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.

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。

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。

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。

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。

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。

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。

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。

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。

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。

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.

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.

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.

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.

女贞子治疗去卵巢大鼠骨质疏松的实验研究

目的观察女贞子对去卵巢(ovariectomy,OVX)大鼠骨质疏松(osteoporosis,OP)的治疗作用,并探讨其作用机制。方法大鼠骨质疏松模型,女贞子(9、4.5、2.25 g.kg-1.d-1)灌胃给药,实验过程称体重,连续给药26周后测定血钙(S-Ca)、血磷(S-P)、尿钙(U-Ca)、尿磷(U-P)的含量,ELISA试剂盒双抗体夹心法测定血清碱性磷酸酶(ALP)、血清骨特异性碱性磷酸酶(BALP)含量、骨钙素(OCN)含量、尿液中脱氧吡啶啉(DPD)含量,DEXA型骨密度仪分析大鼠的股骨、胫骨、第4椎骨的BMD(bone mineral density),动物处死后完整取下心、肝、脾、肺、肾、胸腺、脑、子宫进行称重,采用HE染色法进行子宫病理学检查。结果女贞子能有效抑制去卵巢所致的大鼠体重增加、升高S-Ca、S-P、降低U-Ca、U-P、降低ALP、BALP、OCN、DPD的含量,增加大鼠股骨、胫骨、第4椎骨的BMD,并且长期用药对子宫无明显刺激作用。结论女贞子对OVX大鼠所致的骨质疏松症有良好的治疗效果,其作用机制可能与其能降低高的骨转换率有关。

女贞子的药理作用及其临床应用进展

女贞子具有抗肿瘤、护肝、调节免疫功能、延缓衰老、抗炎和降血脂等多种药理作用。在肝炎、内分泌代谢病和反复呼吸道感染等多种疾病的临床治疗中,女贞子具有广泛的应用前景。本文通过查阅国内外相关文献,对女贞子的药理作用及其临床应用进展进行综述。

大孔吸附树脂纯化女贞子中齐墩果酸和熊果酸的研究

目的研究大孔吸附树脂纯化女贞子中齐墩果酸和熊果酸的动态吸附与解吸工艺,为工业化生产提供参考。方法采用高效液相色谱法测定齐墩果酸和熊果酸的量;运用静态吸附与解吸试验对树脂型号进行筛选,结合单因素试验优选女贞子齐墩果酸和熊果酸的纯化操作条件。结果HPD-100树脂对齐墩果酸和熊果酸的吸附与解吸性能较好;吸附的优化条件为树脂床径高比1:6,料液质量浓度0.20g(生药)/mL,体积流量2BV/h和吸附容量2.4mL/g(树脂);最佳解吸条件为乙醇体积分数80%,体积流量2BV/h和溶剂用量4BV;产品中齐墩果酸、熊果酸的质量分数分别为25.21%、15.65%,转移率分别达98.64%、98.58%。结论HPD-100可有效地富集、纯化女贞子中齐墩果酸和熊果酸。

女贞子的抗炎作用

女贞子对二甲苯引起小鼠耳廓肿胀、乙酸引起的小鼠腹腔毛细血管通透性增加及对角叉菜胶、蛋清、甲醛性大鼠足垫肿胀均有明显抑制作用,并显著降低大鼠炎症组织PGE的释放量。女贞子抑制大鼠棉球肉芽组织增生,同时伴有肾上腺重量的增加,对大鼠胸腺重量无明显影响。

女贞子中三萜化合物的提取与含量测定

以超声浓方法提取女贞子中三萜化合物,以紫外光谱法检测其含量。通过单因素试验和正交试验考察了超声处理时间、液固比、乙醇浓度三因素对超声波提取三萜化合物的影响。结果表明,最佳提取工艺条件为超声时间30min、液固比(V/W)8:1、无水乙醇。在此条件下测得三萜化合物的含量为5.61%(W/W)。