ABSTRACT
Objective:To establish an HPLC fingerprint and determine five compounds in Zhengtian pills to provide reference for the effective quality control.Methods:The analysis was carried out on an Agilent XDB-C18analytical column (250 mm × 4.6 mm,5 μm) with gradient elution by acetonitrile (A) -0.19% phosphoric acid solution (B) (0-8 min,10 % A→15% A;10-23 min,15 % A→45 % A;23-50 min,45 % A;50-60 min,45 % A→70 % A;60-70 min,70 % A),the detection wavelength was 310 nm and the flow rate was 1.0 ml/min. The column temperature was 30 ℃. Similarity evaluation was used to evaluate the fingerprints of 12 batches of Zhengtian pills,and five marker components were quantified. Results:There were 33 common peaks in the fingerprints of twelve batches of Zhengtian pills, and five of them (ferulic acid, prim-O-glucosylcimifugin, 5-O-methylvisammioside,imperatorin and isoimperatorin) were identified by comparison with the reference. The similarity of the 12 batches of samples was over 0.99. The linear ranges were 0.050-0.605 μg(r = 0.999 8),0.008-0.100 μg(r = 0.999 2), 0.013-0.150 μg(r = 0.998 6), 0.171- 2.049 μg(r = 0.999 7)and 0.113- 1.352 μg(r = 0.999 6) for ferulic acid, prim-O-glucosylcimifugin,5-O-methylvisammioside, imperatorin and isoimperatorin, respectively. The content of ferulic acid, prim-O-glucosylcimifugin,5-O-methylvisammioside,imperatorin and isoimperatorin was 0.78-0.84 mg/g,0.10-0.13 mg·g-1, 0.18-0.20 mg·g-1,2.44-2.51 mg·g-1and 1.70-1.78 mg·g-1in the 12 batches of samples, respectively. Conclusion:The established method has high sensitivity and specificity,which can be used for the quality control of Zhengtian pills.
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Objective: To investigate the effects of Zhengtian Pills (ZTP) on long term potentiation (LTP) both in Schaffer-CA1 in vitro and perforant path-dentate gyrus (PP-DG) synapses in vivo. Methods: Sprague-Dawley rats were randomly divided into five groups: control, positive control, migraine model, low-, and high-dose ZTP groups. Glyceryl trinitrate (10 mg/kg) was injected subcutaneously to make migraine rat model. Flunarizine (0.9 mg/kg) was set as positive control. Extracellular recording technique in vivo was used to record the effects of ZTP on LTP of PP-DG pathway in anesthetized rats; Using extracellular recording technique in vitro, the effects of ZTP on LTP of Schaffer Collateral-CA1 pathway in rat hippocampal slices were investigated. Results: Compared to the control group, ZTP (1.08 g/kg) significantly enhanced population spike amplitude in PP-DG pathway; Glyceryl trinitrate (10 mg/kg) significantly reduced population spike amplitude in PP-DG pathway; Neither ZTP (0.54 g/kg) nor Flunarizine (0.9 mg/kg) had significant effects on LTP in PP-DG pathway. Compared to the model group, ZTP (1.08 g/kg), ZTP (0.54 g/kg), and flunarizine (0.9 mg/kg) significantly enhanced population spike amplitude in PP-DG pathway. Compared to the control group, ZTP (1.08 g/kg) significantly enhanced field excitatory postsynaptic potential (fEPSP) slope in Schaffer Collateral-CA1 pathway; Glyceryl trinitrate (10 mg/kg) significantly reduced fEPSP slope in Schaffer Collateral-CA1 pathway; Neither ZTP (0.54 g/kg) nor flunarizine (0.9 mg/kg) significant effects on LTP in Schaffer Collateral-CA1 pathway, Compared to the model group, ZTP (1.08 g/kg), ZTP (0.54 g/kg), and Flunarizine (0.9 mg/kg) had significantly enhanced fEPSP slope in Schaffer Collateral-CA1 pathway. Conclusion: Combined with the previous study, the results gave a clue that the effects of ZTP on TRPV1 and hippocampal LTP or their interactions could be the important molecular mechanisms of ZTP acting as migraine and headache medication.
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Objective To observe the action characteristics and comparative advantage of Liujing Toutong Tablets (LTT) about analgesia compared with Zhengtian Pills through different analgesic pharmacodynamic model, experimental migraine model and qi stagnation and blood stasis model. Methods Analgesic effect of LTT on thermal stimulation and chemical stimulation through hot plate test and writhing test caused by acetic acid were observed. Experimental migraine model was induced by nitroglycerin, effect of LTT on the latency and times of scratching head response of migraine model rats were observed, and levels of NO, NOS, and β-EP in serum of migraine model rats were detected. The qi stagnation and blood stasis model was prepared by sc epinephrine hydrochloride combined with ice-water bath, and effect of LTT on cerebral blood velocity and hemorheology of qi stagnation and blood stasis model rats was observed. Results LTT could significantly prolong the latency of hot plate test and decrease writhing times caused by acetic acid in mice. LTT could significantly prolong the latency and decrease the times of scratching head response of migraine model rats, LTT could significantly decrease the level of NO and NOS, increase β-EP obviously in serum of migraine model rats. Also LTT could significantly increase the cerebral blood velocity and decrease blood viscosity as well as plasma viscosity of qi stagnation and blood stasis model rats. Conclusion LTT has significant analgesia effect with characteristics of working early and lasting long on central pain, inflammatory pain and experimental migraine model, also has significant activating blood and dissolving stasis effect on qi stagnation and blood stasis model. The analgesia effect of LTT is equal to Zhengtian Pills, but LTT works early and has multi-target compared with Zhengtian Pills.
ABSTRACT
Objective To investigate the effects and molecular mechanism of Zhengtian Pills (ZTP) on migraine headache. Methods All rats were randomly divided into control, positive control, migraine model, low- and high-dose ZTP groups, and glyceryl trinitrate was injected to induce migraine headache. The time of ears turning red, frequency of scratching head, climbing the cage, and head-twitching were used to evaluate rat behaviors. After 10 d administration of ZTP, the expression levels of transient receptor potential vanilloid 1 (TRPV1) both in cortex and hippocampus were determined by Western blotting. Results After 2 min of glyceryl trinitrate injection, rats showed headache phenomena that parallels the clinical symptoms of migraine, which peaked in 30 min, and lasted for 60 min. Frequency of head-twitching and numbers of scratching head in glyceryl trinitrate (GTN) group were significantly increased. In contrast, after ZTP (1.08 g/kg, ig) treatment, the numbers of scratching head with fore-limb, hind-limb and the frequency of head-twitching were significantly decreased. Flunarizine (FLU) and low-dose ZTP (0.54 g/kg) also showed a trend to decrease the numbers of scratching head and head-twitching frequency, but no significant difference. Besides, ZTP significantly decreased the up-regulated TRPV1 protein expression level both in cortex and hippocampus. Conclusion The present study shows that ZTP could significantly improve the migraine symptoms of headache in rats and TRPV1 might be one of the important molecular mechanisms. This is the first report about the effect of ZTP on TRPV1 protein expression level both in cortex and hippocampus of rats.
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Objective: To investigate the clinical effect of Zhengtian Pills on blood-stasis pattern of migraine with hemodynamics and TCD as markers. Methods: 30 patients with blood-stasis pattern of migraine were given drug Zhengtian Pills for a month. The hemodynamic marker, integrating value of nail fold microcirculation and TCD before and after treatment were compared. Results: There were significant differences in hemodynamic marker, integrating value of nail fold microcirculation and TCD between before and after treatment (P