Yinyanghuo(Herba Epimedii Brevicornus) and its components for chronic obstructive pulmonary disease:preclinical evidence and possible mechanisms

OBJECTIVE:To integrate Meta-analysis and bioinformatics strategies in the preliminary exploration of the potential mechanism of Yinyanghuo(Herba Epimedii Brevicornus) and its extract in treating chronic obstructive pulmonary disease(COPD).METHODS:First,Meta-analysis was carried out.The Chinese and English literature of Yinyanghuo(Herba Epimedii Brevicornus) in treating COPD was searched using the systematic strategy of combining subject words with free words.The included studies were evaluated by the SYRCLE risk bias assessment tool,after which the review manager software was used to combine the effect quantities for statistical analysis.Then,based on bioinformatics technology,the active ingredients and their targets of Yinyanghuo(Herba Epimedii Brevicornus) were screened,and the intersection genes were obtained by mapping and comparing with the targets of COPD.The "medicinal materials-compounds-targets model" was constructed,and the key pathways were annotated.Finally,the core target was docked with important compounds.RESULTS:A total of 8 studies were included in the Metaanalysis.The results showed that the Yinyanghuo(Herba Epimedii Brevicornus) group could significantly downregulate pro-inflammatory factors such as tumor necrosis factor-α(TNF-α) and interleukin(IL)-8 and increase the expression of anti-inflammatory factors and antioxidant factors such as IL-10 and phospho-protein kinase B(pAKT) in the COPD model(all P < 0.05).A total of 23 active components and 102 corresponding target genes of Yinyanghuo(Herba Epimedii Brevicornus) were obtained by bioinformatics technology,among which 17 compounds and 63 targets were closely related to COPD.The results of enrichment analysis mainly included TNF signaling pathway,phosphoinositide 3-kinase(PI3K)/Akt signaling pathway,cancer signaling pathway,and other inflammatory reactions,oxidative stress,and tumorrelated pathways.The molecular docking results showed that the binding energy fractions of the top five components of 24-epicampesterol with 10 core targets such as IL-6 were all less than ﹣5.0 kcal/mol,suggesting good binding ability.CONCLUSIONS:Meta-analysis and bioinformatics results indicated that the therapeutic effect of Yinyanghuo(Herba Epimedii Brevicornus) and its components on COPD might be related to antagonizing inflammation and oxidative stress.The above findings provide a preliminary basis for the development of Yinyanghuo(Herba Epimedii Brevicornus) as a natural drug for preventing and treating COPD.

Upregulation of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor axis by a combination of Yinyanghuo(Herba Epimedii Brevicornus) and Cheqianzi(Semen Plantaginis) improves erectile function in spontaneously hypertensive rats

OBJECTIVE:To evaluate the effects of a combination of Yinyanghuo(Herba Epimedii Brevicornus)(HEB)and Cheqianzi(Semen Plantaginis)(SP)on erectile dysfunction caused by essential hypertension in spontaneously hypertensive rats(SHRs),and to elucidate the role of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor(ACE2/Ang[1-7]/Mas receptor)axis in this process.METHODS:A total of 24 SHRs were randomly assigned to three groups:SHR-control,low-dose(12.5 g/kg)and high-dose(25 g/kg)HEB+SP(HEBSP).Eight Wistar-Kyoto rats were used as normal controls.HEBSP was administered by oral gavage for 28 d.Erectile function was measured once a week using the Heaton test.After 4 weeks of treatment,the corpus cavernosum was harvested from each rat to measure nitric oxide(NO),nitric oxide synthase(e NOS)and Ang(1-7)levels,as well as ACE2,Mas receptor and neuronal nitric oxide synthase(n NOS)protein expression.RESULTS:After 4 weeks of treatment,HEBSP significantly increased erectile function in the treated group compared with SHR-control group(P<0.01).Additionally,HEBSP treatment significantly increased cavernosal levels of Ang(1-7),e NOS and NO.Moreover,HEBSP significantly elevated the expression levels of ACE2,Mas receptor and n NOS.These beneficial effects were elevated in the high-dose HEBSP group.CONCLUSION:HEBSP improved erectile function in SHRs by upregulating the ACE2/Ang(1-7)/Mas receptor axis,e NOS and n NOS pathways.

Efficacy of an Yinyanghuo(Herba Epimedii Brevicornus)-Xianmao(Rhizoma Curculiginis)drug pair in a rat model of polycystic ovary syndrome

OBJECTIVE:To identify active compounds in an Yinyanghuo(Herba Epimedii Brevicornus)-Xianmao(Rhizoma Curculiginis)drug pair(ECD)and investigate its efficacy on polycystic ovary syndrome(PCOS),and its possible mechanism in a rat model of PCOS.METHODS:A network pharmacology approach involving a characteristic drug assessment,active compound and target prediction,PCOS gene collection as well as network analysis was employed.The ovary morphology after treatment was observed using an animal model and western blotting and real-time PCR were used to verify AKT1 as the molecular target.RESULTS:Six networks were constructed,an active compound-target network for the ECD(C-T network),a drug-target network(D-T network),a related genes network,a targets interaction network,a key genes interaction network,and a gene-pathway network.A total of 41 compounds and 261 targets were identified for the ECD,232 PCOS-related genes,31 cogenes,and 14 pathways.These pathways may be involved in the efficacy of ECD on PCOS.The proteins most involved in the signal pathways for all targets were AKT1,IL6,INSR,ESR,and GSK3B.The AKT1 target was selected for experimental verification.Based on the Western blot and real-time PCR results,the expression of AKT1 in the PCOS model varied after treatment with ECD.CONCLUSIONS:Our findings suggest that the ECD can reverse the negative morphological changes in ovarian tissue that occur in model rats of PCOS.AKT1 may be a key mediator of the observed ability of the ECD to protect against PCOS in the model rats.

Effects of Herba Epimedii and Fructus Ligustri lucidi on the Transcription Factors in Hypothalamus of Aged Rats

比较地用蛋白质在视下丘在 54 个抄写因素的表示分析差别的目的削为沈跟随中国药的药()-tonification, 草 Epimedii，和到年老的老鼠的 Fructus Ligustri lucidi (FL ) 。

Protective effects of kidney-tonifying Chinese herbal preparation on substantia nigra neurons in a mouse model of Parkinson's disease

The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson's disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson's disease mouse models. Rhizoma Polygonati could downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustri Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustri Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson's disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.

淫羊藿与女贞子配伍对自噬激活状态ASMCs的影响

目的研究淫羊藿、女贞子配伍对自噬激活剂雷帕霉素(rapamycin,RAP)诱导的气道平滑肌细胞(airway smooth muscle cells,ASMCs)自噬、凋亡及增殖活性的调节作用。方法培养大鼠原代ASMCs,随机分为正常组、RAP组、淫羊藿组、女贞子组、配伍组。根据分组分别加入正常大鼠血清或含药大鼠血清;除正常组外,其余各组同时加入RAP诱导的ASMCs,48h后进行检测。电镜观察细胞超微结构;MTT法检测细胞活力;免疫细胞化学法检测增殖蛋白ki-67蛋白表达,评价细胞增殖活性;Annexin V-FITC/PI法检测细胞凋亡活性;Western blot法和细胞免疫荧光法检测LC3、Beclin-1、mTOR、p-mTOR、Bax、Bcl-2、caspase-3及p53蛋白表达。结果淫羊藿、女贞子配伍能显著抑制RAP诱导的ASMCs自噬,下调LC3-Ⅱ/Ⅰ比值(P<0.05)及Beclin-1蛋白表达(P<0.01);改善RAP诱导的细胞活力、增殖活性降低(P<0.01);抑制凋亡水平升高(P<0.01),且优于淫羊藿、女贞子单用组(P<0.05或P<0.01)。此外,配伍组能上调caspase-3蛋白表达(P<0.05),下调p53蛋白表达(P<0.01);但对mTOR及p-mTOR、Bax及Bcl-2蛋白表达无显著影响。结论淫羊藿、女贞子配伍能抑制RAP诱导的ASMCs的过度自噬/凋亡水平,恢复ASMCs的细胞活力及增殖活性,避免ASMCs过度损伤。

谭新华教授治疗勃起功能障碍用药规律分析

目的通过对谭新华教授治疗勃起功能障碍的中药处方进行用药和配伍规律分析,为临床治疗提供参考。方法收集谭新华专家门诊2021年1月至2021年10月治疗勃起功能障碍的中药处方,建立医案数据库,基于中医传承计算平台3.0进行药物性味、归经、使用频次分析,以及相关规则与聚类分析。结果纳入治疗勃起功能障碍的中药处方227张,包含药物153味。用药以甘温为主,归肝、肾两经居多,脾经其次。补虚药使用最多,频次≥100的药物有10味,高频药物有茯苓、巴戟天、淫羊藿等,常见药对是淫羊藿-巴戟天。聚类分析发现,谭新华教授治疗勃起功能障碍有脾肾同调、补肾固涩、疏肝活血、平补阴阳、肝肾并治5种经典治法。结论谭新华教授治疗勃起功能障碍宜清补脾胃,在补肾的基础上注重固肾、疏肝养血、气火同调,以期为临床治疗用药提供思路和参考。

淫羊藿总黄酮对大鼠实验性骨质疏松生化学指标的影响

目的 探讨淫羊藿总黄酮防治大鼠实验性骨质疏松症过程中骨代谢生化指标的变化及其意义。方法 采用维甲酸灌胃造成大鼠实验性骨质疏松症模型 ,检测灌服高、中、低 3种剂量的淫羊藿总黄酮后大鼠血清生化指标、尿生化指标、骨矿含量及骨密度指标的变化 ,并与模型组、正常对照组和阳性对照组进行比较。结果 3个剂量组的血清E2 、T和BGP水平均高于模型组 ,尿Ca/Cr、DPD和血清PTH水平均低于模型组 ,股骨Ca、P和骨密度与正常对照组接近 ,与模型组差异有显著性。结论 淫羊藿总黄酮对维甲酸造成的大鼠骨质疏松症有明显的防治作用 ,骨代谢生化指标变化情况与骨密度的变化相一致 。

择时服用淫羊藿对性激素水平的影响

为了证实《黄帝内经》“择时服药”理论的科学性,本研究通过择时服用淫羊藿对幼鼠性激素水平的影响,探讨服药时间与药效的关系。结果显示:(1)淫羊藿煎剂具有性激素样作用,可以使雌性小鼠子宫增重,雌二醇(E_2)含量升高,雄性小鼠血清睾酮(T)含量升高。(2)卯时、酉时服药对雌性小鼠子宫增重,E_2含量升高,雄性小鼠血清T含量升高有不同程度影响,卯时作用强于酉时。表明补肾壮阳药清晨服用效果最佳。

淫羊藿研究进展

对国内外药用淫羊藿的化学成分、质量控制、炮制工艺、提取工艺及其对免疫系统、心脑血管系统、生殖系统等的作用进行了综述。

淫羊藿总黄酮抗大鼠实验性骨质疏松作用研究

目的 研究淫羊藿总黄酮对维甲酸所致大鼠骨质疏松的防治作用并探讨其主要作用机理。方法 用70mg·kg-1·d-1维甲酸连续灌胃 14d ,诱发大鼠骨质疏松模型 ,用高、中、低 3种剂量的淫羊藿总黄酮进行治疗 ,观测大鼠器官指数、骨计量学指标及骨组织形态计量学的变化。结果 淫羊藿总黄酮各剂量组前列腺和睾丸指数均明显增大并恢复到正常对照组水平 ,脾脏和肾上腺的代偿性肥大消失 ,股骨的干重、灰重、骨钙、骨磷和骨密度等均明显高于模型组 ,胫骨近端的骨小梁面积百分比、骨小梁厚度、骨小梁间隙以及胫骨中段的皮质骨面积百分比等亦较模型组有显著升高 ,第一腰椎中骨小梁指数的变化与胫骨松质骨近似。结论 淫羊藿总黄酮可通过保护性腺、抑制骨吸收和促进骨形成等途径 ,使机体骨代谢处于骨形成大于骨吸收的正平衡状态 ,抑制骨量丢失 。

淫羊藿总黄酮的提取分离工艺研究

目的 考察淫羊藿总黄酮的最佳提取、分离、纯化工艺。方法 采用四因素三水平正交实验进行提取条件的优选 ;用乙酸乙酯萃取、酸碱沉淀和铅盐沉淀三种分离方法对分离工艺进行比较 ;分别用 95 %乙醇、85 %乙醇、正丁醇、乙酸乙酯和甲醇对提取物进行纯化。结果 影响淫羊藿总黄酮提取率的因素依次为溶媒、提取次数、提取时间、提取溶剂的用量。最佳提取工艺为 10倍量 70 %乙醇提取 3次 ,每次 1 5h ;分离工艺考察表明 3种分离方法中以酸碱沉淀法较为理想 ;提取物经甲醇纯化后总黄酮含量和收率均明显提高 ,甲醇可作为理想的纯化溶剂。结论 该实验确定的最佳提取工艺稳定性好且简便易行 ,适合工业化大生产。

氟苯尼考和中草药黄芪淫羊藿合剂对鸡体液免疫反应的影响

试验分别对免疫期间的各组鸡连续两周给予不同剂量的氟苯尼考、黄芪淫羊藿合剂后,在不同日龄采取血样,用血凝抑制(HI)试验分别对鸡新城疫(ND)抗体水平和禽流感(AI)抗体水平进行监测。结果表明,混饮给药,小剂量(50mg/L)氟苯尼考对ND抗体水平影响不显著(P>0 05),而中(150mg/L)、高(250mg/L)剂量氟苯尼考对ND抗体产生有显著的抑制作用(P<0 01);氟苯尼考追加给药1周后,中、高剂量组ND抗体水平又有显著降低(P<0 05),而小剂量组ND抗体水平差异不显著;中(15ml/L)、高(22 5ml/L)剂量黄芪淫羊藿合剂对ND、AI抗体产生有显著增强作用(P<0 01);对因氟苯尼考造成的免疫抑制,黄芪淫羊藿合剂可使抗体水平恢复至对照组水平(P>0 05)。

淫羊藿对抗肾上腺皮质激素不良反应的实验研究

淫羊藿水提液对长期应用肾上腺皮质激素所致的“肾阳虚”等不良反应,特別是肾上腺萎缩与骨质疏松症,有显著的防治作用。

高效液相色谱法测定淫羊藿中淫羊藿定C和淫羊藿苷的含量

目的 :测定不同种和不同产地的淫羊藿中淫羊藿定C和淫羊藿苷的含量 ,探讨淫羊藿药材的质量控制方法。方法 :色谱柱为HypersilBDS C18(4mm× 2 50mm ,5μm ) ,以乙腈 0 .0 5%磷酸作为梯度洗脱流动相 ,G 13 15A光电二极管阵列检测器 ,检测波长 2 72nm。结果 :淫羊藿定C和淫羊藿苷平均回收率分别为 99.7% ,10 2 .5% ,RSD分别为 1.5% ,1.1%。结论 :本方法快速 ,重现性和稳定性好 。

淫羊藿总黄酮对成骨细胞中OPG和RANKL mRNA基因表达影响的实验研究

目的:探讨淫羊藿总黄酮对大鼠成骨细胞中骨保护素(OPG)和核因子κB受体激活因子配体(RANKL)mRNA基因表达的影响。方法:体外培养成骨细胞,分别加入含0·1、1、10、100ng/ml淫羊藿总黄酮(HEF)的培养液,48h及96h后分别提取细胞总mRNA,用RT-PCR方法分析OPG/RANKL的mRNA的表达,进行半定量分析。结果:培养48h后,与对照组比较,HEF增强了OPGmRNA的表达,有显著性差异(P<0·05或P<0·01),VOPG/VRANKL比值增大;96h后,HEF明显增强了OPGmRNA的表达,各浓度组与对照组相比,均有显著性差异(P<0·05或P<0·01),各浓度组VOPG/VRANKL比值增大,有显著性差异(P<0·05或P<0·01)。结论:淫羊藿总黄酮可能通过增加成骨细胞OPG的表达来抑制破骨细胞的分化和成熟,从而抑制骨吸收,达到治疗骨质疏松症的目的。

紫外分光光度法和高效液相色谱法测定淫羊藿总黄酮含量的比较研究

目的:比较紫外分光光度法和高效液相色谱法测定淫羊藿总黄酮含量的差异,探讨淫羊藿总黄酮含量测定方法的可靠性。方法:紫外分光光度法以淫羊藿苷为指标性成分,在270 nm 波长处进行测定。高效液相色谱法以 ZORBAX SB-C_(18)柱(250mm×4.6 mm,5μm)为分析柱,乙腈-水梯度洗脱,流速1.0 mL·min^(-1),检测波长为270 nm。经紫外光谱识别的黄酮类成分(部分经 ESI-MS 确认),除朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ和宝藿苷Ⅰ以自身对照外,其他黄酮类成分均以淫羊藿苷为参比进行定量,淫羊藿总黄酮含量等于朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ、宝藿苷Ⅰ和其他黄酮成分含量之和。结果:紫外分光光度法,淫羊藿苷在2.45～24.50μg·mL^(-1)范围内线性关系良好(r=0.9996),测得淫羊藿总黄酮含量以淫羊藿苷计为56.6%。高效液相色谱法中,与淫羊藿苷紫外光谱相似的33个色谱峰,MS 归属了其中10个主要成分均为黄酮类成分。朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ和宝藿苷Ⅰ分别在0.093～1.852μg(r=0.9998)、0.107～2.136μg(r=0.9997)、0.094～1.876μg(r=0.9998)、0.098～1.956μg(r=0.9998)线性关系良好,淫羊藿总黄酮含量为35.6%。结论:紫外分光光度法和高效液相色谱法测定的淫羊藿总黄酮含量差异显著,紫外分光光度法的准确性有待进一步考证。

淫羊藿总黄酮对大鼠成骨细胞代谢调控的机制研究

目的:探讨淫羊藿总黄酮对大鼠成骨细胞代谢调控的影响。方法:取新生SD大鼠的颅骨,胶原酶法培养成骨细胞。应用MTT法、对硝基苯磷酸盐法(PNPP)和RT-PCR方法观察淫羊藿总黄酮对体外培养成骨细胞的增殖、碱性磷酸酶活性和OPG mRNA,RANKL mRNA表达的影响。结果:淫羊藿总黄酮能刺激成骨细胞增殖,提高碱性磷酸酶活性,增强OPG mRNA的表达(P<0.05或P<0.01)。结论:淫羊藿总黄酮能促进成骨细胞的增殖、分化,增加OPG mRNA的表达,对RANKL的表达无明显影响。

HPLC法同时测定淫羊藿中朝藿定A、B、C与淫羊藿苷的含量

建立了同时测定淫羊藿中朝藿定A、B、C与淫羊藿苷含量的高效液相色谱方法.淫羊藿样品经超声波提取,以50%（ψ）乙醇溶液为溶剂,在温度50℃、料液比1∶60条件下超声波提取30 min,共提取2次.色谱条件：ZORBAX SB-C8色谱柱（5 μm,4.6×250 mm）;流动相A为乙腈,B为1%乙酸溶液;梯度洗脱;紫外检测波长为270 nm.上述4种类黄酮的标准曲线在6.6～33.0 mg/L（朝藿定A、淫羊藿苷）或6.6～55.0mg/L（朝藿定B、C）范围内呈良好的线性关系（r＞0.99）;加标回收率在97%～102%之间;相对标准偏差小于2%（n=5）.测得不同产地心叶淫羊藿叶片中4种类黄酮的质量分数差异较大;朝藿定A广泛分布于淫羊藿属中.本方法在淫羊藿药材质量控制中具有一定的参考价值.

淫羊藿对大鼠下丘脑—垂体—肾上腺—胸腺轴抑制模型的影响

本研究用皮质酮造成大鼠下丘脑-垂体-肾上腺-胸腺(HPAT)轴抑制模型,观察补肾助阳要药淫羊藿对受抑大鼠的调节作用。结果表明,模型大鼠垂体、肾上腺、胸腺重量明显减轻;下丘脑室旁核小细胞区促肾上腺皮质激素释放因子(CRF)阳性细胞及正中隆起CRF阳性神经纤维及垂体前叶促肾上腺皮质激素(ACTH)阳性细胞明显减少,胞体变小,染色变淡,肾上腺及胸腺小体明显减少;血浆ACTH、皮质酮以及淋巴细胞增殖反应、自然杀伤细胞活性(NKCC)、γ-干扰素(γ-IFN)诱生能力较正常组明显降低。淫羊藿灌胃能有效改善皮质酮对HPAT轴形态与功能的抑制。提示淫羊藿对神经内分泌免疫的调节起了重要作用。