Anorectal Pharmacodynamics and In Vitro Drug Release of Clerodendrum bungei Steud.Extract Gel

[Objectives]To determine the optimal preparation technology of Clerodendrum bungei Steud.extract gel by orthogonal test and gel quality test method in General Rule 0114 of Chinese Pharmacopoeia(Volume IV,2020 Edition),and to study its anorectal pharmacodynamics and drug release in vitro.[Methods]Carbomer 940,propylene glycol and absolute ethyl alcohol were selected as the main factors,and the preparation technology of C.bungei Steud.extract gel was optimized by orthogonal test.The mouse model of ulcerative hemorrhoids was established with glacial acetic acid(HAC)and compared with Ma Yinglong musk hemorrhoids ointment.The recovery of trauma was compared between the two groups.At the same time,porcine small intestine was used as semi-permeable membrane to make diffusion cell to simulate anal environment,and the drug release in vitro was studied.[Results]The C.bungei Steud.extract gel was smooth in appearance and good in stability.It could effectively treat anal ulcer in mice and release quickly in vitro.[Conclusions]The formula is reasonable,and the effect of animal experiment is remarkable,which can provide a new treatment plan for ulcerative hemorrhoids.

中国楸树(Catalpa bungei C.A.Mey)种质资源遗传多样性的ISSR分析

采用简单序列重复区间标记法(ISSR)分子标记技术,对中国楸树(Catalpa bungeiC.A.Mey)10个类型156个单株进行了遗传多样性分析。从46个简单重复序列引物中筛选出的14个引物共扩增出190条稳定的谱带,其中多态性谱带186条,多态性条带百分率达97.89%。在物种水平上,Nei′s基因多样性指数(H)为0.290 9,Shannon信息指数(I)为0.446 3,基因多样度(Ht)为0.296 7,基因分化系数(Gst)为0.397 1,基因流(Nm)为0.759 2。分析结果表明,楸树各类型间变异占总变异的39.71%,类型内的变异占总变异的60.29%,约为类型间变异的1.5倍,说明中国楸树类型内的变异在其遗传变异中占主导地位。楸树类型间的遗传相似系数介于0.668 4和0.984 9之间,平均为0.863 7。基于遗传相似系数的UPGMA聚类分析,可将楸树10个类型分为4类。

5个楸树(Catalpa bungei)无性系耗水特性比较

水是西北造林育种需要考虑的关键环境因子,速生树种高耗水现象是西北造林中亟待解决的问题。为筛选适宜造林的楸树无性系,本研究利用盆栽称重法及Li-6400XT便携式光合作用分析系统（Li-cor,U.S.）对于5个楸树无性系（004-1、015-1、7080、1-3、1-4）进行比较。结果显示：试验地甘肃天水地区午间14：00时气温达全天最高,为41.1±1.86℃,湿度仅为39.21±2.65%。在此条件下,5个楸树无性系耗水量及耗水速率均出现＂单峰变化＂,可归为＂非蒸腾午休型＂。其中无性系1-3午间耗水速率最大,达到7.9691±0.1367 mmol·m^-2·s^-1;楸树无性系全天耗水量较高,在1630.42±38.27-2089.95±34.14 g·d^-1,平均为1709.30±52.38 g·d^-1,但其日间生产能力较高,日间耗水速率占全天耗水速率92.78%,显著高于毛白杨。与其他4个无性系相比,无性系004-1水分利用效率出现＂双峰＂现象,分别在12：00时及16：00时出现两个峰,全天水分利用效率总和达到15.723±1.231μmol·mmol-1。因此,楸树耗水速率与普通植物相当,耗水量较高的原因与其较大的叶面积有关;无性系004-1较其他4个无性系,具有更高的苗高、地径,且整株叶面积达1.9907±0.032 m^2;在水分利用效率、耗水速率上也表现出良好的水分利用能力和生长能力,可作为当地丰产速生林的优质无性系进行种植。

Protective effects of Radix Cynanchum bungei on ischemia-induced neuronal and cognitive impairment in mice

OBJECTIVE To investigate the effects of Radix Cynanchum bungei extract(RCBE) on cerebral ischemia-reperfusion(I/R) and acute cerebral ischemia induced impairment in mice.METHODS I/R model was induced by bilateral carotid artery occlusion(BCAO)-reperfusion method and Y-maze learning and memory performance was assessed after reperfusion. Na^+-K^+-ATPase,Ca^(2+)-ATPase and SOD activity,as well as MDA content in mouse brain tissue were measured. Numbers of mouth-opening breaths of the isolated mouse head were observed in acute cerebral ischemia mice.RESULTS Learning and memory ability in mice with RCBE were improved significantly compared with model group. The activity of SOD,Na^+-K^+-ATPase and Ca^(2+)-ATPase were increased,while MDA contents decreased after RCBE(0.5,1.0 and 2.0 g·kg^(-1)) and piracetam(0.5 g·kg^(-1)) treatment compared with the model group. RCBE at all concentrations significantly prolonged the number of mouth-opening breaths of the isolated mouse head. CONCLUSION RCBE preconditioning exerts a marked neuroprotective effect on the ischemia brain,which is related to improve the learning and memory via regulating energy metabolism and anti-oxidation.

Physical and Mechanical Properties of Catalpa bungei Clones and Estimation of the Properties by Near-Infrared Spectroscopy

Air-dry density,modulus of rupture(MOR),modulus of elasticity(MOE),compressive strength parallel to grain,and hardness of Catalpa bungei clones were investigated in this study with feasibility of predicting these properties by near infrared(NIR)spectroscopy.The best candidate Luoqiu 3'has been selected from three clones based on wood physical and mechanical property indices.Lower values of wood physical and mechanical properties have been found in the corewood compared to the outerwood.There were significant positive correlations between the air-dry density and mechanical properties.Information from Cross section for air dry density,compressive strength parallel to grain,and hardness yielded prediction models with better effects,along with the best MOR and MOE modeling effects resulted from average sections'spectra collection.Multiplicative scatter correction(MSC)+Savitzky-Golay(S-G)smoothing method has been proved to be the most applicable way.In addition,the predictions from five point sampling method were slightly better than three point one.Overall,results suggest NIR spectroscopy was viable to predict the physical and mechanical properties of C bungei dones with methods developed in this study proved effective in preliminary screening.

Global Lysine Acetylome Analysis of Flower Bud Development in Catalpa bungei

Lysine acetylation,a reversible and dynamic post-translational modification,plays pivotal roles in regulating many biological processes in plants.However,the information of lysine acetylation during flower development of woody angiosperm remains unclear.Herein,we identified a total of 667 lysine acetylation sites in 467 proteins in flower buds of Catalpa Bungei.The acetylated proteins were mainly involved in the pathways of carbon metabolism,glycometabolism and oxidative phosphorylation.Using functional enrichment analysis,61 and 13 acetylated proteins were involved in the glycometabolism and oxidative phosphorylation pathways,respectively.This suggests that lysine acetylated proteins play critical roles in the energy metabolism pathways.Accordingly,the protein–protein interactions were also highly enriched in the carbon metabolic pathway.Integrative analysis of lysine acetylome and transcriptome showed that transcript level changes of seven key lysine-acetylated proteins/genes involved in energy metabolism were significant different during floral transition.Importantly,compared with the normal flowering variety,the expression levels of phosphoribulokinase,phosphoenolpyruvate carboxykinase,ATP synthase and pyruvate dehydrogenase complex genes were significantly up-regulated at the stages of vegetative and transition buds during early flowering of C.bungei.These data represent the first lysine acetylome in the C.bungei flowers,and can serve as an essential basis for exploring the function of lysine acetylated proteins in the flower bud development of plants.

锑胁迫对楸树无性系叶片解剖结构的影响

以10个楸树无性系为研究对象,采用室内盆栽的方法,共设置4种浓度的锑胁迫处理,各处理锑元素含量分别为0、600、1200、2000 mg·kg^(-1);以石蜡切片方法观测叶片的解剖结构,探讨锑胁迫对楸树叶片解剖结构的影响。结果表明:除2-8、1-1号2个楸树无性系在2000 mg·kg^(-1)锑胁迫下其叶片中锑含量均下降外,10个楸树无性系在0~2000 mg·kg^(-1)锑胁迫下其叶片中锑含量均随着锑胁迫浓度的增加而增加。在不同浓度的锑胁迫下,5-8、0号2个无性系的叶片厚度均没有显著差异,其余无性系的叶片厚度表现出不同的差异性;随着锑胁迫浓度的增加,不同楸树无性系的叶片厚度表现出不同的变化规律。随着锑胁迫浓度的增加,10个楸树无性系的叶片上表皮厚度和下表皮厚度总体呈现先升高后下降的变化规律。在不同浓度锑胁迫下,10个楸树无性系的叶片组织结构存在一定差异;在600~2000 mg·kg^(-1)锑胁迫下,各楸树无性系平均栅海比排序为5-8号的(0.99)>1号的(0.91)=2-8号的(0.91)>0号的(0.90)>5-2号的(0.89)>8402号的(0.88)=72号的(0.88)>1-1号的(0.86)=20-01号的(0.86)>63号的(0.71)。0~2000 mg·kg^(-1)锑胁迫对楸树无性系叶片结构无明显破坏,楸树对锑具有一定的耐受性。

楸树DELLA基因家族生信分析及CbuGRAS9的功能分析

通过鉴定楸树(Catalpa bungei)DELLA家族基因并分析CbuGRAS9的基因功能,为楸树生殖调控性状的遗传改良提供理论依据。基于楸树基因组数据,鉴定并克隆了5个与拟南芥(Arabidopsis thaliana)同源的CbuDELLAs基因;利用ExPASy、SWISS-MODEL、Plant-mPloc、PlantCare等在线工具对CbuDELLAs蛋白进行等电点、蛋白结构、亚细胞定位及启动子顺式作用元件预测;以9-1(Catalpa bungei‘Luoqiu No.1’)和‘百日花’楸树(Catalpa‘Bairihua’)为材料,分析了CbuDELLAs基因的表达量差异,并通过异源转化拟南芥证实了CbuGRAS9的分子功能,利用酵母双杂交文库筛选了与CbuGRAS9互作的蛋白。结果表明:5个CbuDELLAs蛋白的氨基酸数目为455~588,蛋白相对分子质量为5.04~6.43 kDa,等电点为4.81~5.14;CbuDELLAs蛋白均含有DELLA和GRAS保守结构域,全部为亲水性蛋白。亚细胞定位预测结果显示CbuDELLAs蛋白均定位于细胞核中。启动子顺式作用元件分析发现,这5个DELLAs基因启动子区均含有参与赤霉素反应的顺式作用元件。qRT-PCR结果显示‘,百日花’楸中CbuDELLAs基因表达量显著高于对照9-1楸,其中CbuGRAS9为差异最显著的基因,CbuGRAS9转基因株系的开花时间被明显推迟。鉴定到与CbuGRAS9互作的蛋白质主要富集在核糖体、氨基酸合成、次级代谢、光合作用、TCA循环等代谢通路。

楸树远缘杂交种花器官性状变异分析

为明确楸树(Catalpa bungei)远缘杂交种花器官性状的表型变异程度和变异规律,以12株楸树远缘杂交种为材料,采用方差分析、相关分析、聚类分析等统计分析方法,对楸树远缘杂交种的花期物候特征、花序性状、花朵性状和花色性状指标进行研究。结果表明:楸树远缘杂交种的花期物候特征差异明显,楸梓杂交种为假三叉分枝,花芽分化能力强,嫁接当年持续多轮开花,花期长达100 d以上,表现出母本性状。楸梓杂交种的花器官特征差异显著,花序轴共7~9轮侧枝,主轴为无限花序,侧枝为二歧聚伞花序,从底层到顶层过度为聚伞花序,处于楸树有限花序到梓树无限花序的过渡性状。花总长度、花筒直径、花筒长度、花冠宽度、花梗长度、雌蕊长度、花药长度、雄蕊长度、不育雄蕊长度的变幅分别为35.42~50.36,10.84~16.13,16.57~24.68,31.58~44.87,9.07~31.99,19.44~27.52,4.85~5.89,13.93~21.10,3.66~6.79 mm,变异系数为10.88%~31.02%,多样性指数(H’)为1.36~1.93,并以9个性状指标将12株杂交种聚为大花型、中花型、小花型3类。楸树杂交种间花色差异显著,以L*、a*、b*值将12株杂交种的唇瓣花斑颜色聚为3类,分别是粉紫色型、紫红色型和紫色型。

长期继代培养过程中楸树愈伤组织的分化能力

以长期继代培养的楸树愈伤组织和新诱导的楸树愈伤组织为试验材料,利用石蜡切片比较分析它们的组织形态特征,并测定过氧化物酶(POD)、超氧化物歧化酶(SOD)和淀粉酶的活性,以及可溶性蛋白与可溶性糖的含量,以探究楸树愈伤组织长期继代培养后分化再生困难的形成原因。组织形态学观察分析发现,继代培养7 a的白色颗粒状愈伤组织细胞中淀粉粒含量丰富,分布均匀,仍为胚性愈伤组织;新诱导的浅绿色愈伤组织存在明显的细胞分化中心,有均匀分布的淀粉粒,为早期胚性愈伤组织;新诱导的黄白色致密愈伤组织为非胚性向胚性转化的愈伤组织,新诱导黄白色疏松愈伤组织为非胚性愈伤组织。生理生化指标测定分析发现,继代培养7 a的楸树胚性愈伤组织中可溶性糖和可溶性蛋白的含量均显著高于新诱导出的3种愈伤组织,但其同工酶活性却较低,表明在长期继代培养的过程中,楸树愈伤组织的生理生化代谢活性降低。由此推测生理生化代谢活性的下降可能是造成楸树胚性愈伤组织长期继代培养后分化再生困难的重要因素。

金丝楸抑菌皂制作及活性评价

采用浸提和柱层析等手段从金丝楸心材中制备抗菌组合物,该组合物体外抗金黄色葡萄球菌(S.aureus ATCC 25923)最小抑菌浓度(MIC)值为64.0μg/mL。运用HPLC技术,结合标准品比对法,鉴定该组合物主要化学成分为梓木内酯和4,9-二羟基-α-拉帕醌,相对含量,分别为62%和23%。通过添加一定量的金丝楸抗菌组合物,制得金丝楸抑菌皂,并评价了药物浓度与抑菌活性的相关性。当添加浓度为90μg/mL时,制得的抑菌皂与市售三种抑菌皂抑菌效果一致。

The Total Flavonoids of Clerodendrum bungei Suppress A549 Cells Proliferation,Migration,and Invasion by Impacting Wnt/β-Catenin Signaling

Objectives: The objective of this study is to evaluate the effect of the total flavonoids of Clerodendrum bungei(TFCB) on the proliferation,invasion, and metastasis of A549 lung cancer cells through the Wnt signaling pathway. Materials and Methods: A549 cells were transfected with a β-catenin overexpression plasmid and the empty vector pcDNA3.1. The A549 cells were divided into six groups: normal A549 cell group, normal A549 cells with TFCB group, vector control group, vector with TFCB group,(3-catenin overexpression group, and β-catenin with TFCB group. We used the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to detect cell proliferation, a scratch test was used to observe cell migration, and a transwell experiment was employed to evaluate cell invasion. Proteins related to the Wnt pathway were detected with Western blot analysis, including β-catenin, GSK-3 β, P-GSK-3 β, c-Myc, and CyclinD1. Results: The proliferation, invasion,and metastasis of A549 cells were significantly enhanced after being transfected with the β-catenin overexpression plasmid(P< 0.05 or 0.01),accompanied by increased expression of β-catenin, C-Myc, CyclinD1 and reduced expression of Gsk-3 β and P-GSK-3 β. Treatment of cells with TFCB resulted in inhibition of cell proliferation, migration, and invasion; downregulated expression of β-catenin, C-Myc, and CyclinD1;and upregulated expression of GSK-3 β and P-GSK-3 β,especially in the β-catenin overexpression group. Conclusion: TFCB has the potential to inhibit the Wnt/β-catenin pathway by prohibiting the overexpression of β-catenin and regulating its downstream factors.

浅析大规格楸树圃地苗抚育管理技术——以界首市为例

基于楸树生物学特性,以界首市为例,阐述了大规格楸树圃地苗的抚育管理技术和要点。通过做好松土除草和水肥管理工作、科学定植与施肥、针对性地开展病虫害防治,为培育优质大规格楸树壮苗提供科学依据。

不同楸树品种根段及枝干萌条内源激素质量分数及扦插生根率

以3个楸树品种的1年生根段萌条和枝干萌条为材料,对其进行内源激素质量分数测定和扦插生根率统计。结果表明:楸树3个品种根段萌条的生根率皆高于枝干萌条,且不同品种间,根段萌条和枝干萌条的生根率都存在极显著差异,3个品种的生根率从大到小依次为楚楸1号、洛楸1号、金丝楸;除了洛楸1号的茉莉酸和金丝楸的DL-二氢玉米素、赤霉素3的质量分数在根段萌条低于其枝干萌条外,其余激素在不同品种根段萌条中的质量分数皆高于对应枝干萌条;不同部位萌条内源激素质量分数的差异随品种不同而不同,洛楸1号有4种激素在两个不同部位萌条中存在显著差异,楚楸1号有1种存在显著差异,而金丝楸中所有激素都差异不显著;不同品种相同部位萌条中内源激素间差异分析结果表明,除脱落酸外,根段萌条中其他内源激素质量分数间的差异均大于枝干萌条;根段萌条各因素间整体相关性及显著性强于枝干萌条各因素间,反式玉米素、DL-二氢玉米素与生根率存在显著相关性。

臭牡丹苯乙醇苷类成分的鉴定及体外抗人肺腺癌A549细胞的机制研究

臭牡丹提取物具有良好的抗肿瘤的作用,为了鉴定臭牡丹提取物的成分及探究其对A549细胞体外抑制的影响,采用超高效液相色谱-质谱联用技术、CCK-8、细胞划痕法、Transwell、Western blot、免疫荧光及流式细胞术等方法进行检测。鉴定了臭牡丹提取物中34个苯乙醇苷类化合物,其中类叶升麻苷含量为32.95%,异类叶升麻苷5.49%。体外实验表明,臭牡丹提取物能抑制A549细胞的增殖、迁移和侵袭,且IC 50为0.125 mg/mL,还能下调A549细胞中EphA2、p-AKT、mTOR、p-mTOR、p-GSK-3β和β-catenin的表达,上调EphrinA1的表达,减少EphA2在A549细胞膜与膜质上的表达,并将A549细胞周期阻滞于G1/S期。本研究证实臭牡丹地上部分富含苯乙醇苷类成分,尤以类叶升麻苷和异类叶升麻苷为多,且可通过调控EphA2/Akt/mTOR通路达到抗肺癌作用。

恩施楸树半同胞家系幼林综合评价

为了比较恩施楸树半同胞家系幼林表现差异,以恩施地区收集的天然楸树种子所育实生苗为材料造林,通过测量不同半同胞家系的生长、叶绿素、光合特征等,利用模糊函数对不同半同胞家系进行综合评价。结果显示,不同家系间胸径差异没有达到显著水平(P>0.05),而树高差异则达到了显著水平(P<0.05),这主要与栽植密度过大有关,不同家系间叶片叶绿素含量差异达到了极显著水平(P<0.01)。不同家系表现出不同的光强适应性,‘7号’和‘54号’家系光饱和点均超过1700μmol/(m^(2)·s),对强光有着较好的适应性,‘8号’家系最大净光合速率达到23.24μmol/(m^(2·)s),具有较大的光合潜力,‘11号’和‘8号’家系有较好的弱光适应性,‘7号’和‘8号’家系暗呼吸速率相对较高,代谢活跃。树高、叶片叶绿素含量、光饱和点、最大净光合速率、光补偿点、暗呼吸速率等均与胸径呈正相关,几个家系的综合评价排序为‘8号’>‘54号’>‘9号’>‘7号’>‘12号’>‘11号’‘,8号’家系选育潜力巨大。

金丝楸幼苗响应盐碱胁迫的生理和转录组分析

[目的]研究不同盐碱胁迫对金丝楸幼苗生长、光合和生理指标的影响,并结合转录组测序分析,探究楸树耐盐碱的生理机制和分子机制。[方法]采用盆栽法对金丝楸幼苗进行不同盐碱胁迫处理,分析其生物量、光合及生理指标对不同盐碱响应的差异,采用Illumina高通量测序技术进行转录组测序,通过生物信息学分析盐碱胁迫对转录水平的影响。[结果]不同盐碱胁迫下,金丝楸幼苗叶片受伤害程度为Na_(2)CO_(3)>混合盐碱>NaCl;新增株高和地径、地上部和根的干质量和鲜质量、生物量、根冠比均受到明显抑制,并随盐碱浓度增加而抑制加强,但生长胁迫指数均随浓度的增加而降低;丙二醛(MDA)含量和相对电导率都随胁迫浓度增加而不同程度上升,超氧化物歧化酶(SOD)活性、可溶性糖含量、脯氨酸(Pro)含量、叶绿素总量和光合速率都呈现先上升后下降的趋势。转录组测序共产生约60.4 Gb原始数据,组装得到55793个Unigenes,其中29534(52.93%)个Unigenes获得了注释;通过差异表达基因(DEGs)分析,3个比较组(CK vs NaCl,CK vs Na_(2)CO_(3)和CK vs混合盐碱)分别筛选出1779、2835和4059个DEGs;DEGs GO富集分析表明,膜的整体成分、膜的内在成分、催化活性、类异戊二烯代谢和合成过程、氧化还原酶活性等条目被显著富集;DEGs KEGG分析表明,苯丙素生物合成、淀粉和蔗糖代谢、植物激素信号转导、萜类主干生物合成和精氨酸代谢等通路被显著富集;此外,在DEGs中鉴定的bHLH、ERF、MYB-related、NAC、C2H2、WRKY、MYB和b ZIP转录因子家族成员最多。[结论]金丝楸主要通过积累可溶性糖和Pro,提高SOD酶活和光合作用来抵御盐碱胁迫,但都呈现“低促高抑”的现象,说明其具有一定阈值。金丝楸通过调节膜成分、催化活性、类异戊二烯代谢和生物合成过程、苯丙素生物合成、淀粉和蔗糖代谢、植物激素信号转导等生物过程和代谢途径,并结合有关转录因子共同响应盐碱胁迫。本研究为深入研究楸树耐盐碱生理机制和分子机制提供科学的理论依据。

利用近红外光谱技术快速预测楸树木材抗弯性质

楸树(Catalpa bungei)木材纹理通直、材性优良、用途广泛,是中国特有的珍贵材树种。研究木材重要的力学性质——抗弯性质的快速测定方法可以为楸树木材的遗传改良及加工利用提供科学依据。以楸树无性系新品种“洛楸1号”、“洛楸4号”和“天楸2号”为试验材料,依据国家标准抗弯性质测试方法,测定楸树木材抗弯强度(MOR)和抗弯弹性模量(MOE)。利用近红外光谱(NIRs)分析结合偏最小二乘法(PLS)对新选育的三个楸树无性系的抗弯性质进行预测,探究基于不同采谱切面、不同预处理方法以及不同采谱点的最佳建模方法。研究结果表明,基于两切面平均光谱建立的抗弯强度预测模型的相关系数和相对分析误差最高为0.843和1.88,建立的抗弯弹性模量预测模型的相关系数和相对分析误差最高为0.846和1.88。选用两切面光谱,预处理方法按抗弯强度模型性能排序为多元散射校正与卷积平滑结合算法(MSC+S-G)>二阶导数与卷积平滑结合算法(2^(nd)Der+S-G)>一阶导数与卷积平滑结合算法(1^(st)Der+S-G),预处理方法按抗弯弹性模量模型性能排序为MSC+S-G>1^(st)Der+S-G>2^(nd)Der+S-G。使用一点采谱法建立的抗弯强度和抗弯弹性模量预测模型相关系数比五点采谱法的分别降低5.93%和2.96%。综上所述,近红外光谱可以用于预测珍贵材楸木的抗弯强度和抗弯弹性模量。采用不同切面、预处理方法和采谱点数建立的模型,建模结果有一定差异。得出了楸树木材抗弯强度和抗弯弹性模量的最佳建模方法。基于径切面和弦切面平均光谱建立的抗弯强度和抗弯弹性模量近红外模型效果最佳。MSC+S-G是最适用于楸树木材抗弯性质的预处理方法。五点采谱法模型精度较高,但对大量样品抗弯性质快速估算时,可以降低采谱点数量,仅采集中间部位即载荷加载部位一个光谱点以减少采谱工作量,提高楸树木材抗弯性质快速评估效率。

臭牡丹调控脊髓N-甲基-D-天冬氨酸受体亚基缓解神经病理性疼痛

目的:探讨臭牡丹(Clerodendrum bungei Steud.,CBS)的镇痛作用是否与N-甲基-D-天冬氨酸(N-methyl-Daspartate,NMDA)受体相关。方法:雄性成年SD大鼠24只,随机分为假手术组(Sham组)、坐骨神经分支选择性损伤(Spared nerve injury,SNI)模型组(SNI组)和30 g·kg^(-1)CBS治疗组(SNI+CBS组)。SNI+CBS组于SNI术后第7 d灌胃给予CBS,持续21 d,Sham组和SNI组给予同等体积的生理盐水灌胃。于术后第26~28 d检测大鼠步态;第28 d取脊髓,qPCR方法检测NMDA受体亚基GluNR1、GluN2A、GluN2B和GluN3A mRNA表达,Western blot和免疫荧光方法检测GluN2A和GluN2B蛋白水平,分子对接方法模拟CBS中主要成分与GluN2B的亲和力。结果:与Sham组相比,SNI模型组大鼠患肢的站立时间、最大接触时的最大强度、足印长度、足印宽度和爪印面积降低,最大接触时间百分比升高,脊髓GluN2A、GluN2B和GluN3A mRNA表达降低,GluN2A和GluN2B蛋白水平升高。与SNI组相比,SNI+CBS组大鼠患肢的站立时间、最大接触时的最大强度、足印长度和足印宽度升高,最大接触时间百分比降低,脊髓GluN2A、GluN2B和GluN3A mRNA表达升高,GluN2A和GluN2B蛋白水平降低。各组GluNR1 mRNA表达差异无统计学意义(P>0.05)。分子对接实验结果显示,原始配体Ro25-6981与GluN2B对接的结合自由能(△G)为−11.72 KJ·mol^(-1),估计抑制常数(Ki)为2.58 nmol;CBS主要成分α-香树脂醇、赪桐酮和木栓酮与GluN2B对接的△G及Ki与Ro25-6981接近,甚至优于Ro25-6981;从对接构象上看,Ro25-6981与木栓酮占据了GluN1B结合位点,α-香树脂醇和赪桐酮则占据了GluN2B结合位点,呈现出较强的NMDA受体抑制作用。结论:CBS对SNI模型所致神经病理性疼痛具有缓解作用,其机制可能是通过抑制NMDA受体来实现。

两种砧木楸树嫁接苗生长差异及转录组比较分析

嫁接是楸树(Catalpa bungei)良种扩繁的主要途径。为探讨以梓树和滇楸为砧木的两种‘苏楸1号’良种当年生嫁接苗生长差异,比较了二者的嫁接成活率、新梢长度、新梢粗度、接穗粗度/砧木粗度、叶面积等,以筛选较佳砧木,并通过高通量测序平台对两种嫁接苗叶片和顶芽进行转录组测序分析,采用实时荧光定量PCR技术对初步筛选的差异表达基因进行验证。结果显示,以梓树为砧木的嫁接成活率和嫁接苗各生长量均显著高于以滇楸为砧木的嫁接苗;转录组测序筛选出两个组合叶片和顶芽差异表达基因共计559个,其中上调表达基因192个,下调表达基因367个;GO富集分析表明559个差异表达基因显著富集在代谢过程、细胞过程、细胞器、结合和催化活性等功能类别;KEGG富集分析表明叶片中213个差异表达基因显著富集到66条代谢途径,顶芽中137个差异表达基因显著富集到45条代谢途径;初步筛选出的Unigene0040270、Unigene0006320、Unigene0036149和Unigene0007805等4个差异表达基因的RT-qPCR趋势与转录组数据一致。本研究结果发现两种嫁接苗的生长存在显著差异,其中以梓树为砧木的嫁接苗各生长参数更好,同时在转录组角度上对其差异进行了初步解析,为楸树嫁接分子研究提供一定的参考价值。