Late Pliocene diversity and distribution of Drynaria(Polypodiaceae) in western Yunnan explained by forest vegetation and humid climates

The palaeodiversity of flowering plants in Yunnan has been extensively interpreted from both a molecular and fossil perspective. However, for cryptogamic plants such as ferns, the palaeodiversity remains poorly known. In this study, we describe a new ferny fossil taxon, Drynaria lanpingensis sp. nov. Huang,Su et Zhou(Polypodiaceae), from the late Pliocene of northwestern Yunnan based on fragmentary frond and pinna with in situ spores. The frond is pinnatifid and the pinnae are entirely margined. The sori are arranged in one row on each side of the primary vein. The spores have a semicircular to bean-shaped equatorial view and a tuberculate surface. Taken together with previously described fossils, there are now representatives of three known fossil taxa of Drynaria from the late Pliocene of western Yunnan.These finds suggest that Drynaria diversity was considerable in the region at that time. As Drynaria is a shade-tolerant plant, growing preferably in wet conditions in the understory of forests, its extensive existence may indicate forest vegetation and humid climates in western Yunnan during the late Pliocene.This is in line with results from floristic investigations and palaeoclimatic reconstructions based on fossil floras.

Effect of Drynaria total flavonoids on the expression of NMDAR1,GluR2 and CaMKⅡin the brain of hydrocortisone model mice

Objective:To study the effect of total flavonoids of Rhizoma Drynariae on the expression of NMDAR1,GluR2 and CAMKⅡprotein in model mice of kidney deficiency induced by exogenous glucocorticoid hydrocortisone and its mechanism.Methods:Kunming(KM)mice were randomly divided into the blank group,the hydrocortisone model group,the anti-brain failure capsule group,the Drynaria total flavonoids group,the Drynaria total flavonoids+ER blocker group,with 15 animals in each group.Except for the blank group,all groups were injected intramuscularly with hydrocortisone(25 mg·kg^(-1)·d^(-1))to create models.The water maze experiment,new object recognition experiment and platform jump experiment were used to conduct behavioral investigations.HE staining was used to observe the pathological changes of mouse hippocampus,and Western blotting detected the expressions of NMDR1,GluR2 and CAMKⅡproteins in the hippocampus of mice in each group.Results:The experimental results showed that compared with the model group,the learning and memory ability of the mice in the Drynaria fortunei total flavonoids group was significantly improved,and the difference was statistically significant(P<0.01),the expression of NMDR1 and GluR2 proteins in the hippocampus of the mice was significantly increased(P<0.01),and the level of CAMKⅡprotein significantly decreased(P<0.01).Conclusion:The total flavonoids of Rhizoma Drynariae may enhance the expression of NMDAR1 and GluR2 protein in the brain of hydrocortisone model mice through ER,reduce the expression of CAMKⅡprotein,and alleviate the damage to the brain tissue of the model mice and play a neuroprotective effect.

Determination of Naringin Content in Rhizoma Drynariae by High Performance Liquid Chromatography

[Objectives]To accurately determine the naringin content in Rhizoma Drynariae.[Methods]The high performance liquid chromatography(HPLC)method was applied in the determination of the naringin content in Rhizoma Drynariae.The sample was sonicated at room temperature.The mobile phase was 0.1%phosphoric acid-acetonitrile(75∶25),detected by diode array detector at the wavelength of 284 nm,and quantified by external standard method.[Results]The linearity of naringin was good in the concentration range of 5-500μg/mL with a correlation coefficient of 0.9999.[Conclusions]This method has good linearity,easy operation,correctness and reproducibility as required,and is expected to provide a method for the determination of naringin content in Rhizoma Drynariae.

Network Pharmacological Mechanism Research of Herba Drynariae Rhizoma-Epimedii Folium in Treating Osteoarthritis

Objective: To investigate the potential mechanism of Drynariae Rhizoma-Epimedii Folium in the treatment of osteoarthritis (OA) based on network pharmacology. Methods: The potential active constituents and targets of Drynariae Rhizoma-Epimedii Folium were screened through the traditional Chinese medicine (TCM) systems pharmacology database and analysis platform (TCMSP). Genecards database is used to find relevant targets of OA. The targets of “Drynariae Rhizoma-Epimedii Folium” were mapped to the targets of OA, and used Cytoscape software to build a “drug-ingredient-target-di- sease” regulatory network and protein protein interaction (PPI) network. R software was used to analyze the Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment of traditional Chinese medicine-disease targets. Results: Thirty-four effective ingredients and 130 traditional Chinese medicine-disease targets were screened out for the treatment of OA. The GO functions of traditional Chinese medicine-disease targets mainly included cytokine activity, cytokine receptor binding, nuclear receptor activity, transcription factor activity, proximal promoter DNA-binding transcription activator activity, DNA-binding transcription activator activity, phosphatase binding and so on. KEGG pathways involved in traditional Chinese medicine-disease targets mainly included TLR4 signaling pathway, TNF signaling pathway, IL-17 signaling pathway, MAPK signaling pathway, PI3K/AKT signaling pathway, apoptotic signaling pathway and so on. Conclusion: Network pharmacology may predict the multiple targets and multiple signaling pathways in Drynariae Rhizoma-Epimedii Folium treatment for OA, providing new ideas for future research.

Rhizome drynariae extract regulates osteoblast viability and proliferation as well as related gene expression: an experimental study

Objective:To study the effect of rhizoma drynariae extract on osteoblast viability and proliferation as well as related gene expression. Methods:SD rats were selected, and the osteoblasts in the femur were isolated, cultured and divided into the rhizoma drynariae group that were treated with 0.02 g/mL rhizoma drynariae extract and the control group that were treated with serum-free medium;24 h, 36 h and 48 h after treatment, CCK-8 kits were used to determine osteoblast proliferation activity, and fluorescent quantitative PCR kits were used to determine the mRNA expression of osteogenic activity-related genes and proliferation-related genes.Results:24 h, 36 h and 48h after treatment, osteoblast proliferation activity of rhizoma drynariae group was significantly higher than that of control group;Runx2, OPN, OCN, ALP, OPG, PI3K, AKT, EKR1/2, CyclinD1, CDK2, CDK4 and E2F mRNA expression in osteoblasts of rhizoma drynariae group were significantly higher than those of control group. Conclusion:Rhizoma drynariae extract can promote osteoblast proliferation, increase the osteogenetic activity of osteoblasts and also accelerate the cell cycle process through the PI3K/AKT and ERK1/2 signaling pathways.

药用蕨类植物骨碎补(Drynaria fortune)根茎繁殖研究

以中药材骨碎补基源植物槲蕨为试材,应用多重比较与主成分分析方法考察不同基质、激素类型及其浓度与处理时间对骨碎补根茎生根率、生根数、根长、叶片数、新茎数、新茎长与根系效果指数的影响。结果表明:以砂石或砂石∶壤土=1∶1繁殖基质,可有效提高骨碎补插穗生根率和生根数;当基质为砂石∶壤土=1∶1时更有利于根系的生长,提高根系效果指数、促进骨碎补插穗生长。生长调节剂在一定浓度范围内对骨碎补生长具有一定的促进作用,但高于一定浓度呈抑制趋势。主成分分析结果表明,浓度为100 mg·L-1处理综合得分最高,处理1.5~3.0 h可显著提高骨碎补插穗根系效果指数,促进骨碎补根系发育与茎叶分化。该研究提供了促进骨碎补根茎根系发育与后期生长方法,可为骨碎补的人工繁殖提供参考。

Interventional Value of Total Flavonoids from Rhizoma Drynariae on Cathepsin K,a Potential Target of Osteoporosis

Osteoporosis,the sixth most common disease in the world,is bringing increasingly serious harm to people＇s health.Cathepsin K,which plays an important role in bone resorption,is a potential target in the treatment of osteoporosis.Total flavonoids,the active ingredients in Rhizoma Drynariae,have shown obvious therapeutic effect on osteoporosis.In previous studies,it was presumed that the mechanism for the therapeutic effect was through inhibiting the expression of Cathepsin K.However,there are still no detailed reports on some key issues such as the specific inhibitory results of total flavonoids on Cathepsin K and the pathway of inhibition and so on.Based on previous studies on total flavonoids from Rhizoma Drynariae,the pathway for the effect of total flavonoids inhibiting Cathepsin K and their interventional value on Cathepsin K were analyzed in this paper, so as to explore the interventional feasibility and value of total flavonoids in Rhizoma Drynariae on Cathepsin K.

Population Pharmacokinetics of Naringin in Total Flavonoids of Drynaria Fortunei(Kunze) J.Sm.in Chinese Women with Primary Osteoporosis

Objective： To evaluate the effect of covariates on the pharmacokinetic profiles of naringin in the total flavonoids of Drynaria fortunei （Kunze） J. Sin. in the Qianggu Capsule （强骨胶囊） by evaluating Chinese women with primary osteoporosis. Methods： A total of 98 female patients from the communities of Jingshan, Beixinqiao, Jiaodaokou, Chaoyangmen, and Donghuamen in Beijing, China, aged 40 to 80 years, were included in this study. Blood samples were collected before and 0.5, 1, 2, 3, 4, 6, 8, 10, 12, and 24 h after a single oral dose of Qianggu Capsule. The concentration in blood samples from 32 patients before and 0.5, 1, 2, 3, and 4 h after drug administration were determined by liquid chromatography-tandem mass spectrometry （LC-MS/MS） method, and full set of pharmacokinetic data was analyzed with nonlinear mixed-effect modeling （NONMEM） software. The mean of population parameters clearance （C1）, central distribution volume （V）, absorption rate constant （Kal）, inter-compartmental clearance （C2）, peripheral distribution volume （V2） were set as parameters and estimated through base model, covariate model, and final model. Age, height, weight, blood urea nitrogen （BUN）, serum creatinine （Scr）, alanine transaminase （ALT）, aspartate transaminase （AST）, hyperlipidemia, Liver （Gan） Kidney （Shen） yin insufficiency （GSYI）, Kidney （Shen） yang insufficiency （SYI） were set as covariates. Results： The relationships between these parameters and covariates were analyzed. The results showed that C1 was the main parameter influenced by the selected covariates among the population parameters, and the relationships between the covariates and C1 were analyzed, among the selected covariates hyperlipidemia was identified as significant covariate of C1. Conclusion： The pharmacokinetic behaviors of naringin are altered with hyperlipidemia in Chinese women with primary osteoporosis.

Rhizoma Drynariae Improves Endometrial Receptivity in a Mus Model of Dysfunctional Embryo Implantation

Background: Rhizoma drynariae is a traditional Chinese medicine used in orthopedics and traumatology, but its effect on endometrial receptivity remains unknown. Aims and Objectives: To observe effect of Rhizoma drynariae and its main components on endometrial receptivity in a mus model of dysfunctional embryo implantation. Materials and Methods: Mus models were established by the Gn RHa+HMG+HCG method. Normal mus receiving saline were used as controls and the remaining six groups were: model receiving saline, progynova, aspirin, Rhizoma drynariae, osteopractic total flavone, and naringin. Pinopodes in uterine endometrium were examined by scanning electron microscopy. Stem cell factor(SCF) m RNA expressions was determined by real-time RT-PCR, and estrogen receptor α(ERα), progesterone receptor(PR) by immunohistochemistry. Results: In the model group, surface morphology of endometrium was heterogeneous, without obvious pinopodes. In the Rhizoma drynaria and progynova groups, pinopodes were abundant. Compared with the blank group, model group had lower levels of SCF(-47%), ERα(-63%) and PR(-50%)(all P<0.05). In comparison, Rhizoma drynariae group had higher levels of SCF(+73%), ERα(+118%) and PR(+101%)(all P<0.01). The individual main components of Rhizoma drynariae had variable efficacy. Conclusion: Rhizoma drynariae could improve endometrial receptivity of mouse models of dysfunctional embryo implantation as shown by increased numbers of pinopodes and higher levers of SCF, ERα, PR compared with the model group.

骨碎补大棚栽培品与野生品的广靶代谢组学比较分析

【目的】骨碎补是一种多年生传统中药材,目前药材原料多来源于野生采集,鲜见有人工栽培的报道。因过度采集,骨碎补野生资源破坏严重,导致药材资源锐减,人工种植成为必然趋势。在探索骨碎补设施大棚栽培技术的基础上,利用代谢组学技术开展大棚栽培药材与野生药材的靶向代谢组学比较研究,探讨栽培品与野生品主要代谢产物的差异,为骨碎补人工栽培技术推广提供依据。【方法】以骨碎补基源植物槲蕨为研究对象,采用UPLC-MS/MS分析技术对比分析设施大棚栽培3年的槲蕨根茎与其野生根茎之间的差异代谢物,并开展差异代谢物富集通路分析。【结果】从两种药材检测到的749种代谢物中筛选到100种差异代谢物,且与野生品相比,大棚栽培品中有58种代谢物含量上调、42种含量下调。这些差异代谢物主要包括黄酮类化合物、有机酸、氨基酸及其衍生物、酚酸类、生物碱、游离脂肪酸等。在大棚栽培品中,黄酮醇、黄酮碳糖苷、糖及糖类代谢物含量全部上调;而在野生品中,氨基酸及衍生物、酚酸类、溶血磷脂酰胆碱和花青素类代谢物含量上调幅度较大。通路富集分析结果显示,100个差异代谢物共注释到47条代谢通路,其中显著富集的有维生素B6代谢、芪类化合物生物合成、二芳基庚烷生物合成、姜酚的生物合成、黄酮和黄酮醇生物合成、半胱氨酸代谢、蛋氨酸代谢、花青素生物合成、氨酰基tRNA生物合成等途径。【结论】大棚栽培的骨碎补,其黄酮醇、黄酮碳糖苷、糖及糖类代谢物化合物含量显著高于野生品,因总黄酮被公认为主要药效成分,因此大棚栽培的骨碎补比野生品具有主效成分优势,可为骨碎补规模化设施栽培或人工栽培提供科学依据。

骨碎补及其活性成分防治骨质疏松症的作用机制

骨碎补是补肾强骨的经典药材,药理学研究显示其主要活性成分为黄酮类和二氢黄酮类柚皮苷,其在抗骨质疏松、修复骨缺损等方面发挥着积极作用。细胞分子生物学研究提示骨碎补对成骨细胞、破骨细胞和间充质干细胞具有多通路、多因子调节作用,如Wnt/β-catenin、MAPK、OPG/RANKL/RANK、Notch、BMP-Smads、PI3K/Akt等信号通路和TNF-α、白介素-1(IL-1)、白介素-6(IL-6)、雌激素及其受体、组织蛋白酶K(CTSK)等细胞因子。笔者重点综述了中药骨碎补及其提取物防治骨质疏松的信号通路和细胞因子研究进展,旨在为骨碎补的药理药效研究及临床应用提供参考。

骨碎补总黄酮及其成分柚皮苷促进牵张成骨新骨愈合作用比较

目的中药骨碎补是中医骨伤科的常用药,现代研究表明,骨碎补总黄酮及其成分柚皮苷均能促进牵张成骨过程。通过动物体内实验,观察比较骨碎补总黄酮和柚皮苷对牵张成骨骨愈合的影响。方法取雄性新西兰兔15只,按前期研究造模步骤完成牵张成骨模型兔造模;将模型兔分为3组(n=5),在术后第2天开始分别予生理盐水、骨碎补总黄酮[200 mg/(kg·d)]和柚皮苷[100 mg/(kg·d)]灌胃给药。术后第56天处死动物,观察各组骨标本牵张区骨缺损修复情况,并行显微CT(Micro computed tomography,Micro-CT)检查、生物力学检测。结果与柚皮苷组相比,骨碎补总黄酮组骨骼矿物质密度(Bone mineral density,BMD)、骨表面积组织体积比值(Bone volume to tissue volume,BV/TV)、骨小梁厚度(Trabecular thickness,Tb.Th)、骨小梁数量(Trabecular number,Tb.N)更高,差异有统计学意义(P<0.05),而骨碎补总黄酮组骨小梁分离度(Trabecular separation,Tb.Sp)更小,差异有统计学意义(P<0.05)。与柚皮苷组相比,骨碎补总黄酮组骨标本能承受的最大应力(Maximum stress,N/mm^(2))和最大载荷(Maximum loading,N)均更大(P<0.05)。结论本研究表明,骨碎补总黄酮和柚皮苷对牵张成骨骨愈合均有促进作用,而总黄酮比其成分柚皮苷更显著。

中药复方治疗原发性骨质疏松用药规律及作用机制分析

背景:中药复方治疗原发性骨质疏松有着较久的历史,并且疗效确切,但其用药规律及作用机制尚不明确。目的:利用数据挖掘与网络药理学的方法学,探究现代中医家治疗原发性骨质疏松的用药规律、分子层面的作用机制并一定程度的加以验证。方法:将中国知网、万方、维普、PubMed等数据库收录的相关文献作为数据来源,使用Microsoft EXCEL 2019、IBM SPSS 25.0等软件统计和提取相关资料。将数据统计得到的高频药物进行关联规则分析和聚类分析,综合两者结果得到中药复方治疗原发性骨质疏松的核心药物组合,利用网络药理学阐释此组合发挥治疗作用的机制并借助分子对接加以验证。结果与结论:最终纳入151篇文献,筛选出方剂207首,涉及中药285味。①通过上述两种分析综合得到重要药物组合10组,其中置信度和提升度最高的核心药物组合为“骨碎补-杜仲-当归”,其组合治疗原发性骨质疏松的关键成分为槲皮素、山奈酚、柚皮素等,核心靶点为SRC原癌基因(SRC proto-oncogene)、磷脂酰肌醇-3-激酶调节亚单位1(PIK3R1)、RELA原癌基因(RELA)等,主要通路为癌症信号通路、JAK-STAT信号通路、VEGF信号通路、核因子κB信号通路等。②将关键活性成分与核心靶点进行分子对接,两者展示出较好的结合性。③中药复方治疗原发性骨质疏松核心药物组合可利用其多种活性成分,通过多条信号通路、作用于多个靶点发挥疗效,可为后续治疗原发性骨质疏松新药的研发提供理论基础。

骨碎补活性成分促进骨缺损后骨重建的机制及其组织工程学应用

骨缺损的治疗难度大、周期长,一直都是骨科临床面临的重大挑战。骨碎补是我国中医骨伤科的常用药材,其活性成分(主要为黄酮类)可促进骨髓间充质干细胞成骨分化、成骨细胞增殖、成血管-成骨耦联,抑制破骨细胞活力,从而促进骨缺损区的骨质矿化和修复重建。骨碎补活性成分是骨再生治疗药物的良好替代品,将其负载于组织工程支架材料上,可极大地提高药物的生物利用度。同时,缓释微球进一步解决了支架药物的突发性释放等问题,应用其所制备的含骨碎补活性成分的复合支架具有较好的成骨活性和骨诱导性,骨修复效果确切,可满足骨移植物的多样化性能要求,临床应用前景广阔。

骨碎补抑制骨髓腔内脂肪异常积累引起的脂毒性介导的成骨细胞焦亡及其机制

目的探讨骨碎补(rhizoma drynariae,Rhd)对骨髓腔内脂肪异常积累引起的脂毒性介导成骨细胞焦亡的影响及其机制。方法体内构建卵巢摘除(ovariectomy,OVX)骨质疏松小鼠模型,用Rhd进行灌胃,给药8周后称重,取股骨组织和血清。HE染色观察髓腔内脂肪堆积状态。体外构建成脂-成骨细胞共培养系统,茜素红S(alizarin red S,ARS)染色观察钙化结节的数量,碱性磷酸酶(alkaline phosphatease,ALP)染色检测碱性磷酸酶活性水平,乳酸脱氢酶(lactate dehydrogenase,LDH)检测和Hoechst33342/PI染色检测共培养下成骨细胞(osteoblasts,OBs)焦亡水平,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞培养上清液中白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-18(interleukin-18,IL-18)的释放量,Western blot检测成骨能力相关蛋白Runt相关转录因子2(runt-related transcription factor 2,RUNX2)、骨形态发生蛋白2(bone morphogenetic protein,BMP2)和1型胶原蛋白(collagen-1,COL-1)。选择NLRP3特异性抑制剂MCC950作为阳性对照检测焦亡相关蛋白NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、半胱天冬酶-1(cysteinyl aspartate specific proteinase-1,CASP-1)、效应蛋白消皮素D(gasdermin D,GSDMD)、IL-1β和IL-18的表达。结果OVX小鼠髓腔内发生骨质流失伴大量脂肪堆积,Rhd灌胃可以有效缓解这一趋势。在成脂-成骨共培养系统中,共培养环境抑制了OBs活性和OBs中ALP活性及矿化结节,抑制了RUNX2、BMP2、COL-1蛋白表达,并且诱导焦亡发生,提高了LDH释放量和PI染色细胞阳性率,以及上调NLRP3、CASP-1、GSDMD、IL-1β和IL-18蛋白表达水平。而这一趋势在Rhd干预后得到逆转。结论大量脂肪堆积引起的脂毒性可以抑制OBs活性并通过激活NLRP3炎症小体诱导OBs焦亡,而Rhd可能通过抑制NLRP3炎症小体的激活以抑制OBs焦亡和炎性反应,从而起到抗骨质疏松的作用。

骨碎补总黄酮调控H型血管影响大鼠股骨Masquelet诱导膜模型的骨重建

背景:多项研究发现,骨碎补总黄酮可促进诱导膜中血管新生、改善诱导膜生物性能、加速诱导膜技术骨重建,但相关分子机制仍需进一步探究。目的:观察骨碎补总黄酮通过调控H型血管对大鼠股骨Masquelet诱导膜模型骨重建的影响。方法:将36只雄性SD大鼠按体质量分层后随机分为空白组、模型组、中药组,每组12只,3组均建立4 mm右后肢股骨骨缺损模型,模型组与中药组骨缺损处充填聚甲基丙烯酸甲酯骨水泥;造模后6周,空白组骨缺损处填充大鼠自体尾骨,模型组与中药组取出诱导膜内的骨水泥后植入大鼠自体尾骨。植骨第3天开始,中药组灌胃给予骨碎补总黄酮157.5 mg/(kg·d),其余两组灌胃给予生理盐水,连续给药至植骨后8周。植骨后8周取材进行相关检测。结果与结论:①X射线片显示,空白组缺损区骨折线清晰,仅有少量骨痂形成;模型组缺损区可见不连续皮质骨,骨缺损区仍存在;中药组缺损区充满新生骨组织,骨髓腔与部分皮质骨形成,骨折线消失。②Micro-CT扫描显示,空白组缺损区新生骨量较少,模型组缺损区骨小梁数量明显增多,中药组大量新生骨组织填充于骨缺损区。③苏木精-伊红染色显示,空白组缺损区仅见少量新骨形成,成骨质量较差;模型组缺损区有较多的新骨形成,但骨组织内夹杂有部分纤维结缔组织;中药组缺损区可见大量新骨形成,成骨质量最佳。④CD31/Emcn免疫荧光双标染色显示,空白组骨缺损区新生骨组织内H型血管数量稀少、分布稀疏;相比空白组,模型组骨缺损区骨组织内含更多H型血管,血管呈相对规则的条状分布;中药组骨缺损区H型血管数量最多,并且血管分布密集。⑤结果表明,骨碎补总黄酮可通过上调H型血管表达增强成血管-成骨作用,提高大鼠股骨Masquelet诱导膜模型成骨效能、促进骨重建。

基于网络药理学和分子对接探讨三七-骨碎补治疗股骨头坏死的作用机制

目的 基于网络药理学和分子对接探究“三七-骨碎补”药对治疗股骨头坏死的作用机制。方法 利用中药系统药理学数据库与分析平台(TCMSP)对“三七-骨碎补”药对的活性成分及成分靶点进行采集、筛选,通过CNKI、PubMed数据库对其进行补充,并利用UNIPORT数据库对靶点进行转换;通过GeneCards、TTD和OMIM数据库收集股骨头坏死的作用靶点,取得交集靶点后导入STRING数据库,生成PPI网络,并通过Cytoscape 3.9.0软件构建中药-活性成分-靶点网络;通过DAVID数据库对核心靶点进行GO功能富集分析与KEGG通路富集分析。结果 共得到“三七-骨碎补”药对的20个活性成分和229个潜在靶点,1386个股骨头坏死的靶点。GO富集分析主要包括炎症反应、凋亡过程、调节血管生成、与雌性激素响应等功能;KEGG富集分析主要包括脂质与动脉硬化、白细胞介素-17信号通路、类风湿性关节炎等信号通路。结论 “三七-骨碎补”药对治疗股骨头坏死具有成分多、靶点广、通路复杂的特点,可能通过IL6、TNF、TP53等靶点以及脂质代谢、白细胞介素-17等信号通路共同发挥治疗股骨头坏死的作用。

槐黄烷酮G抑制破骨细胞分化和减少骨丢失的机制研究

目的 通过体内与体外实验相结合探讨槐黄烷酮G抑制破骨细胞形成和分化,从而缓解骨丢失现象的作用机制研究。方法 通过CCK-8和TRAcP染色实验验证槐黄烷酮G是否具有抑制破骨细胞形成的作用;通过分子对接验证槐黄烷酮G与破骨细胞分化相关的特异性蛋白结合的亲和力大小,借助RT-PCR和Western blot实验探讨槐黄烷酮G抑制破骨细胞形成的作用机制;最后通过体内实验进一步探讨,以验证体外实验的准确性和可靠性。结果 槐黄烷酮G能够在安全浓度下抑制破骨细胞形成和分化,同时也与破骨细胞相关的特异性蛋白如NFATc1、CTSK、c-Fos、MMP9具有较高的亲和力,并能够降低上述基因和蛋白的相对表达水平,槐黄烷酮G也能够在15和30 min处有效降低MAPK信号通路中的ERK和JNK的磷酸化表达水平,体内实验证明槐黄烷酮G能够起到缓解骨丢失,改善骨小梁相关参数的作用。结论 槐黄烷酮G具有抑制破骨细胞形成和分化,缓解骨量减少的作用,从而起到抗骨质疏松症的作用。

骨碎补UPLC指纹图谱与人骨肉瘤细胞MG63中碱性磷酸酶活性的谱效关系研究

建立骨碎补生品和砂烫品提取物的UPLC指纹图谱,并研究骨碎补提取物与人骨肉瘤细胞MG63中碱性磷酸酶(ALP)活性之间的谱效关系。采用UPLC法建立骨碎补生品和烫品的指纹图谱并通过UPLC-Q-TOF MS法对其化学成分进行鉴别;通过测定MG63细胞中ALP活性检测骨碎补提取物对MG63细胞成骨分化的影响;采用灰色关联度分析、双变量相关性分析以及偏最小二乘回归分析法分析骨碎补生品和烫品的UPLC特征指纹峰与其促ALP活性之间的谱效关系。在标定的12个共有峰中,确定了4、5、12号峰与ALP活性呈正相关,其对应的物质依次是咖啡酸-4-O-β-D-吡喃葡萄糖苷、儿茶素7-葡糖苷、柚皮苷,为深入研究骨碎补的促成骨分化活性物质基础提供依据。

态靶辨证在风寒湿痹型类风湿关节炎中的应用

类风湿关节炎是临床常见疾病,易致关节畸形甚至功能丧失。在临床实践中,发现蠲痹汤加味对风寒湿痹型类风湿关节炎疗效尚可。“态靶辨证”是仝小林院士通过多年临床经验总结得出,基于“态靶辨证”理论,论述蠲痹汤加骨碎补调风寒湿痹型类风湿关节炎“寒”态以及肝肾“虚”态,运用羌活+独活药对治疗其靶症,态靶双调。