Transcriptome-Wide Identification and Functional Analysis of PgSQE08-01 Gene in Ginsenoside Biosynthesis in Panax ginseng C.A.Mey.

Panax ginseng C.A.Mey.is an important plant species used in traditional Chinese medicine,whose primary active ingredient is a ginsenoside.Ginsenoside biosynthesis is not only regulated by transcription factors but also controlled by a variety of structural genes.Nonetheless,the molecular mechanism underlying ginsenoside biosynthesis has always been a topic in the discussion of ginseng secondary metabolites.Squalene epoxidase(SQE)is a key enzyme in the mevalonic acid pathway,which affects the biosynthesis of secondary metabolites such as terpenoid.Using ginseng transcriptome,expression,and ginsenoside content databases,this study employed bioinformatic methods to systematically analyze the genes encoding SQE in ginseng.We first selected six PgSQE candidates that were closely involved in ginsenoside biosynthesis and then identified PgSQE08-01 to be highly associated with ginsenoside biosynthesis.Next,we constructed the overexpression vector pCAMBIA3301-PgSQE08-01 and the RNAi vector pART27-PgSQE08-01 and transformed ginseng adventitious roots using Agrobacterium rhizogenes,to obtain positive hairy-root clones.Thereafter,quantitative reverse transcriptionpolymerase chain reaction and high-performance liquid chromatography were used to determine the expression of relevant genes and ginsenoside content,respectively.Then,we focused on the function of PgSQE08-01 gene,which was noted to be involved in ginsenoside biosynthesis.Thus,these findings not only provided a molecular basis for the identification of important functional genes in ginseng but also enriched genetic resources for the biosynthesis of ginsenosides using synthetic biology.

High-throughput transcriptional profiling of perturbations by Panax ginseng saponins and Panax notoginseng saponins using TCM-seq

Panax ginseng(PG)and Panax notoginseng(PN)are highly valuable Chinese medicines(CM).Although both CMs have similar active constituents,their clinical applications are clearly different.Over the past decade,RNA sequencing(RNA-seq)analysis has been employed to investigate the molecular mechanisms of extracts or monomers.However,owing to the limited number of samples in standard RNA-seq,few studies have systematically compared the effects of PG and PN spanning multiple conditions at the transcriptomic level.Here,we developed an approach that simultaneously profiles transcriptome changes for multiplexed samples using RNA-seq(TCM-seq),a high-throughput,low-cost workflow to molecularly evaluate CM perturbations.A species-mixing experiment was conducted to illustrate the accuracy of sample multiplexing in TCM-seq.Transcriptomes from repeated samples were used to verify the robustness of TCM-seq.We then focused on the primary active components,Panax notoginseng saponins(PNS)and Panax ginseng saponins(PGS)extracted from PN and PG,respectively.We also characterized the transcriptome changes of 10 cell lines,treated with four different doses of PNS and PGS,using TCM-seq to compare the differences in their perturbing effects on genes,functional pathways,gene modules,and molecular networks.The results of transcriptional data analysis showed that the transcriptional patterns of various cell lines were significantly distinct.PGS exhibited a stronger regulatory effect on genes involved in cardiovascular disease,whereas PNS resulted in a greater coagulation effect on vascular endothelial cells.This study proposes a paradigm to comprehensively explore the differences in mechanisms of action between CMs based on transcriptome readouts.

Based on network pharmacology and molecular docking technology to explore the mechanism of Panax notoginseng in the treatment of membranous nephropathy

Objective:Based on network pharmacology and molecular docking technology to explore the mechanism of Professor Cao Enze's application of Panax notoginseng in the treatment of membranous nephropathy.Methods:TCMSP database was used to obtain the effective components and corresponding target information of Panax notoginseng,and Gene Cards database was used to obtain the disease target genes of membranous nephropathy.The intersection targets of the two were taken and the Venn diagram was drawn.The STRING database was used to obtain the protein interaction relationship,and the PPI network diagram was constructed by Cytoscape 3.9.1 software to screen out the core targets of Panax notoginseng in the treatment of membranous nephropathy.GO function and KEGG pathway enrichment analysis were performed using the David database to obtain the potential pathway of Panax notoginseng in the treatment of membranous nephropathy.Finally,Autodock software was used to verify the molecular docking of the main active components of the drug with the core targets.Results:A total of 7 effective components such as quercetin,ginsenoside rh2,Mandenol and Stigmasterol were retrieved,and 127 potential targets of Panax notoginseng in the treatment of membranous nephropathy were screened out.By PPI network topology analysis,23 core targets such as JUN,TP53,RELA,AKT1 and MAPK1 were screened out.GO functional enrichment analysis contained 703 biological processes,55 cell components and 121 molecular functions,and KEGG signal pathway enrichment analysis enriched 171 signal pathways.The results of molecular docking showed that there was a strong binding ability between the main core targets and the main components of Panax notoginseng.Conclusion:Through network pharmacology,it is concluded that Panax notoginseng treats membranous nephropathy through multiple targets and multiple pathways,which provides a theoretical basis for subsequent basic research.

Panax属西洋参、人参及三七的蛋白电泳指纹鉴别

目的:建立Panax属西洋参、人参及三七的蛋白电泳指纹鉴别法。方法:Western blot及Tris-tricine电泳辅助SDS-PAGE,并测定各区带分子量。结果:经标准化处理的Panax属蛋白电泳图分为三个特征指纹区,由28KD至58KD及其中55KD等2～3条多肽组成三种药材共有蛋白指纹区,对该属鉴别意义重大。28KD以下指纹区西洋参由4～5条区带组成,这是该品种独特的指纹图,58KD以上指纹区对人参鉴别有意义。结论:蛋白电泳指纹图可用于Pnax属药材,特别是西洋参的专属性鉴别。

不同中药材对三七黑斑病病原Alternaria panax Whetz抑制作用研究

通过不同中药材对黑斑病病原Alternaria panax Whetz.室内抑菌试验研究及病原田间致病性鉴定,筛选出可与三七混种的中药材品种泽兰、山萘、麦冬、玉竹。

Anti-tumor Activity of Derivatives of Protopanaxadiol Prepared with Acid Anhydrides

[Objectives]This paper aimed to prepare derivatives of protopanaxadiol from Panax notoginseng(Burk.)FH Chen with acid anhydrides and study their anti-tumor activity.[Methods]The 3-hydroxyl group of protopanaxadiol was subjected to structural modification and reacted with acid anhydrides to prepare derivatives,in order to improve the anti-tumor activity of protopanaxadiol.None of the five compounds designed and synthesized had been reported in the literature,and they were novel compounds.The anti-tumor activity of the derivatives was studies using MTS method.Taking cisplatin and paclitaxel as positive control drugs,the bioactivity of the compounds 1-5 on anti-tumor cell lines(HL-60 cells,SMMC-7721 cells,A-549 cells,MCF-7 cells and SW480 cells)in vitro was screened.[Results]The compound 5 showed inhibitory effect on HL-60 cells,SMMC-7721 cells and A-549 cells.[Conclusions]The acid anhydride esterification method is simple to operate and easy to control.This study has reference value for the structural modification and anti-tumor activity research of protopanaxadiol from P.notoginseng(Burk.)FH Chen.

Minor Saponins from the Leaves of Panax ginseng C.A. Meyer

Five minor compounds isolated from the leaves of Panax ginseng C. A. Meyer were characterized as 20（R）-protopanaxatriol （1）, daucosterin （2）, 3β, 12β-dihydroxy-dammar-20 （22）, 24-diene-3-O-β-D-glucopyranoside （3）, 20 （R）-protopanaxadiol-3-O-β-D-glucopyranoside （4） and ginsenoside-Rh2 （5）, respectively, on the basis of spectral analyses and chemical evidence. The two new saponins, 3 and 4, were named as ginsenoside-Rh3 and 20（R）-ginsenoside-Rh2.Nine other major saponins obtained simultaneously were identical with ginsenoside-Rh1（6）,-Rg3 （7）, -Rg2 （8）, -Rg1 （9）,-Re（10）,-Rd （11）, -Rc （12）, -Rb2（13） and Rb1 （14）, respectively.

人参(Panax ginseng)根系分泌物成分对人参致病菌的化感效应

采用室内培养结合生物学测定的试验方法,研究了不同浓度人参(Panax ginseng)根系分泌物成分苯甲酸、邻苯二甲酸二异丁酯、十六酸和2,2二-(4羟-苯基)丙烷对人参立枯丝核菌(Rhizoctonia solani)、黑斑菌(Alternaria panax)、疫病菌(Phytophthora cactorum)、菌核菌(Sclerotinia schinseng)、锈腐菌(Cylindrocarpon destructans)和绿色木霉菌(Trichoderma viride)菌落生长及孢子萌发的化感效应。结果显示,不同浓度人参根系分泌物成分对人参致病菌及绿色木霉菌的化感效应存在显著差异。苯甲酸浓度与人参立枯丝核菌、菌核菌和锈腐菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关,与人参黑斑菌、绿色木霉菌菌落生长呈正相关;对人参疫病菌菌落生长的化感效应表现为低浓度和高浓度抑制,中浓度促进。邻苯二甲酸二异丁酯浓度与人参立枯丝核菌、黑斑菌、菌核菌和绿色木霉菌菌落生长以及人参黑斑菌孢子萌发呈负相关;对人参锈腐菌菌落生长和孢子萌发表现为低浓度和高浓度抑制,中浓度促进;对人参疫病菌菌落生长表现为低浓度和中浓度抑制,高浓度促进。2,2二-(4羟-苯基)丙烷浓度与人参立枯丝核菌、黑斑菌、疫病菌、绿色木霉菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关;对人参菌核菌、锈腐菌菌落生长表现中浓度促进,高浓度抑制。十六酸浓度与人参锈腐菌、疫病菌和绿色木霉菌菌落生长呈正相关,与人参锈腐菌孢子萌发呈负相关,对黑斑菌孢子萌发表现为中浓度抑制。4种根系分泌物的等量混合物浓度与人参致病菌及拮抗木霉菌菌落生长速率呈负相关。

文山三七(Panax notoginseng)种植区三七与土壤中Pb、Cd、Cu和Zn的分布特征及评价

为了调查文山州三七(Panax notoginseng)主要种植区(丘北县、砚山县、文山县和广南县)土壤和三七Pb、Cd、Cu和Zn含量,通过GPS定位采集三七根区0~15 cm深度土壤样品和三七样品各30个,分析土壤中重金属总量和不同形态含量及三七中对应重金属含量,并进行空间分布分析。结果表明:(1)土壤Pb、Cd、Cu和Zn平均含量分别为55.56、0.36、43.53和119.62 mg·kg^(-1),超标率分别为6.67%、53.3%、13.33%和0。各形态Pb、Cu和Zn含量表现为残渣态>有机物结合态>铁锰氧化物结合态>碳酸盐结合态>可交换态。各形态Cd含量表现为铁锰氧化物结合态>残渣态>碳酸盐结合态>有机物结合态>可交换态。(2)三七和土壤Pb、Cd、Cu和Zn总量和不同形态含量均以丘北县最高,广南县最低。(3)三七根系Pb、Cd、Cu和Zn平均含量分别为2.93、0.35、5.21和11.11mg·kg^(-1),三七植株各部位重金属含量表现为根系>剪口>茎叶>花果。调查区三七和土壤重金属含量存在空间差异,且以土壤Cd污染为主,应采取一定措施降低三七Cd含量。

三七(Panax notoginseng)根残体化感自毒效应研究

为研究三七(Panax notoginseng(Burk)F.H.Chen)采收过程中残留须根的化感自毒效应,采用土培和水培2种方法,按不同比例添加三七须根粉碎物,检测根残体作用下土培三七土壤中皂苷的动态变化及水培三七根部形态结构变化。结果显示,土培条件下,随着须根粉碎物处理时间的延长,土壤中皂苷成分种类增加、总皂苷含量减少。水培条件下,随着处理时间的延长,添加须根粉碎物处理后的三七根尖细胞壁增厚,细胞中出现菌丝体;随着处理时间及处理浓度的增加,细胞开始皱缩甚至破碎,细胞中无完整细胞器结构,三七根部细胞结构差异明显。研究表明三七采收过程中残留在土壤中的须根腐解释放的化学物质可能是导致三七自毒效应的因素之一。

生物质炭、有机肥和钙镁磷肥对三七(Panax Notoginseng)Cd含量的影响

选择生物质炭、钙镁磷肥、有机肥三种改良剂,在云南三七主产区进行田间试验,比较不同改良剂对降低五加科人参属三七（Panax Notoginseng）Cd含量的效果。结果表明,生物质炭和钙镁磷肥处理均显著降低了三七主根、剪口、茎、叶的Cd含量,降低幅度分别为25.4%~43.6%、40.2%~40.9%、34.3%~51.2%和33.0%~33.5%,且生物质炭、钙镁磷肥处理下三七主根干重较对照分别显著提高48.7%和50.4%;生物质炭和钙镁磷肥处理土壤有效Cd含量分别减少56.1%和58.1%,表明生物质炭和钙镁磷肥能有效降低土壤Cd生物有效性、抑制三七Cd吸收。这与生物质炭和钙镁磷肥处理通过降低土壤酸性、提高土壤CEC及有机质含量有关。有机肥处理三七植株生物量和三七各部位Cd含量与对照相比均无显著差异。此外,生物质炭和钙镁磷肥处理显著降低了三七主根、剪口、茎、叶Cd的富集系数（Accumulation coefficient,AF）,对三七Cd转移系数（Transfer coefficient,TF）影响则不显著,而有机肥处理对三七Cd的AF与TF均无影响;各处理三七须根Cd的AF在2.84~4.64之间,显著高于其他部位,而三七主根、剪口、茎、叶等部位Cd的AF和TF均小于1,表明三七须根对土壤Cd富集能力较强而转移能力较差,Cd易集中于三七地下部,Cd污染土壤中施用生物质炭与钙镁磷肥能有效降低Cd在三七体内的富集。

文山三七(Panax notoginseng)种植区土壤As空间分布特征及理化性质对三七As含量的影响

三七（Panax notoginseng（Burk.）F.H.Chen）是云南文山州的道地中药材,文山州由于地壳原始分化成分的作用和含砷矿的开采等原因,使三七种植区存在普遍的土壤砷污染现象。通过野外大田调查和室内分析相结合的方法,研究了云南文山三七主要种植区（文山县、丘北县、砚山县、广南县）土壤总砷和有效砷的空间分布特征和土壤物理化学特征（pH值、有机质含量、阳离子交换量CEC、总P、速效P、土壤质地）,探讨了三七不同部位（茎叶、花果、主根、须根、剪口）As含量的影响因素。将有助于了解文山三七种植区砷污染空间分布特点,寻求三七砷含量的调控途径,对降低三七砷含量和提高三七品质具有一定的实践和理论意义。研究结果表明,（1）土壤有效As和总As平均质量分数分别为0.29 mg·kg-1和28.51mg·kg-1,土壤有效As占总As质量分数的0.06%~2.08%。76.67%的土壤样品总As含量达到国家土壤环境质量的二级标准。土壤有效As和总As含量空间分布表现出西北高,东南低的趋势。（2）三七各部位中As质量分数为0.14~0.32 mg·kg-1,砷含量符合优质三七砷的限量标准（≤1.0 mg·kg-1）,三七各部位对As的富集系数为0.004~0.011,均较低。（3）90%的土壤为酸性土壤,pH值≤6.5;土壤全P质量分数为1.27 g·kg-1,土壤速效P质量分数为66.87 mg·kg-1,均属于丰富水平;土壤有机质质量分数为15.77 g·kg-1,CEC为26.74 cmol·kg-1,属于较高水平;40%的土壤为黏土,43.4%为壤质黏土。（4）土壤总As和土壤有效As含量主要受到随机人为因素的影响,表现出与土壤CEC、pH值和有机质含量之间显著的正相关关系。三七须根中As含量与土壤总As、有效As、土壤黏粒含量、CEC、pH值和总P含量之间具有显著的正相关性。总之,壤质黏土、低pH值和CEC有利于降低三七As含量,三七种植过程中应避免施用含As有机肥和磷肥。研究结果在系统了解文山三七种植区的土壤As空间分布和调控途径方面具有一定的创新价值和意义,特别需要关注是三七产业的发展布局中,应该进行适当的引导和科学的安排三七种植区。

人参(Panax ginseng)的氮过剩现象──人参氮毒原因初探

1985-1990年间，我们在长白县及公主岭进行的四次NPK三要素试验中，发现氮肥抑制了人参生长发育，降低了产量（5％显著水准），而倍量氮肥，减产达1％显著水平。继续研究证明，人参硝酸还原酶活力（NRA）甚低，追施氮肥使组织，积累，促进了呼吸作用，植株干物中N、Si及Cu、Zn、Fe、Mn含量增加，Ca、Mg、K、Na等阳离子减少，表现出氮毒征象。本文称氮过剩症，它是指氮素供应超过了由极低NRA催化的转化代谢速率，而引起的一种毒害现象。

连续2年土壤砷胁迫对三七(Panax notoginseng)细胞膜透性和抗氧化酶活性的影响

通过盆栽试验和室内分析相结合的方法,研究不同砷处理(As5+:0、20、80、140、200和260 mg/kg,采用Na3As O4配制)对三七营养生长期叶片中的细胞膜透性、丙二醛(MDA)含量和抗氧化酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和多酚氧化酶(PPO)]活性的影响,探讨三七细胞膜透性和抗氧化系统酶活性对砷胁迫的响应。结果表明:(1)随着砷质量分数的增加,叶片细胞膜透性和MDA含量逐渐升高;连续2年砷处理为260 mg/kg时,细胞膜透性和MDA含量分别比对照增加160.5%和48.8%;叶片中丙二醛含量与砷质量分数呈极显著正相关关系;(2)第1年砷胁迫,SOD和PPO活性维持稳定,POD活性有所降低;连续2年砷胁迫,SOD活性随着砷质量分数的增加呈现先升高后降低的趋势,且砷处理为260 mg/kg时,酶活性仅为对照的35.7%;POD和PPO活性随砷质量分数的增加而降低;2年砷胁迫CAT活性均随着砷质量分数的增加而增加。总之,在砷胁迫条件下,三七叶片细胞膜受到严重伤害,抗氧化酶系统中POD和PPO活性受到抑制,SOD和CAT是三七叶片中清除过氧化作用的关键酶,连续2年砷胁迫对三七叶片的影响具有一定的累积效应。

Ginsenoside-Rg_6, a Novel Triterpenoid Saponin from the Stem-Leaves of Panax ginseng C. A. Mey.

A novel dammarane-type triterpene oligoglycoside, named ginsenoside-Rg6 3, was isolated from the stem-leaves of Panax ginseng C. A. Mey., together with two known ones, 20(S)-ginsenoside-Rg2 1 and 20(R)-ginsenoside-Rg2 2. On the basis of chemical and physicochemical evidence , the structure of ginsenoside-Rg6 have been elucidated as 6-O-(-L-rhamnosyl-(1?2)-(-D-glucopyranosyl-dammarane-(E)-20(22), 24-diene-3(, 6(, 12(-triol.

Panax notoginseng saponins preconditioning protects rat liver grafts from ischemia/reper- fusion injury via an antiapoptotic pathway

BACKGROUND: Ischemia/reperfusion (I/R) injury is a major cause of primary graft dysfunction and renders an al- lograft more immunogenic in orthotopic liver transplanta- tion (OLT). Panax notoginseng saponins (PNS) has been re- ported to exert protective effects against I/R injury to vari- ous organs. The objective of this study is to investigate whether PNS preconditioning protects rat liver grafts from I/R injury via an antiapoptotic pathway. METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation ( OLT) and were divided into PNS preconditioning group (group P) and normal saline control group (group N) randomly according to whether PNS (50 mg/kg) was injected intra- venously 1 hour before liver grafts harvesting, and sham group (group S). The animals were separately killed 2, 6 and 24 hours after reperfusion. Plasma samples were collect- ed for test of alanine amino-transferase (ALT) and aspartate aminotransferase (AST). Liver tissues were collected to de- tect histological changes, apoptosis and the expression of TNF-α, Bcl-2 and Caspase-3 mRNA. RESULTS: The serum levels of ALT and AST and the apop- tosis index (AI) of liver tissue in group P were lower than in group N significantly 2, 6 and 24 hours after reperfusion. Compared with group N, the expression of TNF-a and Caspase-3 mRNA was reduced significantly in group P 2 and 6 hours after reperfusion and the expression of Bcl-2 mRNA was enhanced significantly in group P 6 and 24 hours after reperfusion. CONCLUSIONS: PNS preconditioning protects liver grafts from I/R injury effectively in rat OLT via an antiapoptotic pathway. The antiapoptotic mechanisms of PNS may in- clude inhibiting the expression of TNF-a and Caspase-3 and enhancing the expression of Bcl-2.

A Novel Hexanordammarane Glycoside from the Roots of Panax notoginseng

Notoginsenoside R10, a novel hexanordammarane glycoside was isolated from the roots of Panax nototginseng, as a minor constituent. Its structure was determined as 6-O-(?-D-glucopyranosyl)-3?,6?,12?-trihydroxy-22,23,24,25,26,27-hexanordammaran-20-one(1), by means of spectroscopic methods.

Saponins from Panax japonicus attenuate age-related neuroinflammation via regulation of the mitogen-activated protein kinase and nuclear factor kappa B signaling pathways

Neuroinflammation is recognized as an important pathogenic factor for aging and related cognitive disorders. Mitogen-activated protein kinase and nuclear factor kappa B signaling pathways may mediate neuroinflammation. Saponins from Panax japonicus are the most abundant and bioactive members in rhizomes of Panax japonicus, and show anti-inflammatory activity. However, it is not known whether saponin from Panax japonicus has an anti-inflammatory effect in the aging brain, and likewise its underlying mechanisms. Sprague-Dawley rats were divided into control groups(3-, 9-, 15-, and 24-month-old groups) and saponins from Panax japonicus-treated groups. Saponins from Panax japonicus-treated groups were orally administrated saponins from Panax japonicus at three doses of 10, 30, and 60 mg/kg once daily for 6 months until the rats were 24 months old. Immunohistochemical staining and western blot assay results demonstrated that many microglia were activated in 24-month-old rats compared with 3-and 9-month-old rats. Expression of interleukin-1β, tumor necrosis factor-α, cyclooxygenase-2, and inducible nitric oxide synthase increased. Each dose of saponins from Panax japonicus visibly suppressed microglial activation in the aging rat brain, and inhibited expression levels of the above factors. Each dose of saponins from Panax japonicus markedly diminished levels of nuclear factor kappa B, IκBα, extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38. These results confirm that saponins from Panax japonicus can mitigate neuroinflammation in the aging rat brain by inhibition of the mitogen-activated protein kinase and nuclear factor kappa B signaling pathways.

Allelochemicals of Panax notoginseng and their effects on various plants and rhizosphere microorganisms

Panax notoginseng(Araliaceae)is an important ginseng herb with various health benefits and a history of cultivation in southwestern China over 400 years.In recent years?.notoginseng has faced serious continuous-cropping obstacles due to its large-scale cultivation.In this study,we aim to explore the allelochemicals of P.notoginseng and their interactions with various plants and rhizosphere microor-ganisms.The chemical constituents of the soil cultivated with 3-year-old P.notoginseng were studied by column chromatography,spectroscopic and GC-MS analyses.We identified 13 volatile components and isolated six triterpenes(1-4,6-7)and one anthraquinone(5).Compounds 1-7 were tested for their effects on seed germination and root elongation in P.notoginseng,corn,wheat,turnip,water spinach and Arabidopsis thaliana.We also examined the effect of compounds 1-7on the growth of ten rhizosphere microorganisms of P.notoginseng.At a concentration of 1.0 ug mL',compounds 3 and 5-7 caused the death of P.notoginseng root cells and compounds 2,6 and 7 induced the death of root cells of A.thaliana.Compounds 1-5and 7 inhibited elongation of A.thaliana root tip cells at a concentration of 10.0μg mL^-1.Moreover,at a concentration of 0.1 mg mL^-1,compounds 3,4,6 and 7 inhibited the growth of probiotics and promoted the growth of pathogens of P.notoginseng.These results suggest that these isolated ursane-type triterpenoid acids and anthraquinone are potential allelochemicals that contribute to continuous-cropping obstacles of P.notoginseng.

Effects of initiating time and dosage of Panax notoginseng on mucosal microvascular injury in experimental colitis

AIM To investigate the effects of Panax notoginseng(PN) on microvascular injury in colitis, its mechanisms, initial administration time and dosage.METHODS Dextran sodium sulfate(DSS)-or iodoacetamide(IA)-induced rat colitis models were used to evaluate and investigate the effects of ethanol extract of PN on microvascular injuries and their related mechanisms. PN administration was initiated at 3 and 7 d after the model was established at doses of 0.5, 1.0 and 2.0g/kg for 7 d. The severity of colitis was evaluated by disease activity index(DAI). The pathological lesions were observed under a microscope. Microvessel density(MVD) was evaluated by immunohistochemistry. Vascular permeability was evaluated using the Evans blue method. The serum concentrations of cytokines, including vascular endothelial growth factor(VEGF)A121, VEGFA165, interleukin(IL)-4, IL-6, IL-10 and tumor necrosis factor(TNF)-α, were detected by enzymelinked immunosorbent assay. Myeloperoxidase(MPO) and superoxide dismutase(SOD) were measured to evaluate the level of oxidative stress. Expression of hypoxia-inducible factor(HIF)-1α protein was detected by western blotting.RESULTS Obvious colonic inflammation and injuries of mucosa and microvessels were observed in DSS-and IA-induced colitis groups. DAI scores, serum concentrations of VEGFA121, VEGFA165, VEGFA165/VEGFA121, IL-6 and TNF-α, and concentrations of MPO and HIF-1α in the colon were significantly higher while serum concentrations of IL-4 and IL-10 and MVD in colon were significantly lower in the colitis model groups than in the normal control group. PN promoted repair of injuries of colonic mucosa and microvessels, attenuated inflammation, and decreased DAI scores in rats with colitis. PN also decreased the serum concentrations of VEGFA121, VEGFA165, VEGFA165/VEGFA121, IL-6 and TNF-α, and concentrations of MPO and HIF-1α in the colon, and increased the serum concentrations of IL-4 and IL-10 as well as the concentration of SOD in the colon. The efficacy of PN was dosage dependent. In addition, DAI scores in the group administered PN on day 3 were significantly lower than in the group administered PN on day 7.CONCLUSION PN repairs vascular injury in experimental colitis via attenuating inflammation and oxidative stress in the colonic mucosa. Efficacy is related to initial administration time and dose.