程,后4组均将双侧卵巢切除。各组分别灌胃蒸馏水(SHAM、OVX)、补佳乐(BJL)、右归丸(YGW)和左归丸(ZGW)。12周后取大鼠股骨和胫骨骨髓,用差时贴壁法获得大鼠BMMs并进行破骨分化诱导。用抗酒石酸酸性磷酸酶活性检测试剂盒检测TRAP活性;qrt-PCR法检测TRAP基因的表达;Western blotting法检测TRAP、RANKL、AMPKɑ、p-AMPKɑ、mTORC1、p-mTORC1蛋白的表达。结果左、右归丸能显著降低去卵巢大鼠BMMs破骨诱导后TRAP活性,降低TRAP基因和蛋白的表达,降低与RANK结合的RANKL蛋白的表达。与KB组比较,OVX组AMPKɑ蛋白磷酸化水平降低,mTORC1蛋白磷酸化水平升高;3个用药组皆能对抗上述改变,其中左归丸优于右归丸。结论左、右归丸均能通过AMPK/mTOR信号通路抑制去卵巢大鼠BMMs的破骨分化。

右归丸联合丝裂霉素C对经尿道前列腺电切术治疗良性前列腺增生患者术后性功能和尿流动力学的影响

目的探讨右归丸联合丝裂霉素C(MMC)对经尿道前列腺电切术(TURP)治疗良性前列腺增生(BPH)患者术后性功能和尿流动力学的影响。方法选取2018年6月至2021年6月西安交通大学第二附属医院诊治的88例行TURP治疗的BPH患者作为研究对象。采用随机数字表法分为观察组和对照组,各44例。对照组术后给予MMC治疗,观察组术后给予MMC联合右归丸治疗,均持续治疗8个月。比较两组治疗前后中医证候积分、国际前列腺症状评分法(IPSS)评分、国际勃起功能障碍指数-5(IIEF-5)评分、生活质量(QoL)评分,以及男性性功能问卷(MSF-4)评分变化,比较两组治疗前后尿流动力学指标[最大尿流量(Q max)、最大逼尿肌压力(MDP)、强烈尿感容量(SD)、剩余尿量(PVR)、膀胱顺应性(BC)]变化,统计两组手术并发症发生情况。结果治疗后,观察组中医证候积分、IPSS、QoL评分低于对照组,IIEF-5、MSF-4评分高于对照组,PVR水平低于对照组,Q max、SD、MDP、BC水平高于对照组,差异具有统计意义(P<0.05)。两组并发症发生率比较,差异无统计学意义(P>0.05)。结论右归丸联合MMC应用于BPH患者TURP术后治疗,可显著改善患者性功能与尿流动力学,提高手术治疗效果。

基于TLR4/MyD88/NF-κB信号通路探讨左归丸、右归丸影响骨关节炎模型的作用机制研究

目的利用细胞、分子生物学技术,采用蛋白免疫印迹(Western blot,WB)、实时荧光定量反转录聚合酶链反应(Real-time PCR,RT-PCR)、酶联免疫吸附测定(Enzyme linked immunosorbent assay,Elisa)检测方法,观察左归丸与右归丸对大鼠膝骨关节炎(Knee osteoarthritis,KOA)软骨细胞、韧带组织的作用机制及细胞凋亡的影响,并讨论左归丸组与右归丸治疗膝骨关节炎的作用机制。方法选取8月龄的无特定病原体(Specific pathogen free,SPF)级别Wistar大鼠40只(250±20 g),雌鼠与雄鼠各20只,使用数字随机法分组,分为正常组、模型对照组、左归丸组、右归丸组、硫酸氨基葡萄糖组,每组各8只大鼠(雌雄各4只),造模方法为膝关节腔注射0.1 mL浓度为10%的木瓜蛋白酶溶液,每隔2 d造模1次,共进行3次造模,每次造模后驱赶大鼠30 min,用此方法对模型对照组、左归丸组、右归丸组、硫酸氨基葡萄糖组大鼠进行模型复制。模型复制成功后分别对5组大鼠进行相应干预。连续治疗4周后,所有大鼠取腹主动脉血后处死,Elisa法检测各组大鼠血清白细胞介素(IL)-1β表达水平,取出膝关节软骨及韧带组织,RT-PCR法与WB法检测软骨及韧带组织中toll样受体4(Toll-like receptor 4,TLR4)、IκB激酶(IKK)蛋白及mRNA的表达。剩余膝关节组织HE染色检测软骨组织情况。采用SPSS 22.0软件包对检测结果进行方差分析,P<0.05差异有统计学意义。结果(1)膝关节病理切片结果:经左、右归丸治疗后KOA炎症减轻。(2)RT-PCR检测结果:软骨和韧带组织中,较模型组对比左归丸组与右归丸组TLR4、IKK表达含量明显减少(P<0.05)。(3)WB检测结果:软骨和韧带组织中,较模型组对比左归丸组与右归丸组TLR4、IKK蛋白含量明显降低(P<0.05)。(4)Elisa法检测结果:各组大鼠血清中,左归丸组与右归丸组IL-1β表达水平明显降低(P<0.05)。结论左归丸与右归丸可通过抑制TLR4/髓样分化因子88(Myeloid differentiation factor 88,MyD88)/核因子κB(Nuclear factor-κB,NF-κB)信号通路活化,降低TLR4、IKK、IL-1β的表达,达到保护软骨细胞及韧带组织、控制炎症、促进受损组织恢复的目的。

督灸配合右归丸治疗肾阳虚型围绝经期综合征疗效观察

目的观察督灸配合右归丸治疗肾阳虚型围绝经期综合征的临床疗效及其对患者性激素、骨密度和生活质量的影响。方法将124例肾阳虚型围绝经期综合征患者随机分为治疗组和对照组,每组62例。对照组采用右归丸加味煎汤内服治疗,治疗组在对照组基础上采用督灸治疗。观察两组治疗前后各项中医证候积分、Kupperman绝经指数(Kupperman menopausal index,KMI)评分、围绝经期生存质量量表(menopause-specific quality of life,MENQOL)评分、血清性激素[促黄体生成素(luteining hormone,LH)、促卵泡激素(folliclestimulating hormone,FSH)、雌二醇(estradiol,E2)]和抗缪勒管激素(anti-Müllerian hormone,AMH)水平及骨密度变化情况,并比较两组临床疗效及不良反应发生率。结果治疗组总有效率为91.9%,明显高于对照组的74.2%,两组比较差异具有统计学意义(P<0.05)。两组治疗后各项中医证候积分、KMI评分、MENQOL评分及血清FSH、LH、AMH水平均较同组治疗前显著下降,血清E2水平及骨密度均显著上升,差异均具有统计学意义(P<0.05)。治疗组治疗后各项中医证候积分、KMI评分、MENQOL评分及各项性激素、AMH水平、骨密度与对照组比较,差异均具有统计学意义(P<0.05)。两组治疗期间不良反应发生率比较,差异无统计学意义(P>0.05)。结论督灸配合右归丸是一种治疗肾阳虚型围绝经期综合征的有效方法,能提高血清E2水平,抑制血清FSH、LH、AMH水平,增加骨密度,提高患者生活质量。

从命门水火学说探讨左归丸、右归丸对免疫系统的调节作用

“免疫”是机体抵抗异物,识别并处理自身异常细胞的一种防御机制,与中医学的“阳气”在功能上高度相似。且阳气主要发源于命门。张景岳基于其命门水火学说所创制的左归丸、右归丸为调补命门真阴、元阳之名方。因此,本文基于命门水火学说,结合现代研究进展,探讨左归丸、右归丸对免疫系统的调节作用。发现命门与中枢免疫器官在功能上存在高度相似性,可以说命门真阴是免疫活动物质基础的集中体现,骨髓中各类细胞分泌的细胞因子等生物活性分子是命门元阳的具体表现。左归丸、右归丸可以调补命门水火,使失衡的阴阳恢复到互根互用的平衡状态,对机体紊乱的免疫系统起到调节作用。使用左归丸、右归丸治疗一些以虚证为主证的自身免疫性疾病是切实可行的,值得进一步探究和推广。

右归丸联合中药熏蒸治疗肾阳虚型老年膝骨关节炎及其对关节液中OPN、S100A12及MMP-3的影响研究

目的探讨右归丸联合中药熏蒸治疗肾阳虚型老年膝骨关节炎及其对关节液中骨桥蛋白(OPN)、钙结合蛋白(S100A12)及基质金属蛋白酶-3(MMP-3)的影响。方法选用2018年1月—2020年5月老年肾阳虚型膝骨关节炎患者89例,将其作为观察对象。随机编号的方法分为对照组42例和联合组47例,全部完成治疗和随访。联合组选用右归丸联合中药熏蒸治疗;对照组选用常规疗法,持续治疗4周。比较两组患者的临床疗效,治疗前后分别记录骨关节炎调查量表(WOMAC)、视觉模拟量表评分(VAS)、生存质量测定量表(QOL-BREF)和关节液中OPN、S100A12、MMP-3的水平。结果与对照组比,联合组治疗2周、治疗4周WOMAC评分、VAS评分和QOL-BREF评分较低,差异有统计学意义(P<0.05);与对照组比较,联合组治疗2周、治疗4周关节液中OPN、S100A12和MMP-3水平较低,差异有统计学意义(P<0.05)。联合组的总有效率95.74%(45/42)高于对照组80.95%(34/42),差异有统计学意义(P<0.05)。联合组的恶心呕吐、腹泻、皮疹、肝肾功降低发生率与对照组比较,差异无统计学意义(P>0.05)。结论右归丸联合中药熏蒸治疗肾阳虚型老年膝骨关节炎的疗效确切,可以降低关节液中OPN、S100A12、MMP-3水平,从而提高生活质量,值得大力推广。

基于肝肾同源理论探讨右归丸治疗老年骨关节炎研究概况

骨关节炎是中国第二大致残性疾病,对中老年人生命安全及生活质量造成了极大的威胁,临床表现以关节疼痛、肿胀、畸形、活动障碍为主。近年来研究表明,临床上右归丸治疗肝肾亏虚型骨关节炎症效果显著,但其作用机制尚未完全明确。作者查阅近年来相关文献,将从“肝肾同源”理论入手对右归丸应用于骨关节炎的临床研究、基础研究及右归丸的药理作用机制进行探讨总结,旨在为“肝肾同源”理论指导下右归丸治疗骨关节炎提供治疗思路及参考。