RESUMO
ObjectiveThe transcriptome characteristics of different tissues of Codonopsis pilosula were analyzed to illustrate the genetic basis of the accumulation of active ingredients in the root of C. pilosula， and to provide theoretical basis for its high-quality production and cultivation. MethodDifferent tissues of C. pilosula at flowering stage were selected as experimental materials， and the contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ were detected by high performance liquid chromatography（HPLC）. RNA-Seq was used to perform transcriptome sequencing of different tissues， and the differentially expressed genes were screened and analyzed by Gene Ontology（GO） and Kyoto Gene and Encyclopedia of Genes and Genomes（KEGG） enrichment analysis， in order to explore the characteristics of active compound distribution and the transcriptional profiles. ResultThe contents of polysaccharides and tangshenoside Ⅰ in the root of C. pilosula were significantly higher than those in other tissues. The transcriptional profiles of the root were significantly different from those of stem， leaf and flower. Cluster analysis， GO and KEGG enrichment analysis of differential gene expression showed that the differential expression genes were mainly enriched in flavonoid and phenylpropanoid biosynthesis， sucrose-starch metabolism， plant hormone signal transduction， plant-pathogen interaction， mitogen-activated protein kinase（MAPK） cascade signal transduction， Adenosine triphosphate（ATP）-binding cassette（ABC） transporter and other pathways. The expression of genes related to biosynthesis of phenylpropanoid compounds were significantly up-regulated in the roots and flowers， and ABC transporter proteins were mostly highly expressed in the flowers. The expression of key enzyme genes for polysaccharide synthesis， such as sucrose：sucrose 1-fructosyltransferase（1-SST） and fructan 1-exohydrolase（1-Feh）， were significantly up-regulated in the roots， and a large number of stress-responsive genes closely related to the accumulation of secondary metabolites were significantly up-regulated in the roots. ConclusionThe active compound content and transcriptional profiles in C. pilosula roots were significantly different from those in stem， leaf， flower and other tissues， showing tissue specificity. Meanwhile， the genes related to stress response and biosynthesis of active compound， such as fructan and phenylpropanoid compounds， were up-regulated in roots of C. pilosula.
RESUMO
OBJECTIVE： To study the tonifying spl een effect of Codonopsis pilosula polysaccharides （CPP） on spleen-deficiency model mice. METHODS ：Totally 60 ICR male mice were randomized into blank group ，model group ，CPP high-dose，medium-dose and low-dose groups [ 1.6，0.8，0.4 g/（kg·d）]，Sijunzi tang （SJZT）group [ 30 g/（kg·d），by crude drug] ， with 10 mice in each group. Except for blank group ，other groups were given Sennae Folium solution intragastrically （0.4 g/d）to establish spleen-deficiency model. After modeling ，administration groups were given relevant drug intragastrically ，and blank group and model group were given 20 mL/kg normal saline intragastrically ，once a day ，for consecutive 6 weeks. After last administration，body weight of mice in each group was measured and their general behavioral characteristics were observed. The red blood cell count in whole blood were recorded ，and the contents of D-xylose，gastrin（GAS），motilin（MTL），somatostatin （SS），vasoactive intestinal peptide （VIP），amylase（AMS），immunoglobulin（IgG and IgM ）and lipopolysaccharide （LPS）in serum were determined ；mRNA and protein expressions of Claudin and Occludin in colon tissues of mice were also detected. RESULTS：Compared with blank group ，body weight of mice in model group was significantly decreased （P＜0.05），and the spleen deficiency symptoms such as sparse back hair and no luster appeared ；the red blood cell count in whole blood ，serum contents of D-xylose，SS，VIP，AMS，IgG and IgM ，mRNA and protein expressions of Claudin and Occludin were significantly decreased（P＜0.05 or P＜0.01）；serum contents of GAS ，MTL and LPS were significantly increased （P＜0.01）. Compared with model group ，the body weight of mice were increased significantly in CPP high-dose group （P＜0.05），and spleen-deficiency symptom was improved significantly. Except for the serum contents of D-xylose and IgM ，the protein expressions of Claudin and Occludin in colon tissue had no statistical significance in CPP low-dose group （P＞0.05），above indexes of other groups w ere improved significantly （P＜0.05 or P＜0.01）. CONCLUSION ：CPP can i mprove spleen-defic iency model mice induced by Sennae Folium，the mechanism of which may be associated with regulating gastrointestinal hormone secretion ， enhancing immunity and protecting intestinal mucosal barrier.