ABSTRACT
Total flavonoids of Dracocephalum moldavica L. (TFDM) is an effective component extracted and isolated from the traditional Uighur medicinal herb Cymbidium fragrans. Cymbidium fragrans has the effects of tonifying the heart and brain, promoting blood circulation and resolving blood stasis, and has been widely used in the treatment of cardiovascular and cerebrovascular diseases for a long time. The purpose of this study was to determine the effect of total flavonoids from Cymbidium fragrans on hypoxia/re-oxygenation (H/R) injury in H9c2 (rat cardiomyocytes) cells and its mechanism. A model (H/R) of hypoxia/re-oxygenation injury in H9c2 cells was established using hypoxia and glucose deprivation for 9 h combined with re-oxygenation and rehydration for 2 h to simulate myocardial ischemia-reperfusion injury. The effects of total flavonoids from Cymbidium fragrans on cell viability, markers of myocardial cell damage, oxidative stress levels, and reactive oxygen radical (ROS) content were investigated, Western blot was used to detect the expression of vascular endothelial growth factor B (VEGF-B) and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathway related proteins. The results showed that the total flavonoids of Cymbidium fragrans significantly increased the viability of myocardial cells after H/R injury, and decreased the content of lactate dehydrogenase (LDH) and creatine kinase isozyme (CK-MB) in the cell supernatant. It significantly reduced malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and decreased intracellular ROS and nitric oxide (NO) content. Western blot analysis showed that the total flavonoids of Cymbidium fragrans decreased Bax levels in H9c2 cells damaged by H/R and increased Bcl-2 expression. Total flavones of Cymbidium fragrans upregulate VEGF-B/AMPK pathway related proteins VEGF-B, vascular endothelial growth factor receptor 1 (VEGFR-1), neuropilin 1 (NRP-1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), phosphorylated adenosine monophosphate activated protein (p-AMPK) and phospho mechanistic target of rapamycin (p-MTOR) levels. The above research results indicate that the total flavonoids of Cymbidium can significantly reduce the H/R injury of myocardial cells, which may be related to the upregulation of VEGF-B/AMPK pathway and inhibition of oxidative stress response.
ABSTRACT
Objective: To investigate the technology for the separation and purification of extract in Dracocephalum moldevica (EDM) by macroporous resin. Methods: Static and dynamic adsorption-desorption were used to select the best one from seven different type macroporous resins; With the content of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid as indexes, the purification technology parameters of EDM were optimized. Results: HPD600 resin showed the best purifying profile, its optimum technology conditions were as follows: The optimum concentration of the sample liquid was 0.08 g/mL equivalent to raw material, the resin column diameter-height ratio was 1:9, the amount of used adsorption was 0.32 g dried medicinal herb/mL resin, sample flow rate was 1.5 BV/h, and adsorption time was 12 h. In the course of elution, the resin column chromatography was eluted with 6 BV of 70% ethanol after removing impurities with 4 BV of water by flow rate of 1.5 BV/h. The contents of total flavonoids, tilianin, luteolin-7-O-β-D-glucuronide, and rosmarinic acid were more than 53%, 5.5%, 4.7%, and 2.5%. Conclusion: Macroporous resin HPD600 is suitable to separate and purify EDM.
ABSTRACT
OBJECTIVE: To establish an HPLC method for simultaneous detemination the contents of rosmarinic acid (phenylpropanoids), luteolin-7-O-β-D-glucuronide and tilianin (flavonoids) in Dracocephalum moldavica L. Extract. METHODS: The samples were separated on Purospher® STARLP RP-18 endcapped column(4.6 mm × 250 mm, 5 μm) by gradient elution with acetonitrile (A)-0.5% formic acid solution(B) (0-28 min, 20% A; 28-55 min, 20%→28% A; 55-80 min, 28%→30% A)as the mobile phase at a flow ratio of 1.0 mL · min-1. The detection wavelength was set at 330 nm and the column temperature was maintained at 35℃. RESULTS: The calibration curves of rosmarinic acid, luteolin-7-O-β-D-glucuronide, tilianin were in good linearity over the ranges of 2.184-21.84 μg · mL-1 (r =0.999 6), 3.14-31.84 μg · mL-1(r =0.999 6), 7.9-39.5 μg · mL-1(r =0.999 5) respectively. The average recoveries were 99.62%, 99.64% and 100.12% with RSD values of 1.27%, 1.05% and 1.09%. CONCLUSION: The method is reliable, simple, and accurate, and can be used for the comprehensive quality control of Dracocephalum moldavica L. extract.
ABSTRACT
Objective: To establish the DNA barcode identification method of Uygur Medicine Dracocephalum mol-davica L. Methods: D. moldavica L. and its ten sibling species of twenty five samples was amplified by ITS2 se-quence, after sequencing and comparing the intraspecific and interspecific variation, we using K2P and NJ meth-ods to analysis the their relationship, then compare the secondary structure. Results: D. Moldavica (KF041160, KF041163, KF041168, KF041169) from Xinjiang without variation in intraspecies, but there are two 2 variation sites in D. moldavica (AY506659) from GenBank. By NJ method, D. moldavica can be distinguished with their sibling species. Also, D. nutans L., D. bipinnatum Rupr. and D. integrifolium Bge. can be distinguished with oth-er sibling species. Conclusion: ITS2 barcode sequence was able to identify D. moldavica and its sibling species, which provides an effective way for the molecular identification of Uygur Medicine D. moldavica.