Research progress on the anti-tumor effect of Codonopsis pilosula

Currently,the mortality rate of malignant tumors ranks second globally,surpassed only by cardiovascular and cerebrovascular diseases.The treatment of malignant tumors poses a formidable challenge to both modern medicine and traditional Chinese medicine(TCM).To date,TCM has developed a substantial foundational theoretical understanding and accumulated significant clinical experience in combating tumors.According to TCM foundational theories,"Qi deficiency"is a critical symptom associated with cancer,and"fortifying the body's vitality while expelling pathogens"is the cornerstone of TCM's approach to tumor treatment and bodily balance.Codonopsis pilosula(CP),a Qi-invigorating herb,is known to invigorat the spleen,benefit the lungs,nourish the blood,and promote bodily fluids.It is often employed as a substitute for ginseng in clinical settings.Prolonged clinical observations have identified key active constituents of CP,such as Codonopsis polysaccharides,isoimperatorin,saponins,lobetyolin,sesquiterpene lactones,and muscone.These ingredients exhibit various therapeutic properties,including anti-tumor,immunomodulatory,anti-infective,antioxidant,and hematopoiesis-enhancing effects.Additionally,when CP is combined with other TCM herbs like Astragalus and Atractylodes macrocephala,it bolsters the body's vital energy and rejuvenates both Qi and blood.CP can be used in combination with chemotherapy agents to mitigate the adverse effects of radiotherapy and chemotherapy.Moreover,CP demonstrates potential in preventing precancerous lesions.This review summarizes recent research findings on the anti-tumor properties of CP,elucidates the anti-tumor effects and molecular mechanisms of its active components,provides a basis for promoting the utilization of CP resources and its active constituents,and offers insights for the research and development of new anti-tumor drugs.

Inspection Methods and Quality Grading Standards for Codonopsis pilosula Seeds

[Objective] This study aimed to investigate the inspection methods and quality grading standards for Codonopsis pilosula seeds. [Method] A total of 27 shares of C. pilosula seeds were collected and surveyed from six counties in Shanxi, Gansu and Shaanxi provinces. The inspection methods of various indicators, such as purity, 1 000-grain weight, germination rate, vitality and moisture content, were investigated in accordance with Rules for Agricultural Seed Testing （GB/T 3543.1-7, 1995）, to develop the appropriate quality inspection method for C. pilosula seeds. Based on the developed inspection method and procedures, the quality of 27 shares of C. pilosula seed samples from different regions was inspected, thus es- tablishing quality grading standards for C. pilosula seeds with K-center clustering method. [Result] C. pilosula seeds are suitable to be cultivated in cool regions. The optimal germination temperature was 25 ~C; the optimal germination bed was gauze bed; the optimal staining conditions were： staining temperature 40 ~, TTC concen- tration 0.5%, staining time 4 h. On the basis of cluster analysis, the quality of C. pilosula seeds could be divided into three grades according to five indicators, includ- ing the purity, 1 000-grain weight, moisture content, vitality, and germination rate. Specifically, vitality and germination rate were important factors of grading standards due to the significant differences, followed by purity, 1 000-grain weight and mois- ture content. [Conclusion] This study provided an important basis for standardized cultivation and industrial development of C. pilosula.

Effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on Sexual Maturity of SD Female Rats

[Objectives] To study the effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on the growth and development of SD female rats. [Methods] The rats were randomly divided into control group,P. ginseng group,C. pilosula group,and P. heterophylla group,intragastrically administered with physiological saline,P. ginseng,C. pilosula,and P. heterophylla for 2 weeks,daily vaginal smear inspection was performed,estradiol( E_2) level before and after intervention was detected,rats were dissected,took out ovary and uterus,and calculated the ovarian index and uterus index. [Results]In 4 groups of rats,there was no significant difference in the time of sexual maturity and ovarian index; compared with the control group,E_2 level of C. pilosula group and P. ginseng group were higher( P < 0. 05);compared with the control group and P. heterophylla group,the ovarian index of P. ginseng group was higher( P < 0. 05); compared with the control group,the body weight of three administration groups increased much more( P < 0. 05). [Conclusions] P. ginseng and C. pilosula may lead to sexual maturity of SD female rats,while P. ginseng,C. pilosula and P. heterophylla can promote increase of body weight of SD female rats.

Protective effect and its mechanism of codonopsis pilosula polysaccharide on memory consolidation disorder induced by cycloheximide in mice

OBJECTIVE To investigate the protective effect of Codonopsis Pilosula Polysaccharide(CPPS)on improving of the memory consolidation disorder induced by Cycloheximide and its possible mechanisms in mice.METHODS The mice was divided into five groups,as normal control group,cycloheximid model group,piracetam positive control group,CPPS 300 mg·kg^(-1) group,and CPPS150 mg·kg^(-1) group.The mice respectively were given saline,piracetam,and CPPS for 15 d.The memory consolidation disorder model in mice was established by ip.Cyclohexylamine,and orally administered CPPS(300 mg·kg^(-1) or 150 mg·kg^(-1))every day.Then experimental groups were subjected Morris Water Maze test.Western blotting analysis were used to analysis the expression of Ca MKⅡ/CREB signaling pathways.RESULTS Morris water maze experiment showed that cyclohexylamine can cause memory consolidation disorder(P<0.01),and giving piracetam and CPPS(300 mg·kg-1)can improve spatial memory impairment in mice(P<0.05,P<0.01).Western blotting experiment results show that compared with normal control group,Ca MKⅡand CREB contents of brain in model group mice had significant decreased(P<0.001);Compared with model group,Ca MKⅡand CREB contents of brain tissue in piracetam and CPPS groups increased significantly(P<0.05,P<0.01,P<0.001).CONCLUSION Cycloheximide can induce the memory consolidation disorder,and its effect in mice related to Ca MK/CREB signaling pathways.CPPS can improved this memory disorder by influence Ca MKⅡ/CREB signaling pathways.

In vitro Propagation of Codonopsis pilosula"Fengdang"

[Objectives]The paper was to establish in vitro propagation of an excellent germplasm of Codonopsis pilosula"Fengdang".[Methods]The effects of different plant growth regulators on callus induction and redifferentiation were tested by using sterile stem explants in vitro that germinated from seeds of superior"Fengdang"plant.[Results]The optimum medium for callus induction was MS+0.5 mg/L NAA+1.0 mg/L 16-BA.The optimal medium for bud differentiation was MS+0.1 mg/L NAA+0.5 mg/L 6-BA,and that for rooting was 1/2 MS+0.2 mg/L NAA+0.2 mg/L IBA.By using the media mentioned above,the rates of callus induction,bud differentiation and rooting reached 91%,100%and 97%,respectively.[Conclusions]The rapid propagation system of regenerated plants in vitro established in this study lays a foundation for the popularization and industrialized seedling of excellent germplasm of"Fengdang".

Effects of Different Seedling Grades on Yield and Commercial Quality of Codonopsis pilosula

[Objectives] This study was conducted to further explore the yield increasing potential of Codonopsis pilosula, and to seek seedlings suitable for efficient production of C. pilosula. [Methods] The effects of C. pilosula seedlings of different sizes on root yield and commercial quality under different cultivation densities were studied. [Results] The fourth-grade seedlings had the least input, the highest yield of 6 948 kg/hm^2, the largest output-to-input ratio at 18.3, the highest root weight proportion of the second-grade product at 58.1%, and the lowest root weight proportion of the first-grade product at 17.6%. Considering the yield, quality and economic benefits comprehensively, the treatment of the fourth-grade seedlings(seedling diameter≤4 mm) is the best choice, and the suitable plant spacing is 8 cm×22 cm. [Conclusions] The study provided a reference basis for the cost saving and efficiency improvement of C. pilosula production.

An inulin-type fructan CP-A from Codonopsis pilosula attenuates experimental colitis in mice by promoting autophagy-mediated inactivation of NLRP3 inflammasome

Inulin-type fructan CP-A,a predominant polysaccharide in Codonopsis pilosula,demonstrates regulatory effects on immune activity and anti-inflammation.The efficacy of CP-A in treating ulcerative colitis(UC)is,however,not well-established.This study employed an in vitro lipopolysaccharide(LPS)-induced colonic epithelial cell model(NCM460)and an in vivo dextran sulfate sodium(DSS)-induced colitis mouse model to explore CP-A’s protective effects against experimental colitis and its underlying mechanisms.We monitored the clinical symptoms in mice using various parameters:body weight,disease activity index(DAI),colon length,spleen weight,and histopathological scores.Additionally,molecular markers were assessed through enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qRT-PCR),immunofluorescence(IF),immunohistochemistry(IHC),and Western blotting assays.Results showed that CP-A significantly reduced reactive oxygen species(ROS),tumor necrosis factor-alpha(TNF-α),and interleukins(IL-6,IL-1β,IL-18)in LPS-induced cells while increasing IL-4 and IL-10 levels and enhancing the expression of Claudin-1,ZO-1,and occludin proteins in NCM460 cells.Correspondingly,in vivo findings revealed that CPA administration markedly improved DAI,reduced colon shortening,and decreased the production of myeloperoxidase(MPO),malondialdehyde(MDA),ROS,IL-1β,IL-18,and NOD-like receptor protein 3(NLRP3)inflammasome-associated genes/proteins in UC mice.CP-A treatment also elevated glutathione(GSH)and superoxide dismutase(SOD)levels,stimulated autophagy(LC3B,P62,Beclin-1,and ATG5),and reinforced Claudin-1 and ZO-1 expression,thereby aiding in intestinal epithelial barrier repair in colitis mice.Notably,the inhibition of autophagy via chloroquine(CQ)diminished CP-A’s protective impact against colitis in vivo.These findings elucidate that CP-A’s therapeutic effect on experimental colitis possibly involves mitigating intestinal inflammation through autophagymediated NLRP3 inflammasome inactivation.Consequently,inulin-type fructan CP-A emerges as a promising drug candidate for UC treatment.

Codonopiloneolignanin A, a polycyclic neolignan with a new carbon skeleton from the roots of Codonopsis pilosula

A neolignan codonopiloneolignanin A（1） with a novel 2,9：20,9：7,70-tricyclo-8,90-neolignane skeleton was isolated from an aqueous extract of the Codonopsis pilosula roots. Its structure including the absolute configuration was elucidated by extensive spectroscopic analysis, including 2D NMR and electronic circular dichroism calculation. The proposed biosynthetic pathway of compound 1 is also discussed.

Validation of Reliable Reference Genes for Accurate Normalization in RT-qPCR Analysis of Codonopsis pilosula

Objective To identify reliable reference genes（RGs） for normalization of real-time PCR（RT-q PCR） data in Codonopsis pilosula. Methods The expression profiles of 10 candidate RGs（GAPDH, ACT, α-TUB, β-TUB, UBQ, CYP, EF-1α, NAC, F-box and PP2A） were examined in C. pilosula during the phenological period. The raw materials examined included roots, stems, leaves, and flower buds at flowering and boll-forming stages, five growth stages of untreated and treated roots with plant growth retardant. The best-suited RGs were accessed using ge Norm, NormF inder, Best Keeper, and Ref Finder algorithms. Results The best-ranked references genes differed across the samples. GAPDH and PP2A were the most suitable for expression analysis in untreated tissues while GAPDH, α-TUB, and PP2A were ranked as the three most stably expressed genes in untreated roots, while NAC and CYP were the most stably expressed genes in stressed（i.e., treated） roots. The expression of UGPase, a key enzyme for CPP biosynthesis, was determined to further validate the selected RGs. Conclusion A total of 10 RGs can be used as reference genes of C. pilosula, however the appropriate one should be used as it may chance.

Targeted isolation and identification of bioactive pyrrolidine alkaloids from Codonopsis pilosula using characteristic fragmentation-assisted mass spectral networking

Codonopsis pilosula(CP),a well-known food medicine homology plant,is commonly used in many countries.In our preliminary study,a series of pyrrolidine alkaloids with high MS responses were detected as characteristic absorbed constituents in rat plasma after oral administration of CP extract.However,their structures were unclear due to the presence of various isomers and the lack of reference standards.In the present study,an MS-guided targeted isolation of pyrrolidine alkaloids of CP extract was performed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC/Q-TOF MS).For data analysis under fast data directed acquisition mode(Fast-DDA),an effective approach named characteristic fragmentation-assisted mass spectral networking was successfully applied to discover new pyrrolidine alkaloids with high MS response in CP extract.As a result,seven new pyrrolizidine alkaloids[codonopyrrolidiums C–I(3–9)],together with two known ones(1 and 2),were isolated and identified by NMR spectral analysis.Among them,codonopyrrolidium B(1),codonopyrrolidium D(4)and codonopyrrolidium E(5)were evaluated for lipid-lowering activity,and they could improve high fructose-induced lipid accumulation in HepG2 cells.In addition,the characteristic MS/MS fragmentation patterns of these pyrrolizidine alkaloids were investigated,and 17 pyrrolidine alkaloids were identified.This approach could accelerate novel natural products discovery and characterize a class of natural products with MS/MS fragmentation patterns from similar chemical scaffolds.The research also provides a chemical basis for revealing in vivo effective substances in CP.

Research on anti-pancreatic cancer mechanism of Codonopsis codonopsis based on network pharmacology

Objective:The mechanism of Dangshen(Codonopsis pilosula)in treating pancreatic cancer(PC)was explored by network pharmacology technology and platform.Methods:The traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)was used to collect the effective compounds and potential targets of C.pilosula,and the genes associated with PC were obtained through the GeneCards database,the interaction genes between the effective compound targets of C.pilosula and PC targets were explored by the Venny method.The following mapping the interaction genes into a protein-protein interaction(PPI)network,and the key targets were screened.Finally,the interactive genes were imported into the DAVID database for gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal enrichment.Results:Twenty-one effective compounds and 98 downstream target genes of C.pilosula were screened through the TCMSP database.A total of 1,278 PC target genes were obtained through the GeneCards database,and the number of overlap genes between C.pilosula targets and PC related genes was 54,of which 10 were key node genes,namely CASP3,TP53,MDM2,AKT1,ESR1,BCL2L1,MCL1,HSP90AA1,CASP9,and CCND.These interactive genes involved a total of 30 typical GO terms and 20 KEGG signals.Conclusion:C.pilosula may play a role in treating PC through multi-component,multi-target,and multi-signal pathways.

Selection and validation of reference genes for quantitative real-time polymerase chain reaction analyses of Serratia ureilytica DW2

Background:Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C.pilosula;nonetheless,until now,no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction(RT–qPCR)data.Methods:To screen stable reference genes of S.ureilytica DW2,the expression of its eight candidate reference genes(16S rRNA,ftsZ,ftsA,mreB,recA,slyD,thiC,and zipA)under different treatment conditions(pH,temperature,culture time,and salt content)was assayed by RT–qPCR.The expression stability of these genes was analyzed using different algorithms(geNorm,NormFinder,and BestKeeper).To verify the reliability of the data,the expression of the glucose dehydrogenase(gdh)gene under different soluble phosphate levels was quantified using the most stably expressed reference gene.Results:The results showed that the zipA and 16S rRNA genes were the most stable reference genes,and the least stable genes were thiC and recA.The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium.Conclusion:Therefore,this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.

Chemical and molecular characterization of Hong Dangshen,a unique medicinal material for diarrhea in Hong Kong

Aim To investigate the plant origin and the identity of the red substance on the surface of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong. Methods The HPLC fingerprints and 5S rRNA gene spacer sequences of Hong Dangshen were obtained and compared with those of genuine species of Radix Codonopsis. The X-ray diffraction spectrum of the red substance was analyzed and compared with that of Halloysitum fingerprints to the Codonopsis species and the highest similarity to Rubrum. Results Hong Dangshen showed very similar HPLC Codonopsis pilosula var. modesta in terms of 5S rRNA gene spacer sequences. The X-ray diffraction spectrum of the red substance was consistent with that of Halloysitum Rubrum. Conclusion The source plant of Hong Dangshen was suggested to be Codonopsis pilosula var. modesta, one of the genuine original plants of Radix Codonopsis （Dangshen） in the China Pharmacopoeia （2005 edition）. The red substance on the surface of Hong Dangshen was indicated to be Halloysitum Rubrum, a traditional medicinal mineral for chronic diarrhea. Our data suggest that Hong Dangshen is derived from the roots of Codonopsis pilosula var. modesta which has been processed with Halloysitum Rubrum, and the name is suggested to be Radix Codonopsis Praeparata Halloysita Rubra.

贵州道真洛党参研究现状与展望

洛党参(Codonopsis pilosula)是川党参在贵州特有的一个生态类型,野生分布于贵州省北部道真自治县高海拔区域(集中分布于贵州大沙河国家级自然保护区内),具体资源量不详,是一种具有潜在药用价值的食补佳品。通过对洛党参药用价值、栽培技术、加工工艺及产业发展等进行综合概述,同时探讨下一步洛党参急需开展研究的方向,为今后研究和开发洛党参提供理论支持。

Investigation on Prescription Screening of a Polyherbal Preparation Based on Splenocyte Proliferation Activity and its Preparation Method

Objective: The present study aims to optimize the extraction conditions of polysaccharides and flavonoids from a polyherbal preparation consisting of three kinds of Chinese medicinal herbs, Codonopsis pilosula, Crataegus pinnatifida, and Lycium barbarum, and evaluation of its immunomodulatory activity in immunosuppressed mice. Materials and Methods: An orthogonal design(L_9 [3]~4) was constructed to achieve the optimal extraction conditions. The immunomodulatory action of the polyherbal preparation was studied at three doses of 10, 20, and 40 mL/kg/day orally by measuring splenocyte proliferation in mice model of cyclophosphamide-induced immunosuppression. Results: The chosen parameters, including the ratio of solvent-to-raw material, duration of extraction, and extraction times, were the fundamental variables that influenced the extract yields. The highest yield of total polysaccharides content was 54.3 mg/mL when the ratio of solvent-to-raw material,duration of extraction, and number of extractions were 12:1, 1.5 h, and 3, respectively. The maximum extraction yield of the flavonoids was 3.5 mg/mL when the ratio of solvent-to-raw material was 12:1, the extraction time was 2 h, and the number of extractions was 3. The prescription screening showed that the impact of the polyherbal preparation on the splenocyte proliferation capacity was more pronounced than its disassembled components. Oral administration of the polyherbal preparation could significantly increase the concanavalin A-stimulated mouse spleen cells proliferation in a dose-dependent manner. Conclusion: These findings suggested that the polyherbal preparation possesses potential for augmenting the immune activity due to the polysaccharide and flavonoid content in these herbal medicines.