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Objective::To extract crude polysaccharides from Dictamnus dasycarpus (DDP) for separation and purification, and study its anti-psoriasis effect. Method::After interception of DDP with a molecular weight of less than 10 kDa (DDP-UF) using membrane separation technology, four components (DDP-UF-1, DDP-UF-2, DDP-UF-3, DDP-UF-4) were isolated and purified by DEAE-52 cellulose column. Then, the physical and chemical properties and structural characteristics of DDP-UF-1-4 samples were determined by infrared spectroscopy, high performance gel permeability chromatography (HPGPC), high performance liquid chromatography (HPLC) and scanning electron microscope (SEM). Imiquimod cream was selected to induce mouse models of psoriasis, diethylstilbestrol was used to induce vaginal epithelial cell proliferation in female mice, interleulein-17(IL-17) and IL-23 contents of serum in each mouse group were detected by enzyme-linked immunosorbent assay (ELISA), and skin tissues of the mouse back and vaginal epithelial cells had mitotic index changes by hematoxylin-eosin staining (HE). Result::DDP-UF-1-4 all exhibited the characteristic absorption peaks of polysaccharide, and the molecular weights of DDP-UF-1-4 were 10 948, 40 148, 32 222 and 19 943 Da, respectively. The monosaccharide compositions and mole ratios of DDP-UF-1-4 were mannose-glucose-galactose(32.45∶11.35∶8.69), mannose-rhamnose-glucuronic acid-glucose-xylose(25.68∶23.44∶21.62∶18.86∶3.68), mannose-rhamnose-glucuronic acid-galacturonic acid-xylose-galactose(18.68∶4.61∶3.89∶1.65∶5.36∶6.21), glucuronic acid-galacturonic acid-glucose-xylose-galactose(11.63∶15.26∶5.32∶2.08∶3.46), respectively. SEM showed that the morphological structures of DDP-UF-1-4 were flaky or spongy. The drug groups of DDP-UF-1 and DDP-UF-3 improved the skin condition of the psoriasis mice back, inhibited mitosis of female vaginal epithelial cells and significantly reduced the contents of IL-17 and IL-23 in serum (P<0.05, P<0.01). Conclusion::Both DDP-UF-1 and DDP-UF-3 have good anti-psoriasis effects, which may be related to the inhibition of IL-23/IL-17 signaling pathway.
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@#Five compounds were isolated from the petroleum ether extract of Dictamnus dasycarpus using various chromatographic techniques, such as column chromatography over silica gel and Sephadex LH-20. Their structures were elucidated by spectroscopic data(IR, MS, NMR), which were identified as docosanol(1), ethyl 2, 2-dibenzhydryl-3, 3-di-phenylpropionate(2), limonin(3), obacunone(4)and dictamnine(5). Compound 2 was a new compound and compound 1 was isolated from this plant for the first time. PNPG method was used to determine α-glucosidase inhibitory activity of these compounds. The results indicated that compounds 3- 5 possessed stronger inhibitory activities than the positive control of acarbose, which provided a theoretical basis for further development and utilization of Dictamnus dasycarpus.
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OBJECTIVE： To conduct sulfated modification of polysaccharide from Dictamnus dasycarpus （DDP-Ⅲ）， and to compare structure characteristics and anti-psoriasis activity of DDP-Ⅲ before and after sulfated modification. METHODS： DDP-Ⅲ was separated and purified with DEAE-52 anion exchange cellulose column and Sephadex G-100 column. After derived with 1-phenyl-3-methyl-5-pyrazolone， HPLC was used to determine the composition of its monosaccharide. SDDP-Ⅲ was synthesized using esterification reagent （anhydrous pyridine+chlorosulfonic acid） to modify DDP-Ⅲ. The degree of sulfate substitution was determined by barium chloride-gelatin turbidimetric method. The structures were compared by IR， Raman spectrum and SEM before and after modification. The male ICR mice were randomly divided into normal group， model group， positive group （tripterygium glycosides， 20 mg/kg） and DDP-Ⅲ/SDDP-Ⅲ low-dose， medium-dose and high-dose groups （56， 112， 224 mg/kg）. Except that normal group was given vaseline for external use， and other groups were given Imiquimod cream for external use to induce psoriasis model. At the same time， administration groups were given relevant medicine intragastrically 0.4 mL， and both normal group and model group were given constant volume of water intragastrically， once a day， for consecutive 14 d. Two hours after last medication， the serum contents of IL-17 and IL-23 were determined by ELISA. The skin scales near the tail were observed by HE staining， and the number of scales with granular layer was recorded. RESULTS： DDP-Ⅲ was composed of mannose， rhamnose， glucuronic acid， galacturonic acid and glucose. The degree of sulfate substitution was 0.65 for SDDP-Ⅲ. IR and Raman spectrum showed that the characteristic absorption peaks of sulfate radical group appeared near 1 255 cm－1 and 823 cm－1， 1 240 cm－1 and 815 cm－1 for SDDP-Ⅲ， except for same characteristic absorption peak as DDP-Ⅲ. SEM analysis showed that DDP-Ⅲ was flaky， smooth and tightly arranged； SDDP-Ⅲ was massive or granular with porous structure and loose arrangement. Animal experiment showed that compared with normal group， the epidermis of skin lesion was significantly thickened and the granular layer was significantly reduced； serum contents of IL-17 and IL-23 were increased significantly， while the number of scales with granular layer was decreased significantly （P＜0.05 or P＜0.01）. Compared with model group， above symptoms of administration groups were improved to different extent， and serum contents of IL-17 and IL-23 in positive group， DDP-Ⅲ high-dose groups， SDDP-Ⅲ medium-dose and high-dose groups were decreased significantly； the number of scales with granular layer was increased significantly， and above indexes of SDDP-Ⅲ medium-dose and high-dose groups were significantly better than corresponding DDP-Ⅲ group （P＜0.05 or P＜0.01）. CONCLUSIONS： DDP-Ⅲ contains five monosaccharide components such as mannose， etc. Both DDP-Ⅲ and SDDP-Ⅲ possess anti- psoriasis effects， and SDDP-Ⅲ exhibits stronger anti-psoriasis effect than DDP-Ⅲ. Its mechanism may be associated with inhibiting IL-23/IL-17 signaling pathway.
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Objective To obtain the transcriptome dataset of roots of Dictamnus dasycarpus. Methods The root transcriptome dataset of D. dasycarpus was obtained using the high-throughput sequencing platform Illumina HiSeqTM 2000 150PE, followed by systemic bioinformatics analyses. Results A great number of 69 643 286 high quality clean reads were obtained by the transcriptome sequencing analyses. Using Trinity de novo assembling, a total of 49 050 unigenes were finally obtained, with an average length of 841 nt. BLAST analysis indicated that 31 636 (accounting 64.49% of the total unigenes), 22 367 (45.60%), 19 246 (39.23%), 12 595 (25.68%) unigenes were successfully annotated in the NR, Swiss-port, KOG, and KEGG databases, respectively. And GO classification contained the basic three major groups, including biological process, cellular component, and molecular function with 42 subgroups. A total of 132 KEGG standard metabolic pathways were designated, 18 of which were defied as the secondary metabolism. Further analysis revealed that a total of 90 unigenes were involved in the biosynthesis of various alkaloids. Of all unigenes, 1 908 were predicted to have CDS, and 55 families of plant transcription factors were also identified. Using MISA prediction, 4 579 simple sequence repeats (SSRs) were obtained, among which the tri-nucleotide SSRs were abundant with 2 021 (44.1%), whereas the penta-nucleotide SSRs accounted for 3.5%. Conclusion The root transcriptome of D. dasycarpus revealed by the high-throughput sequencing technology will be important for gene functional characterization, secondary metabolism pathway exploration, and regulation mechanism research in this species.
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The root bark of Dictamnus dasycarpus is one of common traditional Chinese medicines (TCMs). Quinoline alkaloids are one of the main active substances in this TCM and possess many biological activities including anti-titumor, anti-inflammation, anti-bacteria, anti-oxidation, and anti-platelet aggregation activities. In this study, eight quinoline alkaloids 1-8 were firstly separated from the root barks of D. dasycarpus. It was difficult to isolate more quinoline alkaloids from the remaining fraction 8 in D. dasycarpus by this conventional chemical separation, so the target analysis method combined LC-MS guided-separation of quinoline alkaloids from fraction 8 was established. MS/MS fragmentation patterns of eight quinoline alkaloids reference standard compounds 1-8 were studied by ultra-performance liquid chromatography-electrospary ionization-mass spectrometry (UPLC-ESI-MS/MS). Based on the feature fragment ion 200, the parent ion scan mode was established for the target analysis of quinoline alkaloids in fraction 8. Finally, 8-methoxyflindersine (9) and N-metilatanina (10) were discovered and isolated quickly from fraction 8 guided by LC-MS, and their structures were identified by NMR and MS. Among them, compound 10 was isolated from the genus Dictamnus for the first time. These results indicated that this method is not only quick and sensitive for analyzing the quinoline alkaloids, but also to effectively guided-separate this kind of alkaloids in the root barks of D. dasycarpus.
Subject(s)
Alkaloids , Chromatography, High Pressure Liquid , Dictamnus , Chemistry , Ions , Phytochemicals , Plant Roots , Chemistry , Quinolines , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
Nineteen compounds, including kihadanin D (1), obacunone (2), kihadanin A (3), kihadanin B (4), kihadanin C (5), limonin (6), evodol (7), fraxinellone (8), furo[2,3-b]quinolin-4-ol (9), preskimmianine (10), ifflaiamine (11), dictamnol (12), naringenin (13), diosmetin (14), wogonin (15), scopoletin (16), cleomiscosin A (17), apocynin (18), and methyl pyroglutamate (19), were isolated from the methanol extract of the root barks of Dictamnus dasycarpus by using various column chromatographies. Their chemical structures were extensively determined on basis of UV, IR, NMR, MS, and CD spectroscopic data analyses. Among them, 1 is a new limonoid, 9 was isolated from plant kingdom for the first time, 11, 13-14 and 17-19 were obtained from the genus Dictamnnus for the first time. Cytotoxicities of compounds 1-18 were tested, and the results indicated that 1 exhibited cytotoxicities against three human cancer cell lines MDA-MB-231, A549 and HT29 with IC₅₈ values of 16.22, 21.72 and 31.06 μmol·L⁻¹, respectively.
Subject(s)
Humans , Cell Line, Tumor , Dictamnus , Molecular Structure , Plant Bark , Plant Extracts , Plant RootsABSTRACT
OBJECTIVE:To study the antioxidant activity of the different polar parts from Dictamnus dasycarpus and its ef-fects on tyrosinase activity. METHODS:Extract was extracted by 95% ethanol from D. dasycarpus,using petroleum ether,chloro-form,ethyl acetate to obtain different polar parts after dissolved in water. 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) method was used to investigate its antioxidant activity [expressed as half inhibitory concentration (IC50)],and tyrosinase method was used to investigate the related activity in different polar parts. RESULTS:The IC50 of petroleum ether,chloroform,ethyl acetate parts for scavenging DPPH free radicals were 0.875,0.824,0.407 mg/mL,respectively. When the mass concentration of each polar part were 25.0,50.0,100,200,300,400,500 μg/mL,the inhibition rate of petroleum ether part to tyrosinase were -3.18%,-4.98%,0.160%,0.044%,2.31%,3.89%,4.29%;that of trichloromethane part were -33.39%,-31.48%,-10.14%,-5.42%,-9.70%,-4.06%,-0.42%;and that of ethyl acetate part were -17.63%,-17.89%,-18.42%,-21.84%,-20.26%,-22.13%,-32.36%. CONCLUSIONS:The capacity in scavenging DPPH free radicals in ethyl acetate part is obvi-ously stronger than the other 2 parts,showing positive correlation with the concentration. Ethyl acetate and chloroform chave an ac-tivation effect on tyrosinase,the activation effect of chloroform part on tyrosinase was negatively correlated with the concentration and petroleum ether part has a two-way regulatory effect on the activity of tyrosinase.
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The present study carried out a phytochemical investigation on the root barks of Dictamnus dasycarpus Turcz, leading to the isolation and characterization of two new aromatic ring butyrolactone derivatives, dasycarpusphenol acid A (1) and dasycarpusphenol acid B (2). Their structures were elucidated by using spectroscopic techniques and HR-FAB-MS. Compounds 1 and 2 exhibited antioxidant activity, with their IC values being 28.95 and 41.76 mg·mL, respectively.
Subject(s)
4-Butyrolactone , Chemistry , Antioxidants , Chemistry , Dictamnus , Chemistry , Molecular Structure , Plant Bark , Chemistry , Plant Extracts , ChemistryABSTRACT
Anticoccidial effects of the root bark of Dictamnus dasycarpus Turcz (Rutaceae) extract (DDE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. Three-day-old chickens (n=30) were assigned to three groups (control, untreated, and DDE 0.1% treated). Chickens were fed a standard diet supplemented with or without DDE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of DDE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The DDE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, DDE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that DDE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of DDE on Eimeria parasites.
Subject(s)
Body Weight , Chickens , Dichlorodiphenyl Dichloroethylene , Dictamnus , Diet , Eimeria , Eimeria tenella , Oocysts , Parasites , RutaceaeABSTRACT
Toxoplasma gondii (T. gondii) causes a life-threatening opportunistic infection. Despite its clinical importance, very few therapeutic drugs against T. gondii are available. Furthermore, these therapeutic regimens are not always suitable for prolonged treatment due to adverse side effects as well as the potential of clinical failure by selecting drug-resistant parasite variants. Dictamnus dasycarpus is known to have many medicinal properties, including anti-inflammatory, anti-fever, and anti-rheumatic activities. In this study, 70% ethanol extract of Dictamnus dasycarpus showed anti-T. gondii effects. Ethanolic extracts of Dictamnus dasycarpus used to treat T. gondii were tested in vitro for their anti-T. gondii activity and cytotoxicity. The selectivity of Dictamnus dasycarpus extract was 7.52, which was higher than that of Sulfadiazine (2.08). We conducted an in vivo animal test to evaluate the anti-T. gondii activity of Dictamnus dasycarpus extract as compared with that of Sulfadiazine. In T. gondii-infected mice, the inhibition rate of Dictamnus dasycarpus extract was high, similar to that of Sulfadiazine. This indicates that Dictamnus dasycarpus extract may be a source of new anti-T. gondii compounds.
Subject(s)
Animals , Mice , Dictamnus , Ethanol , Opportunistic Infections , Parasites , Sulfadiazine , Toxoplasma , ToxoplasmosisABSTRACT
BACKGROUND/AIMS: The aim of this study was to investigate the clinical features of acutely toxic hepatitis associated with ingesting Dictamnus dasycarpus (D. dasycarpus). METHODS: Between January 2004 and July 2009, 28 patients were enrolled in this study. We reviewed the medical records retrospectively. Acutely toxic hepatitis associated with D. dasycarpus was diagnosed by a Roussel Uclaf Causality Assessment Method Values (RUCAM) score of 6 or above. All patients were tested for viral hepatitis A, B, C, cytomegalovirus, and Epstein-Barr virus. Other tests included anti-nuclear antibody, anti-mitochondrial antibody, and anti-smooth muscle antibody. Abdominal pelvic computed tomography was performed. RESULTS: The incidence was female predominant (64% vs. 36%). The mean patient age was 53.0+/-11 years. The symptoms were jaundice (68%), fatigue (57%), nausea (43%), anorexia (43%), and abdominal pain (24%). The mean RUCAM score was 7.0+/-0.8. The biochemical patterns of hepatotoxicity were hepatocellular (n=23, 82%) and mixed types (n=5, 18%). Radiologic findings were as follows: normal findings (29%), lymphadenopathy (50%), edema of the gall bladder wall (46%), periportal edema (43%), splenomegaly (11%), fatty liver (11%), and ascites (7%). The mean hospitalization period was 21.6+/-11.6 days. The mean duration of recovery from hepatitis was 56.6+/-30.4 days, and all patients recovered completely from the toxic hepatitis. One patient who had severe jaundice developed a complication of pure red cell aplasia during the hospitalization period. CONCLUSIONS: The biochemical pattern of liver injury was hepatocellular predominant. Although the initial manifestations and clinical course were variable, all patients completely recovered with supportive care or steroid treatment. Toxic hepatitis was accompanied by pure red cell aplasia in one patient.
Subject(s)
Female , Humans , Abdominal Pain , Anorexia , Ascites , Cytomegalovirus , Dictamnus , Chemical and Drug Induced Liver Injury , Edema , Fatigue , Fatty Liver , Hepatitis , Hepatitis A , Herpesvirus 4, Human , Hospitalization , Incidence , Jaundice , Liver , Lymphatic Diseases , Medical Records , Muscles , Nausea , Red-Cell Aplasia, Pure , Retrospective Studies , Splenomegaly , Urinary BladderABSTRACT
We report a case of fulminant hepatic failure related to ingesting Dictamnus dasycarpus. The patient had taken D. dasycarpus for 8 weeks after boiling down the root of D. dasycarpus in water, to promote health. The main symptoms and signs were general weakness and jaundice. Serology found no evidence of hepatitis A, B, or C infections. Imaging studies, including abdominal ultrasonography and abdominal computed tomography, did not reveal any bile duct structural abnormalities. Based on the RUCAM score (8 points), D. dasycarpus was the probable cause for the drug-induced liver injury. Despite meticulous monitoring and supportive care, the patient died by a progression to fulminant hepatic failure. This case indicates that D. dasycarpus can cause lethal outcomes by fulminant hepatic failure.
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Humans , Bile Ducts , Dictamnus , Chemical and Drug Induced Liver Injury , Hepatitis A , Jaundice , Liver Failure, Acute , WaterABSTRACT
Few cases of hepatitis related to Chinese medicine have been reported in Korea. In such instances, since herbal medicines contain many ingredients, the component that causes the hepatitis to develop is unclear. A 55-year-old man was admitted with sudden-onset jaundice after taking the herbal medicine Dictamnus dasycarpus. The patient had consumed 70 g of alcohol daily and stopped drinking about a month earlier. The physical examination revealed no abnormalities other than scleral icterus. The patient's blood chemistry showed a total bilirubin of 16.0 mg/dL, an ALP of 288 IU/mL, an AST of 1,369 IU/L, and an ALT of 1,660 IU/L. The viral studies were all negative. A liver biopsy showed acute cholestatic hepatitis superimposed on chronic alcoholic fibrosis. This case suggests the development of hepatitis due to an herbal medicine (Dictamnus dasycarpus). Moreover, we recently experienced a case of acute hepatitis that developed after the consumption of Dictamnus dasycarpus.
Subject(s)
Humans , Middle Aged , Alcoholics , Asian People , Bilirubin , Biopsy , Dictamnus , Drinking , Fibrosis , Hepatitis , Herbal Medicine , Jaundice , Korea , Liver , Physical ExaminationABSTRACT
We report four cases of toxic hepatitis that occurred after taking a decoction made by boiling down the root of Dictamnus dasycarpus. The four patients had a median age of 60 years, common symptoms of jaundice and general weakness, and stated that they had not consumed alcohol for at least 5 years. The markers of hepatitis A, B, and C were all negative in the patients. Abdominal ultrasonography revealed common bile ducts with normal diameters. Two of the cases were a mother and daughter, and the other two were sporadic. All of them had consumed a decoction made by boiling down the root of Dictamnus dasycarpus five or six times daily until several days before admission. Dictamnus dasycarpus induced liver injury presenting with a benign course lasting less than 1 month after cessation of the causative agent. We suggest that this natural product can cause acute hepatitis in rural areas.