Botanical,quality control,phytochemistry,pharmacology and toxicity characteristics of Corydalis bungeana Turcz.:a review

Corydalis bungeana Turcz.(CB)is a medicinal herb with significant medicinal value in traditional Chinese medicine.This paper reviews the progress of research on CB’s botany,quality control,phytochemistry,pharmacology and toxicity.The plant’s information was gathered from scientific databases such as PubMed,GeenMedical,Springer Link(https://link.springer.com),Chinese National Knowledge Infrastructure,Pharmacopoeia and Flora.Currently,137 phytochemicals have been identified and isolated from CB,including alkaloids,flavonoids,amino acids,terpenoids,coumarins and organic acids.In addition,many phytochemicals reported various antiinflammatory,antibacterial,antiviral,antitumor,analgesic,hepatoprotective,immunomodulatory,neuromodulatory,and lipid reduction activities.However,the study of its toxicity is still at the preliminary exploration stage and needs further intensive exploration.Herein,we provide an in-depth investigation of the progress of CB to elucidate the underlying mechanisms of activity of CB extracts and its major components,deliver valuable resources and information for further research and rational drug use,and explore the potential research directions and prospects of CB.

Corydalis Rhizoma as a model for herb-derived trace metabolites exploration:A cross-mapping strategy involving multiple doses and samples

Deciphering the metabolites of multiple components in herbal medicine has far-reaching significance for revealing pharmacodynamic ingredients.However,most chemical components of herbal medicine are secondary metabolites with low content whose in vivo metabolites are close to trace amounts,making it difficult to achieve comprehensive detection and identification.In this paper,an efficient strategy was proposed:herb-derived metabolites were predicted according to the structural characteristics and metabolic reactions of chemical constituents in Corydalis Rhizoma and chemical structure screening tables for metabolites were conducted.The fragmentation patterns were summarized from representative standards combining with specific cleavage behaviors to deduce structures of metabolites.Ion abundance plays an important role in compound identification,and high ion abundance can improve identification accuracy.The types of metabolites in different biological samples were very similar,but their ion abundance might be different.Therefore,for trace metabolites in biological samples,we used the following two methods to process:metabolites of high dose herbal extract were analyzed to characterize those of clinical dose herbal extracts in the same biological samples;cross-mapping of different biological samples was applied to identify trace metabolites based on the fact that a metabolite has different ion abundance in different biological samples.Compared with not using this strategy,44 more metabolites of clinical dose herbal extract were detected.This study improved the depth,breadth,and accuracy of current methods for herb-derived metabolites characterization.

Isolation and identification of chemical constituents from the vinegar-prepared Corydalis yanhusuo

Phytochemical investigation of the vinegar-prepared Corydalis yanhusuo led to the isolation of one aristolactam derivative,1,2,8,9-tetramethoxy-5-methyldibenzo[cd,f]indol-4-(5H)-one(1),and seven aporphine alkaloids,including 2,9,10-trimethoxydibenz[de,g]quinolin-7-one(2),1-hydroxy-2,9,10-trimethoxy-7H-dibenzo(de,g)quinoline-7-one(3),oxoglaucine(4),N-methyloxoglaucine trifluoroacetate trifluoroacetate(5),corunine acetate(6),pontevedrine(7),and oxoglaucidaline trifluoroacetate(8).The structures of the isolated compounds were elucidated by extensive spectroscopic data analysis and comparison with the previous reports.Among them,compounds 1 and 2 were obtained as natural products for the first time,and their NMR data were unambiguously assigned.In addition,compound 1 exhibited moderate cytotoxic activity against HepG2 cells with an IC50 value of 16.0±6.4μM.

Corydalis slivensis中的生物碱

作者从保加利亚产的罂粟科植物Corydalis sli-venensis中分离鉴定16种生物碱,其中4种如去氧南天竹啡碱(Dehydronantenine)、紫堇啡碱(Corydine)等由薄层层析检出,2种经光谱鉴定,其余亦均为已知生物碱,如金罂粟碱[(-)-stylopine]、(+)-南天竹啡碱[(+)-nantenine]、普托品(Protopine)和小檗碱等。由光谱法鉴定了(+)-tetrahydrocorysamine(Ⅰ),为无色棱柱形结晶,熔点144℃,(-)-cavidine(Ⅱ)为无色棱柱形结晶,熔点146～147℃二者结构式分别如下:

Study on Quality Standard of Corydalis tomentella Franch

[Objectives] To establish the quality standard of Corydalis tomentella Franch. [Methods] The medicinal materials are identified from the perspectives of morphology,characters,microscopic characteristics and TLC of the plants. The contents of moisture,total ash and water extract are determined. The content of dehydrocavidine is determined by HPLC. The determination is performed on Agilent C18 column with mobile phase consisted of acetonitrile-phosphate buffer solution( containing 20 mmol/L monopotassium phosphate,10 mmol/L diethylamine,0. 1% phosphoric acid)( 28∶ 72,V/V) at the flow rate of 1. 0 m L/min. The detection wavelength is set at 347 nm,and the column temperature is 35℃. The sample size is 10 μL. [Results] The original plants are perennial herbs,and as produced into medicinal materials,they are hairy and shrink into balls with conical principal roots,and obvious cracks on the tubers and roots,curved and fragmented leaves,yellowish white flowers. The quasi-circular pollen grains contain square and columnar crystals,there are substantial nonglandular hairs,and also bordered pits,spiral and reticulate vessels,as well as wood fibers. TLC spots are clear and well-separated. The content of moisture is7. 5%-18. 5%,total ash 20. 5%-26. 2%,and water extract 29. 9%-46. 4%. The linear range of dehydrocavidine is 0. 040 08-2. 404 8 μg( r =0. 999 9); RSD of precision,stability and reproducibility tests is lower than 2. 0%. The recoveries are 95. 6%-102. 5%( RSD =2. 3%,n = 9). [Conclusions] The established standard can be used for quality evaluation of C. tomentella Franch..

Study on Quality Standard of Tibetan Medicine Corydalis dasyptera Maxim.

[Objectives]To establish the quality standard of Tibetan medicine Corydalis dasyptera Maxim.[Methods]According to the research method of drug quality standard in the appendix of 2020 edition of Chinese Pharmacopoeia,8 batches of C.dasyptera Maxim.from different habitats were studied by character identification,microscopic identification and TLC identification.The content of water,total ash,acid-insoluble ash and alcohol-soluble extract was determined,and the content of protopine in medicinal materials was determined by high performance liquid chromatography(HPLC).[Results]The properties and microscopic characteristics of C.dasyptera Maxim.were determined.The TLC characteristic spots of the medicinal materials were clear,the degree of separation was good,and the specificity was strong.Both the test sample and the control sample showed the same yellow-green spots in the corresponding position.It was tentatively determined that the water content of C.dasyptera Maxim.should not exceed 14.0%,the total ash content should not exceed 14.0%,the acid-insoluble ash content should not exceed 3.0%,and the alcohol-soluble extract content should not be less than 18.0%.There was a good linear relationship between the concentration of protopine and the peak area in the range of 16.64-166.40μg·10-3(r=0.9996).The average recovery rate was 98.47%and the RSD was 1.21%(n=6).The content of protopine in 8 batches of C.dasyptera Maxim.was 0.023%-0.093%.[Conclusions]The established quality research method is simple,stable and reliable,and can be used for the quality control of C.dasyptera Maxim.

Antifungal Activity of Corydalis Chloroform Extract against Several Kinds of Pathogenic Fungi of Corn and Soybean

In order to better control common diseases of corn and soybean in Sanjiang plain,the in vitro antifungal activity of corydalis chloroform extract against several common fungal diseases of corn and soybeans was tested using mycelial growth rate method. The test result of mycelial growth rate showed that the inhibition rate of corydalis chloroform extract against Cercosporidium sofinum（ Hara） Liu ＆ Guo and Rhizoctonia solani Kuhn were all above 80%,and EC50 values were52. 98- 58. 98 mg /L. The test result of spore germination showed that the inhibition rates against spore germination of all pathogenic fungi were high,in which the inhibition rates against spore germination of C. sofinum and Bipolaris maydis reached 100%. Biological control test showed that the control effects against other diseases were below 60% except that against R. solani reaching 61. 46%.

An updated classification for the hyper-diverse genus Corydalis(Papaveraceae: Fumarioideae)based on phylogenomic and morphological evidence

The genus Corydalis, with ca. 530 species, has long been considered taxonomically challenging because of its great variability. Previous molecular analyses,based on a few molecular markers and incomplete taxonomic sampling, were clearly inadequate to delimit sections and subgenera. We have performed phylogenetic analyses of Corydalis and related taxa,using 65 shared protein-coding plastid genes from313 accessions(including 280 samples of ca. 226species of Corydalis) and 152 universal low-copy nuclear genes from 296 accessions(including 271samples of Corydalis) covering all 42 previously recognized sections and five independent “series”.Phylogenetic trees were inferred using Bayesian Inference and Maximum Likelihood. Eight selected morphological characters were estimated using ancestral state reconstructions. Results include:(i) of the three subgenera of Corydalis, two are fully supported by both the plastid and nuclear data;the third,subg. Cremnocapnos, is weakly supported by plastid DNA only, whereas in the nuclear data the two included sections form successive outgroups to the rest of the genus;(ii) among all 42 sections and five“series”, 25 sections and one “series” are resolved as monophyletic in both data sets;(iii) the common ancestor of Corydalis is likely to be a perennial plant with a taproot, yellow flowers with a short saccate spur, linear fruits with recurved fruiting pedicels, and seeds with elaiosomes;(iv) we provide a new classification of Corydalis with four subgenera(of which subg. Bipapillatae is here newly described), 39 sections, 16 of which are consistent with the previous classification, 16 sections have been recircumscribed, one section has been reinstated and six new sections are established. Characters associated with lifespan, underground structures, floral spur, fruit and elaiosomes are important for the recognition of subgenera and sections. These new phylogenetic analyses combined with ancestral character reconstructions uncovered previously unrecognized relationships, and greatly improved our understanding of the evolution of the genus.

(+)/(-)-Yanhusuosines A and B, two dimeric benzylisoquinoline-protoberberine alkaloid atropo-enantiomers featuring polycyclic skeletons from Corydalis yanhusuo

(+)/(-)-Yanhusuosines A(1) and B(2), two pairs of trace benzylisoquinoline-protoberberine atropoenantiomeric homodimers featuring an unprecedented 6/7/6/6/6/6 hexacyclic skeleton, were isolated from the tubers of Corydalis yanhusuo. The structures of(+)/(-)-1 and(+)/(-)-2 were elucidated using spectroscopic and quantum-chemical calculation approaches.(+)/(-)-Yanhusuosines A(1) and B(2)represent a new class of alkaloid dimers biogenetically constructed by a molecule of benzylisoquinoline with a unit of protoberberine via an intermolecular [4 + 3] cycloaddition. Their plausible biosynthetic pathways are discussed, and compound 2 exerted moderate inhibitory activity of NO formation in LPS induced RAW264.7 macrophages.

Developing a Novel Single-Marker-Based Method for the Quantitative Evaluation of the Multiple Active Components in Corydalis yanhusuo W. T. Wang

Objective: This study was designed to develop a method for detecting differences in the chemical composition of Corydalis yanhusuo W. T. Wang using high-performance liquid chromatography with a diode array detector technology. Materials and Methods: We established a novel quantitative evaluation method for identifying multiple components in natural extracts using a single-marker method quantitative analysis of multi-components by single marker(QAMS). This method was then validated using eight alkaloid phytochemical markers designed to evaluate C. yanhusuo quality. Results: Our evaluations revealed good linearity(R^(2) ≥ 0.9991) within the range of tested concentrations for all eight alkaloids, with recovery ranging from 95.5% to 101.5%. The evaluations also returned stability results that fell within the acceptable range. Cluster analysis and Heatmap analyses were applied to classify and evaluate alkaloids across 21 different production areas. These results revealed a significant difference in the component profiles between samples from different origins. Conclusions: Thus, these data suggest that in the absence of a material reference, QAMS may help facilitate the stable production of C. yanhusuo. In addition, our data suggest that this method may have value as a promising alternative to common quality evaluations for controlling C. yanhusuo composition.

(+)/(-)-Yanhusamides A-C,three pairs of unprecedented benzylisoquinoline-pyrrole hetero-dimeric alkaloid enantiomers from Corydalis yanhusuo

A chemical investigation on the aqueous extract of Corydalis yanhusuo tubers led to the isolation and structural elucidation of three pairs of trace enantiomeric hetero-dimeric alkaloids,(+)/(-)-yanhusamides A-C(1-3),featuring an unprecedented 3,8-diazatricylco[5.2.2.0^(2,6)]undecane-8,10-diene bridged system.Their structures were exhaustively characterized by X-ray diffraction,comprehensive spectroscopic data analysis,and computational methods.Guided by the hypothetical biosynthetic pathway for 1-3,a gram-scale biomimetic synthesis of(±)-1 was achieved in 3 steps using photoenolization/Diels-Alder(PEDA)[4+2]cycloaddition.Compounds 1-3 exhibited potent inhibition of NO production induced by LPS in RAW264.7 macrophages.The in vivo assay showed that oral administration of 30 mg/kg of(±)-1 attenuated the severity of rat adjuvant-induced arthritis(AIA).Additionally,(±)-1 induced a dose-dependent antinociceptive effect in the acetic acid-induced mice writhing assay.

Alkaloid profiling of the traditional Chinese medicine Rhizoma corydalis using high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry

Since alkaloids are the major active constituents of Rhizoma corydalis(RC),a convenient and accurate analytical method is needed for their identification and characterization.Here we report a method to profile the alkaloids in RC based on liquid chromatography-tandem quadrupole time-of-flight mass spectrometry(LC–Q-TOF-MS/MS).A total of 16 alkaloids belonging to four different classes were identified by comparison with authentic standards.The fragmentation pathway of each class of alkaloid was clarified and their differences were elucidated.Furthermore,based on an analysis of fragmentation pathways and alkaloid profiling,a rapid and accurate method for the identification of unknown alkaloids in RC is proposed.The method could also be useful for the quality control of RC.

Variation in Floral Sex Allocation and Reproductive Success in Sequentially Flowering Inflorescence of Corydalis remota var. lineariloba(Fumariaceae)

In hermaphroditic plants, female reproductive success often varies among different positions within an inflorescence. However, few studies have evaluated the relative importance of underlying causes such as pollen limitation, resource limitation or architectural effect, and few have compared male allocation. During a 2-year investigation, we found that female reproductive success of an acropetally flowering species, Corydalis remota Fisch. ex Maxim. var. lineariloba Maxim. was significantly lower in the upper late developing flowers when compared with the lower early flowers. Supplementation with outcross pollen did not improve female reproductive success of the upper flowers, while removal of the lower developing fruits significantly increased female reproductive success of the upper flowers in both years, evidencing resource limitation of the upper flowers. Female production in upper flowers was greatly improved by simultaneous pollen supplementation of the upper flowers and removal of the lower fruits, suggesting that, when resources are abundant, pollen may limit the female reproductive success of the upper flowers. The less seed mass in the upper flowers didn＇t increase in all treatments due to architecture. In the upper flowers, ovule production was significantly lower and the pollen ： ovule ratio was significantly higher. These results suggest that male-biased sex allocation in the upper flowers may lead to increased male reproductive success, whereas the lower flowers have higher female reproductive success.

Comparative Study for Pharmacological Action of Corydalis Rhizoma before and after Processing

Objective To clarify the rationality and to provide reasonable usage references of processed Corydalis Rhizoma （CR） in clinic by comparing respectively pharmacological actions including analgesic, anti-inflammation, antiplatelet aggregation and spasmolysis of Yuanhu Zhitong Tablet （YZT） and Yanhusuo Decoction （YD） constituted with raw CR （RCR） or processed CR （PCR）. Methods The hot-board, acetic-acid- induced twisting experiments and mouse auricular swelling model by injecting xylene in the abdomen of mouse were adopted to investigate the analgesic and anti-inflammatory effects of the four formulas on Kunming mice; The experiment of contractile activity on isolated rat intestine smooth muscle on Wistar rats was applied to observing the spasmolysis effect of the four formulas; The experiments of platelet aggregation induced by ADP or collagen on Wistar rats by turbidimetry method were used to observe the anti-platelet aggregation effect of the four formulas. Results Compared with RCR in two prescriptions, PCR displayed better analgesic effect （P 〈 0.05, 0.01）, while there was no difference on anti-inflammatory effect; Inhibition of the two prescriptions on smooth muscle spasm was presented well, and the prescription YD with PCR was remarkable （P 〈 0.05, 0.01）; Compared with the blank group, the two prescriptions showed outstanding effects on anti-platelet aggregation （P〈 0.01）, but no difference between RCR and PCR in two prescriptions. Conclusion PCR can improve the analgesic effect of prescription YZT and intensify the spasmolysis effect of prescription YD, which can be explained by the Chinese medicine processing regulation. Therefore, PCR is suggested to be chosen in YZT and YD used to cure pain symptoms.

Effects of Postharvest Processing and Geographical Source on Phytochemical Variation of Corydalis Rhizoma

Objective To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results The results of principal component analysis(PCA) and hierarchical cluster analysis(HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation.Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned.

Metabolomics investigation on antiobesity effects of Corydalis bungeana on high-fat high-sugar diet-induced obese rats

Objective:Corydalis bungeana(CB)is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied as an antiobesity agent.However,the evidence-based pharmacological effects of CB and its specific metabolic alterations in the obese model are not entirely understood.This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats.Methods:A high-fat high-sugar(HFHS)diet was used to induce obese models in rats.CB extract was orally gavaged at 0.18,0.9 and 1.8 g/kg doses for six weeks,and feed intake,body weight,fat pad weight,and blood indexes were measured.Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-fight tandem mass spectrometry(GC-TOF/MS).Results:The results showed that compared with the obese group,the administration of CB extract caused significant decreases in body weight(P<0.05),feed intake,Lee’s index,and perirenal,mesenteric,epididymal fat weight.CB extract also reduced blood triglyceride and total cholesterol levels(P<0.05)of obese rats.Metabolomic findings showed that nine differential metabolites,including pyruvic acid,D-glucuronic acid,malic acid,dimethylglycine,oxoglutaric acid,pantothenic acid,sorbitol acid,fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle,pantothenate and CoA biosynthesis,and glycolysis/gluconeogenesis.Conclusion:This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.

Inhibition of Corydalis decumbens Alkaloids on Hydrogen Peroxide-induced Apoptosis of PC12 Cells through Down-regulating Caspase-3 Expression

Objective To extract alkaloids from Corydalis decumbens(AsCD)by supercritical CO2 fluid extraction(SFE)and to evaluate protective effects of AsCD against hydrogen peroxide(H2O2)-induced apoptosis in rat PC12 cells. Methods AsCD were extracted by SFE and oxidative damage PC12 cells model was induced by H2O2.The survival rate of the cells was determined by MTT assay;Lactate dehydrogenase release was determined by ultraviolet spectrophotometry;Flow cytometry was used to detect apoptosis;Caspase-3 mRNA and protein were determined by real-time PCR and Western blotting assay,respectively.Results AsCD remarkably reduced the cytotoxicity, prevented membrane damage,and inhibited cell apoptosis.AsCD inhibited Caspase-3 mRNA and protein expression induced by H2O2 in PC12 cells.Conclusion AsCD possess protective effects against H2O2-induced apoptosis in PC12 cells,and the mechanism of AsCD responsible to the inhibition of apoptosis is possibly attributed to the down-regulating Caspase-3 expression.AsCD might be useful in the treatment of oxidative stress-related neurodegenerative diseases.

赛北紫堇总生物碱LD_(50)的测定

选取18～22 g KM小鼠90只,给予不同浓度赛北紫堇[Corydalis impatiens(Pall.)Fisch]生物碱灌胃给药,观察30 min内小鼠的存活情况,利用改良寇氏法公式计算LD50。结果表明,赛北紫堇总生物碱的LD50为176 mg/(kg.bw)。为赛北紫堇的开发和利用提供理论支撑。

苦地丁总生物碱对小鼠Ⅱ型胶原关节炎的影响

目的探讨苦地丁对小鼠Ⅱ型胶原关节炎的影响。方法小鼠尾根部皮内注射Ⅱ型胶原乳剂建立小鼠关节炎模型(CIA),苦地丁总生物碱(50,100,200mg·kg-1组灌胃给药,每日1次;模型组和空白组灌胃给予等量蒸馏水;实验结束时考察各组临床积分、发病时间、血清中抗-CⅡ抗体含量、胶原所致耳廓DTH;ELISA法检测PGE2、TNF-α、IL-1的水平。结果苦地丁剂量依赖性延迟CIA小鼠的发病时间,减少关节的肿胀和红斑,缓解CIA小鼠体质量减轻,降低小鼠血清中抗-CⅡ抗体水平,抑制CⅡ诱导的小鼠耳廓迟发型超敏反应;明显抑制巨噬细胞释放肿瘤坏死因子(TNF-α)、白介素1(IL-1),但对前列腺素(PGE2)的产生无明显影响。结论苦地丁对小鼠CIA有明显的抑制作用。