LC-MS determination and pharmacokinetic study of salidroside in rat plasma after oral administration of suspensions of traditional Chinese medicine Erzhi Wan and Fructus Ligustri lucidi

A simple,rapid and sensitive liquid chromatography-mass spectrometry（LC-MS）method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic ether-isopropanol（2∶1）and the extraction was performed on a Kromasil C18 column（150 mm×4.6 mm,5 μm）with the mobile phase of methanol-water（41∶59,v/v）within a run time of 6.0 min.The analyte was monitored with positive electrospray ionization（ESI）by selected ion monitoring（SIM）mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard（IS）geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.

Application Progress of Fructus Ligustri Lucidi( FLL) in Animal Husbandry

Fructus Ligustri Lucidi （ FLL）, a well-known and commonly used Chinese herbal medicine in clinical practice, is often applied in animal husbandry in the form of monomer or compound. The chemical composition, pharmacological effects of FLL and its application progress in animal husbandry are reviewed.

Compatibility with Fructus Ligustri Lucidi Effectively Mitigates Idiosyncratic Liver Injury of Epimedii Folium by Modulating NOD-like Receptor Family Pyrin Domain Containing 3 Inflammasome Activation

Background: Idiosyncratic drug-induced liver injury(IDILI) is a serious side effect of drugs, Epimedii Folium(EF) is unequivocally implicated in idiosyncratic liver injury onset, potentially due to its ability to perturb the NOD-like receptor family pyrin domain containing 3(NLRP3) inflammasome. Fructus Ligustri Lucidi(FLL), a frequently used medicinal combination with EF, has not yet been investigated for its ability to ameliorate EF-associated hepatotoxicity. Aims and Objectives: Study on the mechanism of compatibility of FLL to alleviate liver injury caused by EF. Materials and Methods: Western blot was used to determine the expression of related proteins, ELISA was used to detect the secretion of related inflammatory factors IL-1β, IL-18, IL-6 and TNF-α, liver injury indexes were detected and liver pathological tissue staining was used to evaluate the liver injury. Results: Our results demonstrated that EF exerted a particular augmenting effect on the stimulation of the NLRP3 inflammasome mediated by nigericin or ATP, whereas FLL suppressed the NLRP3 inflammasome stimulation. Furthermore, an equal EF to FLL ratio significantly reduced the stimulatory effects of EF. Moreover, EF has the potential to induce hepatic injury and augment pro-inflammatory cytokine synthesis in rats subjected to LPS. However, when combined with FLL, the detrimental effects of EF were mitigated. Conclusions: FLL possesses the capacity to attenuate EF-associated hepatotoxicity by suppressing EF-triggered NLRP3 inflammasome activation. Thus, FLL holds promise for improving the clinical safety profile of EF, shedding light on the potential of compatibility and detoxification theories in traditional Chinese medicine.

女贞子中主要活性成分红景天苷和特女贞苷的逆转铁过载促进骨形成相互作用研究

目的评价女贞子中主要活性成分红景天苷和特女贞苷的逆转铁过载促进骨形成抗绝经后骨质疏松(postmenopausal osteoporosis,PMOP)效应,并探讨其相互作用关系。方法建立柠檬酸铁铵诱导的斑马鱼骨质疏松模型,分别评估红景天苷(1、5、25、50、100μmol/L)、特女贞苷(1、5、25、50、100μmol/L)及其两种成分混合溶液(1∶1)对骨矿化的影响;茜素红染色后以骨骼染色面积和累积光密度评价其骨矿化程度,并运用等效线法和等辐射分析法评价红景天苷、特女贞苷抗PMOP的相互作用。结果红景天苷、特女贞苷及其混合溶液均有较好地逆转铁过载所致的骨形成抑制作用,且在1~100μmol/L内呈现显著的浓度依赖性;在等效线法中,经半数有效浓度(EC50)计算得混合组γ>1,表明二者无明显的协同作用;在等辐射分析法中,混合组中相应成分的EC50比值分数总和为1.16,其复合点落在95%可信限内(P>0.05),表明两者呈相加作用关系,且未见明显协同作用。结论红景天苷和特女贞苷均可逆转铁过载诱导的骨形成抑制,且当红景天苷和特女贞苷联合使用时,两者呈相加作用。

女贞子活性成分防治骨质疏松症的分子机制

背景:目前,中医药已被证实在抗骨质疏松方面具有显著作用,女贞子活性成分的抗骨质疏松有效性及作用机制逐渐得到学者们的关注。目的:分析总结女贞子活性成分在体内外发挥抗骨质疏松方面的研究进展。方法:检索中国知网及PubMed数据库收录的相关文献,中文检索词为“骨质疏松症,骨髓间充质干细胞,成骨细胞,破骨细胞,女贞子,信号通路”;英文检索词为“Osteoporosis,Bone marrow mesenchymal stem cells,Osteoblast,Osteoclast,Ligustri Lucidi Fructus,Signal path”,并根据研究需要确立相应的标准,对所得文献进行筛选,最终纳入82篇文献进行综述。结果与结论:女贞子活性成分在体内外发挥抗骨质疏松作用主要涉及以下几种重要成分:①红景天苷通过抑制骨硬化蛋白(SOST)和Dickkopf相关蛋白1(Dkk1)的表达,激活了Wnt/β-catenin信号通路,该激活过程增强了去卵巢大鼠和原代成骨细胞中磷酸化低密度脂蛋白受体相关蛋白6(p-LRP6)的表达,同时降低了糖原合酶激酶3β的表达。进一步地,红景天苷促进了细胞核内β-catenin、特异性转录因子2(Runx2)和骨髓细胞瘤病毒癌基因(c-Myc)的表达,从而促进了成骨细胞的骨形成能力。红景天苷可直接作用于成骨细胞,促进骨形成,为骨质疏松症的治疗提供了新的策略。②橄榄苦苷显著提高了SD大鼠的骨密度,并通过降低终末白细胞介素6和碱性磷酸酶浓度来调节骨代谢。体外实验表明,橄榄苦苷促进了成骨细胞的增殖,并上调了骨保护素蛋白和mRNA的表达,同时抑制了核转录因子κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)蛋白和mRNA的表达。这一作用机制与调节骨保护素/RANKL系统的平衡密切相关,展现出能提高骨密度,又能通过调节骨代谢相关因子来维护骨骼健康的优势,但对Ca2+浓度无显著影响,可能限制了其在某些特定骨质疏松类型中的应用。③松果菊苷通过降低磷脂酰肌醇3-激酶(phosphatidylinositol 3 kinase,PI3K)的表达、减少蛋白激酶B(protein kinase B,Akt)的磷酸化程度和破骨特殊标记蛋白(c-Fos)的表达,有效抑制了破骨细胞中PI3K/Akt/c-Fos通路的激活。这一抑制作用降低了破骨细胞的增殖和成熟能力,从而有助于减少骨吸收,其优势能直接针对破骨细胞进行干预,为骨质疏松症的治疗提供了新的靶点,但是目前对破骨细胞的具体调控机制尚需深入研究,且其长期效果和安全性需进一步评估。④女贞子活性成分在骨质疏松方面展现出良好的治疗效果,但其作用机制复杂,涉及多个基因、蛋白和信号通路的相互作用,未来需开展大规模的临床试验以验证其有效性和安全性,同时需要进一步探索女贞子活性成分与其他药物的联合应用策略,以期获得更好的治疗效果。

黑豆与黄酒共制女贞子炮制工艺优化

目的优化黑豆与黄酒共制女贞子炮制工艺。方法在单因素试验基础上,以蒸制时间、蒸制次数、烘制温度为影响因素,外观性状,红景天苷、特女贞苷、女贞苷G13、醇溶性浸出物含量,DPPH、ABTS自由基清除率的总评“归一值”(OD值)为评价指标,星点设计-响应面法优化炮制工艺。结果最优工艺为蒸制时间70 min,蒸制次数7次,烘制温度90℃,OD值为0.8415。结论该方法简便可靠,稳定可行,可用于黑豆黄酒共制女贞子。

1994-2023年女贞子研究文献可视化分析

目的通过文献计量学方法分析女贞子研究现况及趋势,为相关研究提供参考。方法检索中国知识资源总库(CNKI)、中文科技期刊数据库(VIP)、中国学术期刊数据库(万方数据)、中国生物医学文献数据库(CBM)、Web of Science、PubMed 1994年1月1日-2023年8月13日收录的女贞子研究文献。采用NoteExpress3.6.0.9155软件管理并筛选文献,分析发文时间趋势,采用CiteSpace6.2.R4和VOSviewer1.6.18.0软件分析纳入文献作者、来源期刊、研究机构和关键词,并绘制知识图谱。结果纳入795篇中文文献和175篇英文文献,年发文量均有所波动。中、英文文献发文较多的机构分别为首都医科大学和香港理工大学,发文较多的作者分别为刘仁慧和Wong Mansau,发文较多的期刊分别为《中国实验方剂学杂志》、Phytotherapy Research和Journal of Ethnopharmacology。高频关键词包括齐墩果酸、特女贞苷、炮制、抗氧化、提取工艺和大鼠等;生成24个聚类与44个突现关键词。结论该领域研究热点包括齐墩果酸和特女贞苷等药理成分的提取、纯化,女贞子和淫羊藿、虎杖等中药配伍,延缓衰老、治疗癌症等疾病的疗效与机制,栽培与质量控制等。

女贞子-墨旱莲药对提取物的指纹图谱和化学成分鉴定研究

目的建立女贞子-墨旱莲药对提取物的HPLC指纹图谱以控制其质量,并对提取物中所含有的化学成分进行结构鉴定。方法采用HPLC法,固定相为ZORBAX Extend C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-体积分数为0.1%甲酸溶液梯度洗脱,流速为1.0 mL·min^(-1),检测波长为265 nm,柱温为35℃,进样量为10μL;建立15批不同产地女贞子-墨旱莲药对提取物的HPLC指纹图谱,对其进行相似度评价、聚类分析和正交偏最小二乘方判别分析;采用超高效液相色谱-静电场轨道阱高分辨质谱(UPLC-Q-Exactive Orbitrap MS)联用技术,分别在正、负离子扫描模式下对提取中的化学成分进行检测,基于高分辨质谱一级和二级数据进行结构鉴定。结果由不同产地药材随机组合的15批女贞子-墨旱莲药对提取物的相似度介于0.865~0.992之间,药对指纹图谱中共确定了16个共有峰,指认了其中7个色谱峰;液质联用共检测到55个化合物,采用对照品准确鉴定了7个成分的化学结构,并推测了其它48个化合物的可能化学结构。结论建立的指纹图谱方法简便可靠,重复性好,可用于女贞子-墨旱莲药对提取物的质量控制研究;UPLC-Q-Exactive Orbitrap MS联用技术可实现对女贞子-墨旱莲药对提取物中化合物的结构鉴定,为其后续研究奠定了基础。

女贞子活性成分代谢特征及药理作用研究进展

女贞子包含五环三萜类、苯乙醇苷类、环烯醚萜类及黄酮类等多种成分,同时还含挥发油、多糖和脂肪酸等成分,其具备多种药理活性表现,例如抵抗癌症、预防骨质疏松、减缓炎症止痛、保护肝脏、促进头发生长和明目等,但因其口服生物利用度低,有关其在体内外的吸收代谢过程及其药理作用机制研究较少,极大地影响了女贞子的后续研究。因此,对女贞子中主要的活性成分、体内外吸收代谢及其药理作用进行综述;女贞子主要化学成分多于肝脏和肠道代谢,体内吸收方式有主动运输和被动扩散等,代谢方式主要为氧化和葡萄糖醛酸化等,旨在为深入开发女贞子药用价值提供参考。

基于CiteSpace和VOSviewer对女贞子的研究现状与热点分析

利用CiteSpace和VOSviewer对女贞子近三十年的中文文献进行计量学分析,以探究女贞子的研究现状及发展趋势。本研究整理收集中国知网、维普、万方及中国生物医学文献数据库中近三十年(1993年至2023年)女贞子相关研究文献,通过Java语言环境下的VOSviewer与CiteSpace软件对其发文量、作者、发文机构、关键词进行可视化分析。本文纳入的1 240篇文献可视化,结果显示,发文量最多的作者是刘慧仁(首都医科大学);发文量最多的机构是山东中医药大学(22篇);通过对关键词的分析发现,女贞子的研究热点集中在化学成分的测定、质量标准的制定、药理作用的探索、配伍机制的研究、“女贞子-墨旱莲”(二至丸)的配伍、临床经验的总结等方面。当前,女贞子研究的重点和热点是复方(中药)/治疗应用、化学成分分析和“二至丸”配伍分析。

基于中医“肾-脑-骨”轴探讨女贞子及其活性成分的多脏器药理作用及潜在调控机制

女贞子作为一种传统的补益肝肾中药,在中医药临床应用中具有重要地位。随着现代科技的发展,对女贞子的药理学活性研究也日益深入。“肾-脑-骨”轴的概念源于“肾主骨,生髓,通于脑”的中医理论,女贞子的多脏器药理学活性与该理论内涵具有紧密的契合点。基于“肾-脑-骨”轴的理论基础,旨在探讨女贞子及其活性成分的多脏器药理学作用及潜在调控机制,以期为临床应用与药物开发提供理论依据。

Synergistic effects of combined therapy of curcumin and Fructus Ligustri Lucidi for treatment of osteoporosis: cellular and molecular evidence of enhanced bone formation

Objective: The present study explored the effects of the combined herbal therapy consisting of curcumin(CUR) and Fructus Ligustri Lucidi(FLL) on aspects of bone regeneration.Methods: Prior to analyzing the ability of this novel combined herbal therapy to promote aspects of bone regeneration, its cytotoxicity was determined using MC3 T3-E1 cells(pre-osteoblast model). Cell proliferation was evaluated using phase-contrast microscopy and cell differentiation was estimated using alkaline phosphatase activity. The effect of the combined herbal therapy(CUR + FLL) was also assessed in terms of mineralization in the extracellular matrix(ECM) of cultured cells. Further, to explore the molecular mechanisms of bone formation, time-dependent expression of bone-regulating protein biomarkers was also evaluated.Results: Combined herbal therapy(CUR + FLL) significantly upregulated the viability, proliferation and differentiation of MC3 T3-E1 cells compared to the monotherapy of CUR or FLL. The magnitude of ECM mineralization(calcium deposition) was also higher in MC3 T3-E1 cells treated with combined therapy.The time-dependent expression of bone-forming protein biomarkers revealed that the tendency of expression of these bone-regulating proteins was remarkably higher in cells treated with combined therapy.Conclusion: The co-administration of CUR and FLL had superior promotion of elements of bone regeneration in cultured cells, thus could be a promising alternative herbal therapy for the management of bone erosive disorders such as osteoporosis.

女贞子防治骨质疏松症药理机制的研究

骨质疏松症是常见的骨代谢疾病,随着中国人口老龄化程度的不断加重,已成为广泛关注的公共健康问题。女贞子作为传统的补肾中药,对骨质疏松症的治疗有着独特优势。研究显示女贞子具有骨保护作用,能够减少骨质流失,改善骨质疏松症,但其作用机制尚不清晰。为此,本文利用中国知网、PubMed等数据库,以骨质疏松、女贞子、成骨等为关键词,检索了近15年国内外相关文献。根据现有研究,本文主要从钙平衡、氧化应激、骨代谢及激素样作用4个方面综合论述女贞子防治骨质疏松症的机制,以期为女贞子抗骨质疏松的进一步研究提供参考及为中药防治骨质疏松症提供思路。

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

Objective： To comparatively analyze the difference in the expression of 54 transcription factors in the hypothalamus using protein chips following the medication of Chinese drugs for Shen （肾）-tonification, Herba Epimedii, and Fructus Ligustri lucidi （FL） to aged rats. Methods： Wistar rats, aged 15 months of SPF grade, were randomized into three groups, three males and three females in each group. They were medicated with Herba Epimedii decoction （HED, 0.14 g/kg）, Fructus Ligustri lucidi decoction （FLD, 0.12 g/kg）, and distilled water, respectively, twice a day for 15 days. The rats were sacrificed at the morning of the 16th day 1 h after medication, and their hypothalamus was taken and made into homogenate under an ice-bath for detecting the expression of transcription factors with chip technique. Results： The expressions of signal transduction and transcription activation factor-6 （Stat-6） and androgen receptor （AR） were up-regulated, and those of pre-B transcription factor1 （Pbx-1）, stat-1 and AP-2 were down-regulated in both HED and FLD treated groups, but these changes occurred mainly in female rats in the former while mainly in males in the latter. Conclusions： Chinese drugs for Shen-tonification could impact the expression of transcription factors in the hypothalamus of aged rats, dominantly on the neuro-endocrine factors responsible for the growth and development. The effects of drugs for tonifying Shen-yang and for tonifying Shen-yin are different, which is probably one of the pharmacological mechanisms of the Shen-tonifying drugs.

不同种源女贞子药用有效成分比较

为探讨女贞子红景天苷、特女贞苷等药用有效成分在种源水平上的遗传变异规律及地理变异规律,以湖南芷江、湖南新宁、四川峨眉山、云南昭阳、云南晋宁等5个种源为材料,比较了不同种源女贞子药用有效成分含量的差异。结果表明:女贞子红景天苷、特女贞苷、木犀草苷、齐墩果酸等4种药用有效成分含量在种源间均存在极显著差异;女贞子红景天苷、特女贞苷、木犀草苷、齐墩果酸的含量在种源水平上能稳定遗传,且受外界环境条件的影响比较小;4种药用有效成分含量之间的相关性均未达到显著水平;以红景天苷、特女贞苷、木犀草苷和齐墩果酸含量为指标,不同种源女贞子药用有效成分含量的综合隶属度排序依次为湖南芷江、云南晋宁、云南昭阳、湖南新宁、四川峨眉山,湖南芷江种源在药用有效成分含量上具有优良表现。

Anti-diabetic Activity of Zhenqing Recipe and Ligustri Lucidi Fructus in Type 2 Diabetic Rats

Objective To investigate the influence of Zhenqing Recipe(ZQR)and Ligustri Lucidi Fructus(LLF)on diabetic rats and its possible mechanism.Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats.The model rats were randomly divided into three groups:diabetic model,ZQR-treated,and LLF-treated groups for 8-weeks treatment.The normal Wistar rats were as a normal control group.Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group(P<0.01,0.05,respectively).Both ZQR and LLF markedly reduced serum triglycerides(P<0.01,0.05,respectively),and increased the insulin sensitivity index(P<0.05).Histopathology revealed that ZQR and LLF reduced pancreatic damage.Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group(P<0.01,0.05,respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group(P<0.01).Conclusion ZQR has therapeutic effect on type 2 diabetes,it ameliorates the histopathological changes of pancreas,protectsβcells,improves insulin resistance,and attenuates the expression of SREBP-1c.This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.

基于网络药理学探讨女贞子治疗糖尿病肾病作用机制

目的 探究女贞子治疗糖尿病肾病(DN)的作用机制。方法 通过网络药理学数据库筛选女贞子和DN作用靶点,进行蛋白互作(PPI)、基因本体论(GO)数据库和京都基因与基因组百科全书(KEGG)数据库分析。结果 得到女贞子作用于DN的关键靶点18个,分别与细胞转录、生长、信号转导、炎症等相关;KEGG富集分析发现,女贞子与晚期糖基化产物、动脉粥样硬化、炎症因子等通路密切相关。结论 初步探究了女贞子治疗DN的作用机制,为后续实验提供理论基础。

贞芪扶正胶囊及其中药配方颗粒、传统饮片有效成分比较研究

目的比较贞芪扶正胶囊(女贞子、黄芪)及其中药配方颗粒、传统饮片的有效成分差异。方法采用高效液相色谱法(HPLC)定量分析,比较贞芪扶正胶囊及其对应比例重量的饮片和配方颗粒中主要有效成分(毛蕊异黄酮葡萄糖苷、特女贞苷、黄芪甲苷)含量差异,根据药物指纹图谱进行对照分析,比较药物出峰情况,从化学成分角度分析贞芪扶正胶囊及其中药配方颗粒、传统饮片所含有效成分是否具有一致性。结果贞芪扶正胶囊对照品中特女贞苷、毛蕊异黄酮葡萄糖苷、黄芪甲苷在各自范围内线性关系良好(r=0.9998、0.9996、0.9997)。重复性试验和稳定性试验表明3种剂型供试品各成分重复性良好且在24 h内稳定。加样回收率试验表明HPLC对3种剂型供试品各成分检测的准确度良好。毛蕊异黄酮葡萄糖苷、特女贞苷、黄芪甲苷在贞芪扶正胶囊中含量分别为(5.384±0858)、(25849±3142)、(36065±3026)μg/mL,在中药配方颗粒中含量分别为(5825±0965)、(25463±3075)、(35618±2847)μg/mL,在传统饮片中含量分别为(5564±0958)、(25974±2986)、(35459±2468)μg/mL;3种剂型中上述有效成分的含量均无差异(P>0.05)。结论贞芪扶正胶囊及其中药配方颗粒、传统饮片主要有效成分相同,含量无差异,为贞芪扶正胶囊及中药配方颗粒在临床上替代传统饮片提供了依据。

Sources of varieties and quality of circular Fructus Ligustri Lucidi

This study aimed to trace sources and quantitatively analyze the specnuezhenide content of circular Fructus Ligustri Lucidi for clinical use. Different specifications of Fructus Ligustri Lucidi were identified using DNA barcoding technology and the specnuezhenide content was analyzed by High Performance Liquid Chromatography(HPLC). The ITS sequence of circular Fructus Ligustri Lucidi was identical to that of standard privet, which was determined through botanical identification. ITS sequence similarity between circular Fructus Ligustri Lucidi and Fructus Ligustri Lucidi which was registered in NCBI ranged from 99.5% to 100%. The sequences of circular and other Fructus Ligustri Lucidi were clustered in a Neighbor-Joining tree with bootstrap value of 95, and these sequences could be distinguished from adulterants. Conforming to pharmacopoeia standard, the average specnuezhenide content of circular Fructus Ligustri Lucidi was higher than that of chicken waist Fructus Ligustri Lucidi. Circular Fructus Ligustri Lucidi derived from Ligustrum lucidum Ait. and the specnuezhenide content was higher in circular Fructus Ligustri Lucidi than that in chicken waist Fructus Ligustri Lucidi.

碱水解法提取女贞子中红景天苷的工艺研究

目的:优化碱水解法提取女贞子中红景天苷的工艺条件。方法:按照《中华人民共和国药典》中分析方法验证指导原则对含量测定方法进行方法学验证,再通过单因素试验和正交试验对碱水解提取工艺进行优化。结果:最佳碱水解提取工艺参数为提取时间2 h、液固比40∶1(mL/g)、40%乙醇、1.0%NaOH,碱水解后红景天苷提取率达到1.80%,是未经碱水解提取率的8.5倍。结论:该法操作简单、提取率高、环境污染小,为以红景天苷为主成分的女贞子提取物的开发利用提供了理论依据。