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Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.
Subject(s)
China , DNA Barcoding, Taxonomic , DNA, Plant/genetics , Dracaena/genetics , Plants , Resins, Plant , Sequence Analysis, DNAABSTRACT
Ischemia-reperfusion (IR) is one of the significant medical problems in China. Triphenyltetrazolium chloride (TTC) stainingis used to detect the status of the infarct size, and real-time PCR and western blotting are used to detect expressions ofgenes. TUNEL assay has been used to detect apoptosis. Using a tree shrew myocardial IR model, we found that in thereperfusion period, resina draconis (RD) treatment reduced the infarct size by TTC staining, and significantly enhanced thesuperoxide dismutase expression and down-regulated the malondialdehyde concentration in a dose-dependent manner. Inhearts showing IR, Bax was increased and Bcl-2 was reduced, and RD treatment inhibited the IR-induced Bax expressionand up-regulated the IR suppressed level of Bcl-2. TUNEL assay showed that IR induced the apoptosis of myocardial cells,and RD treatment suppressed the IR-induced apoptosis. CHOP and GRP78 were also upregulated in IR hearts, and RDtreatment could significantly attenuate the CHOP and GRP78 levels compared with IR group. We further found that IRdecreased the miR-423-3p expression and upregulated its target gene ERK both in mRNA and protein levels, and RDtreatment upregulated miR-423-3p expression and downregulated ERK expression compared with the IR group. Importantly, miR-423-3p mimics inhibited IR increased ERK, CHOP and GRP78 expressions, and enhanced IR decreased Bcl-2expression, and inhibited the IR-induced apoptosis of myocardial cells. The findings of this study suggest that RD treatmentinhibited the endoplasmic reticulum induced apoptosis of myocardial cells via regulating miR-423-3p/ERK signalingpathway in a tree shrew myocardial IR model.
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Ten fractions(A-J) were prepared by separation of Longxue Tongluo Capsules(LTC) by using silica gel column chromatography and orthogonal experimental design,showing similar chemical profiles with different abundances of peaks.These ten samples were assessed with UHPLC-QE OrbitrapHRMS for 97 common peaks.For the pharmacological activity experiment,three kinds of in vitro cell models including lipopolysaccharide(LPS)-induced BV-2 microglial cells NO release model,oxygen-glucose deprivation/reoxygenation(OGD/R)-treated HUVEC vascular endothelial cells injury model,and OGD/R-treated PC-12 nerve cells injury model were employed to evaluated the bioactivity of each fraction.Based on the contribution of each identified component,grey relation analysis and partial least squares(PLS) analysis were performed to establish component-activity relationship of LTC,identify the potential active components.After that,validation of the potential active components in LTC was carried out by using the same models.The results indicated that 4 phenolic compounds including 7,4'-dihydroxyhomoisoflavanone,loureirin C,4,4'-dihydroxy-2,6-dimethoxydihydrochalcone,and homoisosocotrin-4'-ol,might be the active components for anti-neuroinflammation effect;five phenolic compounds such as 3,5,7,4'-tetrahydroxyhomoisoflavanone,loureirin D,7,4'-dihydroxyhomoisoflavane,and 5,7-dihydroxy-4'-methoxy-8-methyflavane,might have positive effects on the vascular endothelial injury;three phenolic compounds including 5,7,4'-trihydroxyflavanone,7,4'-dihydroxy-5-methoxyhomoisoflavane,and loureirin D,might be the active components in LTC against neuronal injury.
Subject(s)
Humans , Brain Ischemia , Drug Therapy , Capsules , Cell Line , Drugs, Chinese Herbal , Pharmacology , Glucose , Human Umbilical Vein Endothelial Cells , Microglia , OxygenABSTRACT
The research of anti-hepatocellular carcinoma(HCC) drug has attracted more and more attention. Natural products are the important source of active compounds for cancer treatment. A biflavonoid HIS-4 was isolated from Resina draconis in our previous study. MTT assay, hoechst staining, and flow cytometry analysis were used to investigate the effects of HIS-4 on the proliferation and apoptosis of human hepatoma HepG2 and SK-HEP-1 cells. Moreover, the effects of HIS-4 on the migration and invasion ability of HepG2 and SK-HEP-1 cells were evaluated by wound healing assay and Transwell assay. In addition, MTT assay, flow cytometry analyses, Hoechst staining, wound healing assay, Transwell assay, and tube formation assay were used to explore the anti-angiogenic activity of HIS-4 in human umbilical vein endothelial cells(HUVECs). Mechanistically, the HIS-4 regulatory of signal pathways in H9 epG2 and SK-HEP-1 cells were analyzed by Western blot. This results showed that HIS-4 suppressed the proliferation of human hepatoma HepG2 and SK-HEP-1 cells. Moreover HIS-4 induced their apoptosis of HepG2 and SK-HEP-1 cells. HIS-4 inhibited the migration and invasion of HepG2 and SK-HEP-1 cells. Additionally, HIS-4 exhibited angiogenesis effects. Mechanistically, up-regulation of MAPK signaling pathway and down-regulation of mTOR signaling pathway may be responsible for anti-hepatoma activity of HIS-4. Therefore, HIS-4 may be a promising candidate drug for HCC treatment.
Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Biflavonoids , Pharmacology , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Cell Movement , Cell Proliferation , Dracaena , Chemistry , Hep G2 Cells , Liver Neoplasms , Drug Therapy , Pathology , Phytochemicals , PharmacologyABSTRACT
Objective: To rapidly screen the potential analgesic ingredients from Draconis Resina by live cell immobilized chromatography coupled with HRMS. Methods: An HPLC-DAD-ESI-TOF-MS technique was used to rapidly identify the main chemical constituents from Draconis Resina. Based on the bio-specific affinity adsorption of bioactive compounds with receptors or channels on cells, the potential bioactive components in Draconis Resina could be selectively bound to the target cells-mice dorsal root neurons cells, then the chemical constituents with cell target affinity were identified by LC-HRMS. Results: A total of 21 compounds with various structures were tentatively identified and characterized by HPLC-DAD-ESI-TOF-MS, and among them, 10 potentially analgesic active ingredients in Draconis Resina extract combined with dorsal root neurons cells were successfully detected and identified, including two stilbene, two homoisoflavones, one homoisoflavone, two dihydrochalcone, and three flavonoid oligomers. Conclusion: Live cell immobilized chromatography coupled with LC-DAD-HRMS analysis could provide a rapid and efficient tool for finding the potential bioactive components in Draconis Resina for the next pharmacology studies, which provide the reference for exploring of effective materials basis in Chinese medicines.
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Objective To explore the precaution efficacy of Resina Draconis for pressure ulcers in malignant tumor patients with forced position.Methods According to digital table method,168 malignant tumor patients with forced position were randomly divided into observation group and control group.Furthermore,the control group accepted the basic nursing,the observation group accepted the basic nursing and the Resina Draconis packed in bone-like parts.Afterwards,the patients' local skins were observed at intervals,and the incidence rates of pressure sores were compared between the two groups.Results The incidence rate of pressure ulcers of the observation group was 3.57％,which of the control group was 11.90％,the difference between the two groups was statistically significant (x2 =4.085,P =0.043).Conclusion Application of Resina Draconis can prevent pressure sores in malignant tumor patients with forced position.
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The "design space" method was used to optimize the purification process of Resina Draconis phenol extracts by using the concept of "quality derived from design" (QbD). The content and transfer rate of laurin B and 7,4'-dihydroxyflavone and yield of extract were selected as the critical quality attributes (CQA). Plackett-Burman design showed that the critical process parameters (CPP) were concentration of alkali, the amount of alkali and the temperature of alkali dissolution. Then the Box-Behnken design was used to establish the mathematical model between CQA and CPP. The variance analysis results showed that the P values of the five models were less than 0.05 and the mismatch values were all greater than 0.05, indicating that the model could well describe the relationship between CQA and CPP. Finally, the control limits of the above 5 indicators (content and transfer rate of laurine B and 7,4'-dihydroxyflavone, as well as the extract yield) were set, and then the probability-based design space was calculated by Monte Carlo simulation and verified. The results of the design space validation showed that the optimized purification method can ensure the stability of the Resina Draconis phenol extracts refining process, which would help to improve the quality uniformity between batches of phenol extracts and provide data support for production automation control.
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Objective To investigate the therapeutical effect of Resina draconis. on endometriosis rat model. Methods Totally, 60 adult rats were selected and divided into blank control group( n=10) and blood stasis endometriosis model group( n=50 ) . The blood stasis endometriosis model group was randomly divided into 5 groups after modeling: the Resina draconis. high-, medium-, and low-dose group, Guizhi Fuling group and model control group, 10 rats in each group.The Resina draconis. at 0.50, 0.25 and 0.12 g.kg-1 .d-1 was administered to the rats in the Resina draconis. high-, medium-, and low-dose groups, Guizhi Fuling capsule at 0. 25 g . kg-1 . d-1 to those in Guizhi Fuling group, and the same volume of 0. 5% sodium carboxyl methyl cellulose solution to those in model control group and blank control group. All treatments lasted for 3 weeks.At the end of the treatment, the rats were sacrificed;the volume and weight of ectopic tissue were measured. The blood viscosity was detected with blood rheometer.The concentration of estradiol and progesterone was detected using enzyme-linked immune sorbent assay.The matrix metalloprotein-2 expression in ectopic tissues was determined by RT-PCR. Results High, medium and low dose Resina draconis. significantly reduced the volume and weight of the ectopic tissue, and inhibited MMP-2 expression in ectopic tissues.High-, medium-dose Resina draconis. significantly reduced plasma cutting middle and low shear viscosity of the rats.But the Resina draconis. had no obvious effect on serum estradiol and progesterone concentration. Conclusion Resina draconis. reduces blood viscosity and inhibits MMP-2 expression, which can be used for the treatment of blood stasis type of endometriosis.
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Objective: To determine loureirin A and B in different brands of home made resina draconis,providing basis for new quality control method.Methods: A high pressure liquid chromatography(HPLC) method has been developed. UV detector wavelength was set at 270 nm.The mobile phase was acetonitrile acetic acid solution (40∶60).The flow rate was 1.0 ml/min and the column was at room temperature.Results: A good linear correlation was found in the range of 4 to 24 ?g/ml of loureirin A.The regression equation obtained was Y =855.8+803 7.1 X ( r =0.999 9); A good linear correlation was found in the range of 20 to 120 ?g/ml of loureirin B.The regression equation obtained was Y =219.3+808 1.8 X ( r =0.999 9).Conclusion: The quantities of loureirin B in all brands are lower than the limit of quality standard.The quantities of loureirin A are higher than that of loureirin B in the same sample.
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Objective Serum pharmacochemistry was performed to screen the bioactive constituents of Resina Draconis.Methods Based on HPLC fingerprints of Resina Draconis,the migrating constituents absorbed into blood were determined by comparing the HPLC fingerprints of extraction of Resina Draconis(ERB),herb serum sample,and control serum sample,and with the help of LC-MS/MS.Results Six compounds absorbed into blood were detected,five of them were original in form which were 3,4′-dihydroxy-5-methoxystilbene,cochinchinenin B,4′-hydroxy-4,2′-dimethoxy-dihydrochalcone(a new compound),cochinchinenin A,and loureirin B,respectively.The other might be the original constituent or the metabolite.Conclusion The six constituents absorbed into blood are possible bioactive components of Resina Draconis in vivo.Further research will help clarify the bioactive constituents and mechanisms of Resina Draconis.
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Objective:To investigate the effects of Resina Draconis on the levels of plasma glucose,insulin and lipids in normal and diabetic rats. Methods: Hyperglycemia rat models were induced by glucose or adrenaline and rat models with diabetes mellitus were induced by alloxan.The levels of plasma glucose, insulin and lipids (triglyceride, total cholesterol and HDL-C) were determined. Results: Resina Draconis had no obvious effects in decreasing the plasma sugar level in normal fasted rats but could decrease the plasma sugar level and improve the glucose-resistance in glucose-induced or adrenaline-induced hyperglycemic rats and alloxan-induced fasted diabetic rats and increase insulin secretion in both normal and diabetic rats. It could also decrease the plasma lipid level in alloxan-induced diabetic rats. Conclusion: Resina Draconise exerts a good therapeutic effect on hyperglycemia in diabetic rats and its mechanism may be related to the increase of the secretion of insulin.
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To control thor internal quality of Qili Powder, the content of Resina Draconis in this powder was determined with the 2nd derivative peetrograph. This method could eliminate the interference of other components without separation and purification.
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This paper briefly describes China-produced Resina Draconis in respect to the sources of original plant,refining process, chemical compositions,quality analysis and present status of its application in China.