Sulfated Cyclocarya paliurus polysaccharides exert immunomodulatory potential on macrophages via Toll-like receptor 4 mediated MAPK/NF-κB signaling pathways

The biological activity of plant polysaccharides can be enhanced by sulfated modification.In this study,the immunomodulatory effect of sulfated Cyclocarya paliurus polysaccharides(SCP3)on macrophages RAW264.7 and its potential molecular mechanism were investigated.Results showed that SCP3 at 25-100μg/m L increased viability and improved phagocytosis of RAW264.7 cells.Meanwhile,SCP3 could activate mitogen-activated protein kinase(MAPK)and nuclear factor kappa B(NF-κB)signaling pathways,which increased the phosphorylation of Erk1/2,JNK,p38 and NF-κB p65,promoting secretion of cytokines tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and nitric oxide(NO)as well as the production of reactive oxygen species(ROS).In addition,Toll-like receptor 4(TLR4)receptor inhibitors were able to block the production of NO and TNF-αby SCP3-stimulated macrophages.Based on Western blot analysis and validation using specific inhibitors against MAPK and NF-κB signaling pathways,the results demonstrated that SCP3 induced macrophages activation and enhanced TNF-αand NO production via TLR4-mediated MAPK and NF-κB pathways.In summary,SCP3 has significant immunomodulatory potential.The underlying molecular mechanism was that SCP3 activates macrophages via TLR4 receptors to promote ROS production,which in turn activates the downstream MAPK/NF-κB signaling pathway and then increases the secretion levels of cytokines and NO.

Apoptosis of colon cancer CT-26 cells induced polysaccharide from Cyclocarya paliurus and its phosphorylated derivative via intrinsic mitochondrial passway

In this study,the antitumor properties and the possible molecular mechanisms of Cyclocarya paliurus polysaccharide(CP)and its phosphorylated derivative(P-CP)on CT-26 mouse colon carcinoma cells were investigated.Results found that CP had high inhibition ratio against CT-26 cells.The flow cytometry results found that CP treatment could cause the intracellular acidification,arrest the cell cycle in the S phase and increase reactive oxygen species generation.Additionally,CP treatment triggered mitochondrial membrane potential depolarization and Ca^(2+)overloading,and broke down the balance of antioxidant system,Na^(+)/K^(+)-ATPase and Ca^(2+)-ATPase.Further analysis found CP induced cell apoptosis through improving the activities of caspase-3 and caspase-9,and increasing the level of cytochrome C.Furthermore,the comparative study of antitumor effect on CT-26 cells displayed that the phosphorylation enhanced antitumor activities of polysaccharides.These results suggest CP is a potential natural therapeutic agent for colon cancer and phosphorylation represents an effective method of enhancing the antitumor activity of CP.

Cytochemical localization of ATPase and sub-cellular variation in mesophyll cell of Cyclocarya paliurus seedlings under iso-osmotic stress and calcium regulation

The ultrastructural distribution and active location of ATPase and the ultrastructural variations were investigated in mesophyll cells of Cyclocarya paliurus seedlings after iso-osmotic salt/water treatments in combination with calcium regulation. C. paliurus seed- lings were treated with five groups （control, 85 mM NaCl, 85 mM NaCl＋ 12 mM Ca（NO3）2, PEG iso-osmotic to 85 mM NaCl and PEG iso-osmotic to 85 mM NaCl＋12 mM Ca（NO））2） in a hydroponic system in a phytotron. Results show that under normal growth conditions the ATPase activity was low and the enzyme was primarily located on the nucleus. After 12 days of iso-osmotic salt/water treatments, ATPase activity on the tonoptast increased. Osmiophilic globules for iso-osmotic water treatment were greater than that for iso-osmotie salt treatments. The ATPase activity increased and was mostly transferred onto the nucleus for calcium regulation treatment under iso- osmotic salt/water stresses, and the osmiophilic globules significantly decreased under iso-osmotic water stress with calcium regulation. The ATPase located on the nucleus indicated that the degree of salt/drought damage that seedlings suffered was slighter, while the amount of the enzyme located on the tonoplast showed that the degree of salt/drought damage there was more serious. After 4 and 20 days of iso- osmotic treatments, the injury suffered by the leaf ultrastructures of C. paliurus seedlings for iso-osmotic treatment with calcium regula- tion was lower than those without calcium regulation, especially for the iso-osmotic water treatments. Preliminary analysis suggests that the iniury suffered by C. paliurus seedlings was lower for iso-osmotic salt treatments than for iso-osmotic water treatments, while the effect of calcium regulation under iso-osmotic water stress was greater than that of the iso-osmotic salt stress.

Influence of genotypes and environmental factors on leaf triterpenoid content and growth of Cyclocarya paliurus

Cyclocarya paliurus, an economically valuable tree species, has traditionally been used as a nutraceutical food or medicine in China. However, limited information is available on its genotype selection and cultivation under a wide range of environmental conditions for growth and targeted health-promoting substances. We studied the effects of genotype and environment, and their interaction on leaf triterpenoid content and tree growth for 12 genotypes of C. paliurus grown at four sites. We quantified the correlation between leaf triterpenoid accumulation and tree growth. The contents of cyclocaric acid B, cyclocarioside I, and arjunolic acid ranged from 0.06 to 3.89, 0 to 3.71, and 0.65 to 8.86 mg g^-1, respectively. Three individual triterpenoids were primarily influenced by genotype (variation ranged from 53.7 to 68.0%), while environment accounted for most of the variation in total triterpenoid content and tree growth (71.3–89.5%). Most tested environmental parameters were significantly correlated to total triterpenoid content, but not to the contents of the individual triterpenoids measured. Growth in tree height and diameter at breast height were significantly negatively correlated with total triterpenoid content but were non-significantly correlated with individual triterpenoid contents. We conclude that genotypic selection, manipulation of environmental conditions, and implementation of appropriate silvicultural operations would be important strategies for increasing the accumulation of health-promoting phytochemicals.

Natural variations in flavonoids and triterpenoids of Cyclocarya paliurus leaves

Cyclocarya paliurus is widely distributed in subtropical areas of China.Secondary metabolites in the leaves,including flavonoids and triterpenoids,provide protection against diseases such as hyperlipidemia,diabetes,hypertension,and hypoimmunity.In this study,343 C.paliurus leaves were collected from 32 distinct populations covering most of the main distribution,to deduce the response of flavonoids and triterpenoids to seven environmental factors.Principal component analysis was performed using seven flavonoid and six triterpenoid compounds.The results show that altitude,latitude,and longitude significantly contributed to variations in total flavonoid and triterpenoid contents.In addition,the interaction of these metabolites was extensively correlated with environment changes,and therefore should be considered during selection and cultivation.

Protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis

OBJECTIVE To explore the protective effects of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja on STZ-stimulated INS-1 cells through regulating of autophagy and apoptosis.METHODS INS-1cells were cultured in media containing 3n M STZ and different doses of total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja.The proliferation of cells was examined by MTT assay,ROS content were detected by fluorescence enzyme label.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),hydrogen peroxidase(CAT),malondialdehyde(MDA)were also measured by colorimetry.The activities of caspase-3,9 were also observed by Caspase colorimetric assay kit in each group.The expressions of Bcl-2 m RNA and Bax m RNA were detected by Real time PCR,protein expression of LC3-Ⅱand PARP were detected by Western blotting.RESULTS Compared with the control group,total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja could promote the proliferation of INS-1 cells no matter with STZ or not when its concentration lower than 25μg·m L-1;but when its concentration higher than 100μg·m L-(1use individually)or 50μg·m L-1(combined use),total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja might significantly inhibite the growth of the cells whether STZ existed or not.Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja(6.25,12.5,25μg·m L-1)might inhibit INS-1cell apoptosis,decrease intra-cellular ROS contents,improve the s upernatant liquid SOD,GSH-PX,CAT activities,decrease MDA level,promote INS-1 cell secreting insulin,decrease the protein expressions of autophagy protein LC3Ⅱand apoptosis regulating protein cleaved PARP protein,up-regulate the anti-apoptotic protein Bcl-2 m RNA expression,down-regulate the pro-apoptotic protein Bax m RNA expression and Bcl-2and Bax,reduce the activity of caspase-9 and caspase-3(P<0.05 or P<0.01).CONCLUSION Total triterpenoids extracts from Cyclocarya paliurus(Batal.)Iljinskaja has good protective effect on STZ-stimulated INS-1cells.It can inhibit STZ injured INS-1 cells to overproduce ROS production,enhance endogenous antioxidant enzymes(GSH-Px,SOD,CAT activities),reduce the expression of autophagy protein LC3Ⅱand apoptosisregulating protein cleaved PARP,up-reglute the antiapoptosis protein Bcl-2 expression,down-regulate the pro-apoptotic protein Bax expression,decrease the caspase-9 and caspase-3 activities,and improve the INS-1cell survival rate,and then play a protective effect on damaged INS-1 cells.

Callus culture of Cyclocarya paliurus (Batal.) Iljinskaja leaves

The hormone combinations and dark culture of the callus induction for Cyclocarya paliurus （Batal.） Iljinskaja leaves were optimized by an orthogonal experiment. The effects of hormones on callus proliferation were also studied. The results show that the optimum medium for callus induction of C. paliurus leaves was MS ＋ 1.0 mg.L-1 6-BA ＋ 0.5 mg.L-1 NAA ＋ 1.0 mg·L-1 2,4-D. The combination of 6-BA and NAA significantly affected the induction of callus, while the effect of 2,4-D was less. The maximum rate of callus induction was 95.4%. A 10-day darkness period was beneficial for callus induction. The best proliferation medium was 1/4 DKW ＋ 0.5 mg·L-1 6-BA ＋ 1.0 mg·L-1 2,4-D.

Study on Response Surface Methodology Optimization for Extraction Process of Cyclocarya paliurus Extract and Its Blood Sugar-lowering Effect

[Objectives]This study was conducted to discuss the mechanism of the blood sugar-lowering effect of Cyclocarya paliurus extract.[Methods]With C.paliurus leaves as the raw material,the ethyl acetate-butyl acetate microwave extraction process of C.paliurus was studied.With the yield of C.paliurus extract as an evaluation index,single-factor experiments were carried out on such 6 factors as the dry C.paliurus leaf powder size,solid-to-liquid ratio,extraction time,microwave frequency,microwave power,and microwave extraction time,and the extraction and purification process was further optimized by response surface analysis.The normal mice and the hyperglycemic mice modeled by alloxan were injected with C.paliurus extract to study the blood sugar-lowering effects of different groups of C.paliurus extract.[Results]The optimal process combination for the extraction of C.paliurus leaves was obtained:dry C.paliurus powder size 90-100 mesh,solid-to-liquid ratio 1∶20(g/ml),extraction time 192 min,microwave frequency 2500 MHz,microwave power 490 W,and extraction time 248 s.Under these conditions,the yield of C.paliurus extract reached the maximum,95.10%,reaching 99.83%of the predicted value.The blood sugar-lowering test on mice showed that the C.paliurus extract had a good effect on lowering blood sugar,indicating that C.paliurus extract can improve the free radical scavenging capacity to a certain extent.[Conclusions]This study provides certain reference for the in-depth study of the physiological effects of C.paliurus extract and the comprehensive development and utilization of C.paliurus extract.

青钱柳苦荞膳食补充剂对大鼠的28天经口毒性试验

目的:研究青钱柳苦荞膳食补充剂对大鼠亚急性毒性。方法:将Wistar大鼠按体质量随机分为4组,即青钱柳苦荞膳食补充剂3个剂量组(1.67、3.33、6.67 g/kg)和1个常规基础饲料对照组。分别喂饲相应饲料28 d,自由进食饮水,观察大鼠体质量、食物利用率、血常规、血生化、尿液指标,大鼠麻醉后解剖进行大体观察、计算主要脏器的脏器系数并进行组织病理学检查。结果:实验组大鼠的体质量增长正常,未见明显毒性反应,其血常规、血生化、尿液指标、脏器系数与对照组比较差异均无统计学意义(均为P>0.05)。大鼠解剖时大体观察未发现明显异常。个别实验组大鼠心、肾脏质量与对照组比较差异具有统计学意义(P<0.05),病理组织学检查未发现青钱柳苦荞膳食补充剂引起的典型病理组织学改变和特异性损伤改变。结论:在本实验条件下,该青钱柳苦荞膳食补充剂经大鼠28天经口毒性试验未见明显的毒性作用。

青钱柳叶化学成分及其α-葡萄糖苷酶抑制活性研究

目的 研究青钱柳Cyclocarya paliurus(Batal.) Iljinskaja叶的化学成分及其α-葡萄糖苷酶抑制活性。方法 青钱柳叶95%乙醇提取物采用大孔树脂、硅胶、Sephadex LH-20、聚酰胺、C18反相硅胶及半制备HPLC进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。采用PNPG法评价其α-葡萄糖苷酶抑制活性。结果 从中分离得到15个化合物,分别鉴定为cyclopaloside C(1)、cyclopaloside A(2)、juglanosides E(3)、vaccinin A(4)、ent-mururin A(5)、山柰酚-3-O-α-L-鼠李糖苷(6)、山柰酚-3-O-β-D-葡萄糖苷(7)、山柰酚-3-O-β-D-葡萄糖醛酸甲酯(8)、山柰酚-3-O-β-D-葡萄糖醛酸乙酯(9)、山柰酚-3-O-β-D-葡萄糖醛酸丁酯(10)、槲皮素-3-O-α-L-鼠李糖苷(11)、槲皮素-3-O-β-D-葡萄糖苷(12)、槲皮素-3-O-β-D-半乳糖苷(13)、槲皮素-3-O-β-D-葡萄糖醛酸丁酯(14)、香橙素(15)。总提取物抑制α-葡萄糖苷酶的IC_(50)值为(1.83±0.04)μg/mL,化合物1、4~5分别为(29.48±1.86)、(0.50±0.07)、(0.71±0.07)μmol/L。结论 化合物1为新四氢萘醇苷类,化合物4~5、8~10、14为首次从青钱柳叶中分离得到。化合物4~5为较少见的黄酮木脂素类化合物,对α-葡萄糖苷酶具有潜在的抑制活性。

青钱柳多糖调节胰岛和肝脏葡萄糖转运蛋白4转位干预2型糖尿病大鼠的作用机制

目的 观察青钱柳多糖调节胰岛和肝脏葡萄糖转运蛋白4(GLUT4)转位改善2型糖尿病(T2DM)大鼠外周胰岛抵抗的作用。方法 建立T2DM大鼠模型(给予高脂饲料后注射链脲佐菌素35 mg·kg^(-1)),将造模成功的大鼠随机分为模型对照组,青钱柳多糖提取物小、大剂量组(5,10 g·kg^(-1))和盐酸二甲双胍组(0.25 g·kg^(-1)),每组9只,给药8周。测定空腹血糖、血脂变化;苏木精-伊红染色法观察胰岛和肝脏病理形态的改变;免疫组化法观察胰岛磷酸化磷酯酰肌醇3激酶(p-PI3K)、磷酸化丝氨酸苏氨酸蛋白激酶1(p-Akt1)、GLUT4蛋白的表达;免疫荧光观察肝脏和胰岛GLUT4转位。结果 与模型对照组比较,青钱柳多糖提取物小、大剂量组和盐酸二甲双胍组大鼠胰岛和肝脏结构较完整,血糖下降(P<0.05),高密度脂蛋白升高(P<0.05),胰岛p-PI3K、p-Akt1、GLUT4蛋白表达升高(P<0.05),肝脏和胰岛GLUT4转位增强(P<0.05)。结论 青钱柳多糖可调节T2DM大鼠糖脂紊乱,其机制可能是增强胰岛p-PI3K、p-Akt1、GLUT4蛋白的表达,促进肝脏和胰岛GLUT4转位,从而调节外周胰岛抵抗。

我国特有单种属珍稀资源植物青钱柳繁育技术研究进展

青钱柳是我国特有的单种属乔木及国家重点保护濒危植物之一,是极其重要的珍稀木本植物资源。近年来由于其具有降血糖、降血压、降血脂,增加机体免疫力、抗氧化、抗衰老等诸多功效,尤其是其具有修复受损胰岛β细胞作用的特性而受到国内外广泛关注。然而青钱柳种子败育严重并具有深休眠性,有性繁殖系数很低,其野生资源的稀缺和繁殖困难使这一资源的利用受到了极大的限制。围绕青钱柳种质资源筛选、有性繁殖、无性繁殖、定向培育等方面的研究现状进行分析,总结了目前繁育技术研究过程中存在的无性繁殖难生根问题和组织培养过程中的污染、褐化、玻璃化等问题,提出了预期的解决方案和未来的研究方向,旨在为青钱柳进一步的资源培育提供参考。

基于网络药理学探讨青钱柳镇痛作用机制

目的探明青钱柳镇痛作用机制,为青钱柳临床应用和药物开发提供科学指引。方法通过中药网络药理学的研究方法探讨青钱柳镇痛作用机制。利用分子对接验证核心靶点与活性化合物的结合能力。用纯化水对青钱柳叶进行恒温回流提取。用ELISA试剂盒检测青钱柳叶水提物降低小鼠疼痛指标以及对核心靶点的抑制情况。结果青钱柳镇痛的核心靶点为IL-6、COX-2和MMP-9。核心靶点与活性化合物均能较好结合。青钱柳叶水提物能较好地降低小鼠疼痛指标和抑制核心靶点IL-6、COX-2以及MMP-9。结论青钱柳的主要镇痛机制可能通过抑制核心靶点IL-6、COX-2和MMP-9,从而达到镇痛的效果。

不同种源家系青钱柳在福建沙县的生长适应性评价

以南京林业大学种质基因库中优选的16个青钱柳优良种源家系为研究对象,分析该树种在福建沙县地区造林2年内的生长量和主要生物活性物质含量差异,对其适应性进行评价。结果表明:(1)不同种源家系青钱柳在福建沙县地区造林1年和2年后的保存率差异显著。造林1年后,各家系保存率较高,JGS2和JZS24保存率达100%;造林2年后,保存率下降显著,AJ1保存率最高,为80%。JGS2、JZS24和YL1保存率较高且下降幅度小。(2)造林1年和2年后,不同种源家系青钱柳树高和地径年增长值差异不显著。(3)造林2年后,不同种源家系青钱柳主要生物活性物质含量和产量差异显著,其中JZS33表现优良,适应性和生产潜力高,推荐优先选择。JZS4、JZS22、JZS24、JZS27也表现良好,具有推广潜力。通过合理选择和管理优良种源家系,可提高青钱柳造林和生产项目的成功率和经济效益。保存率下降主要因受杉木苗遮阴和高温干旱影响,环境条件对青钱柳生长量和保存率影响显著。

青钱柳叶片甜味性状差异的转录组比较分析

以湖南武冈种植的甜型和非甜型青钱柳叶为材料,分离其RNA并构建cDNA文库,开展2种类型青钱柳的转录组测定。利用转录组数据结合萜类代谢组及其代谢途径分析,研究青钱柳发育过程中甜味性状形成的分子差异。对甜型与非甜型青钱柳叶片转录组的差异基因GO及KEGG分析,共鉴定出13个参与三萜类化合物形成的关键酶基因,在2种青钱柳叶中具有显著差异性表达模式。结合萜类代谢检测结果,甜型青钱柳叶中4类甜味三萜类化合物的积累量显著高于非甜型青钱柳叶的,说明这些基因的差异表达造成了甜味三萜类化合物在叶片中积累量的不同,应是青钱柳甜味差异性状形成的关键原因。

基于VOSviewer和CiteSpace知识图谱的青钱柳可视化分析

目的:通过分析药食同源青钱柳在中医药领域的相关研究文献,了解该药的研究脉络、总结研究热点并预测今后的发展趋势。方法:检索2023年3月1日之前收录在中国知网(CNKI)、维普(VIP)、万方(Wanfang)、中国生物医学文献服务系统(SinoMed)数据库中青钱柳相关文献,使用VOSviewer1.6.19绘制核心作者、机构合作网络,运用CiteSpace6.1.R3进行关键词共现、聚类、突现及时间线分布的可视化分析。结果:最终纳入文献267篇,从发文趋势看,发文量呈逐年增加,发文最多的作者是舒任庚,中文载文量最多的期刊是《中草药》,高频关键词有“化学成分”“多糖”“糖尿病”等。结论:青钱柳治疗糖尿病、高脂血症疗效明确,对糖脂代谢的调控作用是本领域研究热点,探究青钱柳药效物质及其作用机制是本领域的研究趋势。

基于CiteSpace对青钱柳药材研究进展的可视化分析

目的 探讨药材青钱柳药材的研究现状、研究热点、演化路径及发展趋势。方法 采用计算机检索中国知网(CNKI)及Web of Science数据库自建库起至2023年3月1日已发表的青钱柳相关研究的国内外文献,运用文献计量学CiteSpace 6.1. R6软件对研究作者、研究机构、关键词等内容进行知识图谱绘制,并结合信息提取的方法进行可视化分析。结果 最终纳入中文文献709篇,英文文献326篇;年度发文量呈波动上升趋势;方升佐是发表中文和英文文献最多的作者;发文机构分析显示,南京林业大学是发表中文和英文文献最多的研究机构,其中中文文献机构间合作相对零散,英文文献机构间合作相对较多;关键词分析显示,青钱柳研究主要集中在成分分析、药理作用、作用机制、繁殖方法、栽培技术等方面;中文文献聚为25类,英文文献聚为13类;抗氧化、降血糖作用、青钱柳治疗作用及起效机制、青钱柳黄酮、青钱柳对免疫调节的影响等将是医疗健康方面的研究热点,其药用价值越来越受到重视。结论 青钱柳药材的相关研究仍处于发展阶段,需加强作者或机构的跨团体、跨区域合作,加强校-企-政合作,推动产-学-研深度融合。

基于亲和超滤-液质联用结合相关性分析方法筛选青钱柳提取物中α-葡萄糖苷酶抑制剂

目的:为快速筛选青钱柳提取物中的α-葡萄糖苷酶抑制剂,采用亲和超滤-液质联用技术结合相关性分析探究其有效成分与酶抑制活性间的关系。方法:运用大孔吸附树脂,建立青钱柳不同浓度乙醇洗脱液中相同化学成分量变化矩阵(X矩阵);分析各乙醇洗脱部位体外α-葡萄糖苷酶抑制作用,构建效应矩阵(Y矩阵);通过偏最小二乘回归法(Partialleastsquaresregression,PLS)预测潜在的活性成分与亲和超滤法(Affinity ultrafiltration,AUF)所筛选的化合物进行比对并通过分子对接进行验证。结果:青钱柳50%乙醇总提取物IC50值为14.8μg/mL,经大孔树脂吸附后的10%~80%浓度乙醇洗脱液IC50为5.4~204.1μg/mL。通过AUF法与相关性分析在正、负离子模式下共鉴定出10种青钱柳潜在的活性成分,其中鉴定出8种三萜类化合物、2种黄酮类化合物。进一步的分子对接结果表明,上述10种活性成分与α-葡萄糖苷酶对接的结合能小于-7.0 kcal·mol^(-1),其中熊果酸内酯的结合能最低(-10.0 kcal·mol^(-1))。结论:AUF与相关性分析相结合能快速筛选出青钱柳提取物中潜在的α-葡萄苷酶抑制剂,本研究结果可为进一步明确青钱柳活性成分奠定基础,为其活性成分质量控制研究、功能性食品开发的深入研究提供依据。

青钱柳染色体的制片技术及核型分析

采集母树生长于湖北省宜昌市五峰土家族自治县的青钱柳(Cyclocarya paliurus)二倍体和四倍体种子,在室外自然层积至萌发,待根长(L)分别达到0.5 cm<L≤1.0 cm、1.0 cm<L≤2.0 cm、L>2.0 cm时,选取根部嫩白且完整的根尖,按照试验设定的处理条件进行预处理、固定、解离、染色。取长度达到0.5 cm<L≤1.0 cm的二倍体青钱柳胚根的根尖,采用常规压片法,比较预处理条件、解离方法、染色时间对染色体制片效果的影响,获取优化的二倍体青钱柳根尖染色体制片技术;按照优化后的二倍体青钱柳根尖染色体制片技术,对四倍体青钱柳根尖进行染色体制片。对不同倍性青钱柳各选取30个以上染色体分散良好的细胞分裂相进行染色体数目统计,选取5张染色体分散良好、形态清晰的中期细胞照片,采用测量和分析工具(MATO)软件对染色体的长臂、短臂进行测量,计算相对长度和臂比,再进行染色体类型划分、核型分类、核型不对称系数计算。结果表明:①选取根长为0.5 cm<L≤1.0 cm的根尖,4℃避光条件时,用质量浓度为2 g/L的秋水仙素与浓度为2 mmol/L的8-羟基喹啉等体积比例混合溶液处理1~2 h(或对二氯苯饱和水溶液处理6 h)、常温时浓盐酸和乙醇混合溶液解离10 min、卡宝品红溶液染色30 min,制片效果最佳。②二倍体青钱柳核型公式为2n=2x=32=32m,核型不对称系数为57.61%,属于1B型,染色体全由中部着丝粒染色体组成;四倍体青钱柳核型公式为2 n=4x=64=60m+4sm,核型不对称系数为58.83%,属于1B型,染色体由中部和近中部着丝粒染色体组成。③试验建立了青钱柳染色体制片的适宜方案;核型分析表明,青钱柳二倍体和四倍体在进化中均处于较原始地位。

基于临床新用的青钱柳中药药性探讨

青钱柳为双子叶植物纲胡桃科青钱柳属速生乔木,主要分布于我国的南部亚热带的山区,是我国独有的珍贵树种,目前在我国已有大规模种植。青钱柳作为药物最早记载于1975年出版的《全国中草药汇编》,之后又陆续被《中国中药资源志要》《中华本草》等中药专著以及4个地方药材标准收载。但是没有形成统一的药用部位、采收时间、性味归经、功能主治、用法用量的标准,功能主治与当前实际应用存在一定差异,没有紧跟临床进展不断总结并及时更新,尤其缺乏系统的“方-证-症”与之相对应,难以指导青钱柳临床辨证用药和合理用药。青钱柳叶作为别样茶已经有500年以上的悠久历史,于2013年被国家卫生健康委员会列入新资源食品目录。因而青钱柳是一味药食两用中药。以临床应用为依据、实验研究为参考,总结青钱柳现有的研究成果,运用中医药思维对其中药药性进行再探索,完善其中药药性,总结和归纳了功能主治,为统一和提升青钱柳药材标准,为其合理配伍配方和辨证用药提供参考与指导。基于临床新用深入探讨青钱柳中药药性对进一步挖掘其价值,提高使用的准确性、规范性及科学性,更好地为人民群众健康服务具有重要意义。