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Objective:To investigate and compare the effects of water extracts of Whitmania pigra Whitman and Hirudinaria ma-nillensis Lesson on the angiogenesis of Tg (kdrl:mCherry) zebrafish. Methods:The zebrafish embryos 6-8 hours after fertilization (6-8hpf) were transferred to the culture medium containing Whitmania pigra Whitman or Hirudinaria manillensis Lesson extract at different concentrations, and the culture medium containing the drugs was replaced every 24 h. And then, at 72 hpf, the larvalmorphology and intersegmental vessels were observed under a microscope. The hatchability of 48-and 72-hpf embryos, and the number of intersegmen-tal vessels and the heart rate of 72-hpf juveniles were measured. Results:Compared with the control group, when the concentration of Whitmania pigra Whitmanis was higher than 30μg· ml-1 , and the concentration of Hirudinaria manillensis Lesson was higher than 20μg· ml-1, the number of intersegmental vessels was significantly reduced (P<0. 01). Compared with the control group, at 48 hpf, when the concentration of the drug groups was higher than 40 μg· ml-1 , the hatchability of the two groups significantly decreased ( P<0. 01);at 72 hpf, the hatchability of Whitmania pigra Whitman decreased significantly at the concentration of 100 μg·ml-1 (P<0.01), while the hatchability of Hirudinaria manillensis Lesson decreased significantly at the concentration of 80 μg· ml-1(P <0. 01). There was no obvious yolk sac edema, pericardial edema and spine curvature in the two groups. The heart rate decreased sig-nificantly (P<0. 01), while was still within the normal range. Conclusion:Both Whitmania pigra Whitman and Hirudinaria manillen-sis Lesson have notable anti-angiogenic activity, and the anti-angiogenesis activity of Hirudinaria manillensis Lesson is stronger. They both have effects on the development of zebrafish embryos, while the toxicity is not obvious.
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Objective:To investigate the enantiomer separation for atropine by capillary electrophoresis .Methods:Capillary elec-trophoresis was used with an elastic quartz capillary column (60 cm ×75 mm, effective length of 40 cm).The concentration of phos-phate buffer was 30 mmol· L-1 .The high and time of injection was 10 cm and 5 s, respectively.The detection wavelength was 225 nm.The best separation conditions were selected including the type and concentration of chiral resolving agent , pH of the buffer solu-tion, operating voltage and organic solvent.Results:The optimum conditions of separation were as follows:the pH of buffer solution was 7.0, the concentration of S-β-CDP was 10 mg· ml-1 , and the operating voltage was 12 kV.Conclusion: The method is simple and fast, which can be used to se parate the optical isomers of atrpo ine.
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Objective To observe the effects of Fuzheng Huayu Capsule on the expressions of the main roles, Rock1 and Rock2, in RhoA/Rock signal transduction pathway in rats with myocardial infarction (MI);To explore the possible mechanism of Fuzheng Huayu Capsule to improve ventricular remodeling in myocardial fibrosis. Methods The MI model was induced by ligation of the left coronary artery. Rats with MI were randomly divided into the model group, Fuzheng Huayu group and Fasudil hydrochloride group. A sham-operation group threading without ligation was setting as a control group, with eight rats in each group. The rats were treated with corresponding medicine for 4 weeks from the second day after modeling. The expression of Rock1, Rock2 and its mRNA were detected by immunohistochemical and real-time PCR method. Results The protein expression of Rock1 and Rock2 in model group were significantly higher than those in the sham-operation group (P<0.05). The protein expression of Rock1 and Rock2 in the Fuzheng Huayu group and Fasudil hydrochloride group were lower than those in the model group. The mRNA expression of Rock2 was significantly higher in the model group than that in the sham-operation group (P<0.05). The mRNA expression of Rock1 in the Fasudil hydrochloride group was lower than that in the model group (P<0.05). The mRNA expression of Rock2 in the Fuzheng Huayu group and Fasudil hydrochloride group was lower than that in the model group (P<0.05). Conclusion Fuzheng Huayu Capsule can decrease the expression of Rock1, Rock2 and Rock2 in the marginal zone of myocardial infarction in rats with MI. The anti ventricular remodeling mechanism of Fuzheng Huayu Capsule maybe related with this.
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Xingnaojing injection has been widely used in the treatment of cerebral vascular diseases, through the following mechanisms: improving the blood-brain barrier (BBB) permeability, anti-oxidant free radical damage, inhibition of excitatory amino acids (EAA) toxicity and calcium overload, inhibition of apoptosis, reducing cerebral edema and inhibition of autophagy. Thus, the paper summarized its progress.
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On the basis of pre-experiment research and the hypothesis of“amputated lumbricus”, this research was aimed to explore mechanism of active components of the amputated lumbricus to promote wound healing. Skin excision was used to establish the mice model. The amputated lumbricus extract was prepared. HE staining and immunohistochemistry techniques were used in the determination of the wound healing rate and changes of VEGF, bFGF, TGF-β1 expression during wound healing period. The results showed that compared with the blank control group, the healing rate of the amputated lumbricus extract group was better. And the HE staining showed better improvement of traumatic tissues. There was no statistic differences on the expression of VEGF and TGF-β1 between the amputated lumbricus extract group and the normal saline group (P> 0.05). The expression of bFGF in amputated lumbricus extract group reached peak earlier than the control group and also lasted a longer time. The amputated lumbricus extract group reached peak on the first day, which had a significant difference (P < 0.05) compared with the control group at the same timepoint. It was concluded that the external application of amputated lumbricus extract had wound healing effect on traumatic skin of mice. Its mechanism may be irrelevant to the expression of VEGF and TGF-β1. However, it may be related to the increasing of bFGF expression in the injured regions during the inflammation stage and proliferation stage.
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Objective To observe the influence of Qizhi Yifei containing serum on the expression of MMP-9 and TIMP-1 mRNA in lung fibroblasts, and explore its mechanism. Methods Trypsin digestion method was used to extract fibroblasts from lung tissue in rats. All fibroblasts were cultured and trained to the fourth generation. Then they were randomly divided into control group, model group and drug serum group. The model group and drug serum group were firstly treated by DMEM with 0.002 5 μg/mL TGF-β1. The control group was treated by DMEM only. The control group and model group were then treated by DMEM with blank drug serum in concentration of 5%, and the drug serum group was treated by DEME with Qiahi Yifei containing serum in same concentration. After 48 and 72 hours, RT-PCR was performed to test the expression of MMP-9 mRNA and TIMP-1 mRNA of each group. Results After 48 hours, MMP-9 and TIMP-1 mRNA were significantly increased in model group and drug serum group compared with control group. There was no difference between model group and drug serum group. After 72 hours, MMP-9 mRNA was up-regulated in model group and was decreased in drug serum group compared with control group. There was no significant difference among the three groups on the expression of TIMP-1 mRNA. Conclusion Qizhi Yifei containing serum can decrease the up-regulated expression of MMP-9 mRNA in lung fibroblasts stimulated by TGF-β1.
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Objective To study the effects of Panax Notoginseng Saponins (PNS) on the damaged expression of ZO-1 of brain microvascular endothelial cells caused by Aβ42.MethodsBrain microvascular endothelial cells were divided into normal control group, model group, PNS low-, medium- and high concentration groups. They were incubated for 24h in 5% CO2 incubator at 37℃ . Then cell vitality was detected by MTT colorimetric method and ZO-1 protein expression was tested by Western blot.Results Stimulation of Aβ42 reduced the activity of microvascular endothelial cells (P<0.01) and suppressed the expression of ZO-1 protein (P<0.01). Compared with the model group, the activity of microvascular endothelial cells of PNS groups, especially the high and medium dose groups (P<0.05), and increased the ZO-1 protein expression. Conclusion PNS can partly recover the barrier function of blood brain barrier through inhibiting the decrease of the activity of microvascular endothelial cells caused by Aβ42.
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This article was aimed to study the effect of pinellia combined with prunella vulgarisin of different ratios on the extraction yield of trigonellin and uracil, in order to investigate the changing disciplines and reasonable com-bination of Shuangxia Soup. A Welch Ultimate AQ-C18 column (4.6 mm×250 mm, 5 μm) was used in the study. The mobile phase, consisting of solvent A (water) and solvent B (acetonitrile) was programmed as a linear gradi-ent. The flow rate was 1.0 mL·min-1. And the detection wavelength was 265 nm. The column temperature was 25℃. HPLC was used in the determination of extraction yield of trigonellin and uracil for the comparison among different compatibilities. The results showed that the linearity was achieved in the range of 0.872-4.36 ng for trigonelline, and 10.8-50.4 ng for uracil. The average recoveries were 96.53%-101.48%, with RSD of 0.18%-4.26%. Com-pared with pinellia, the extraction yield of trigonelline was reduced and the extraction yield of uracil was increased after the compatibility. It was concluded that different compatibilities of Shuangxia Soup had effects on the extrac-tion yield of trigonelline and uracil. The best compatibility ratio still requires in-depth study.
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This article was aimed to study effect of D-ribose on the high-energy phosphate metabolism of skeletal muscle tissues of tired mice. The model was made by burden swimming. And then, the mice were divided into four groups, which were the model group, D-ribose group, caffeine group, and D-ribose with caffeine group). Intragastric administrations of drugs were given to all mice in four groups, three times per day. And all mice continued to swim for three days. The time of swimming was recorded. Gastrocnemius of mice were removed after swimming or 3 days later to measure the concentration of ATP, ADP, AMP and IMP with the HPLC. The results showed that compared with the control group, the time of burden s wimming was significantly prolonged for mice in the D-ribose group and the D-ribose with caffeine group. After three-day recovery, the concentration of ATP, AMP and IMP of gastrocnemius in the D-ribose group and the D-ribose with caffeine group mice was significantly increased. There was no significant difference in the caffeine group mice. It was concluded that D-ribose is involved in the high-energy phosphate metabolism of skeletal muscle tissues of tired mice . D-ribose promotes the recovery of ATP concentration in the gastrocnemius of tired mice, and prolongs the time of burden swimming. Therefore, it has a certain anti-fatigue effect .
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This article was aimed to study the sedative and hypnotic effects of different eluents of Shuangxiatang (SXT). The effects of SXT water decoction, water eluent, 20%, 70% and 95% alcohol eluent on spontaneous ac-tivity and the sleeping induced by subthreshold dose of pentobarbital sodium were measured. The results showed that the SXT decoction, 20% and 95% alcohol eluent can significantly decrease the number of rearing in mice with the percentage of 78.5%, 78.3% and 62.5%, respectively. SXT water eluent and 70% alcohol eluent can significantly decrease the spontaneous activity of mice (P < 0.01), the number of rearing (P < 0.01) and grooming time (P < 0.05). SXT water decoction can significantly shorten sleep latency (P < 0.05), prolong sleep time (P <0.05), and increase rates of sleeping in mice. SXT water eluent can significantly shorten sleep latency in mice (P< 0.05), increase rates of sleeping in mice. SXT water decoction and water eluent have the sedative and hypnotic effects. And the effects are more than alcohol eluents.
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<p><b>OBJECTIVE</b>To study pathological and therapeutical problems concerning myocardial cell mitochondria changes during myocardial cell hypertrophy by culturing rat primary myocardial cells.</p><p><b>METHOD</b>Primary myocardial cells were seperated and cultured together with angiotensin II (Ang II) for 72 or 96 hours. The total protein content with the BCA method and the photography and measurement of cell diameter with inverted microscope reflected myocardial cell proliferation. The mitochondrial membrane potential (Delta Psi m) with fluorescence microscope, the mitochondrial single amine oxidase (MAO) activity with spectrophotometer, the mitochondrial cytochrome oxidase (COX) activity and the injury percentage of mitochondrial outer membrane with microplate reader and the contents of ATP, ADP, AMP with high performance liquid chromatography reflected the injury and energy metabolism of myocardial cell mitochondrial structure and function when being cultured together with Ang II. On that basis, cells were treated with Astragali Radix injection and valsartan for observing pharmacological effects on mitochondrial structure and function in restructured myocardial cells.</p><p><b>RESULT</b>In 72 h and 96 h, compare with the control group, the model group showed significantly increased total protein content and enlarged myocardial cell diameter. During the course of proliferation, the myocardial cell MAO activity and the injury percentage of mitochondrial outer membrane were significantly increased, with significant decrease in mitochondrial COX activity, mitochondrial Delta Psi m and the content of ATP, ADP and rise in the content of AMP. Astragali Radix injection and valsartan reduced myocardial cell total protein content and cell diameter caused by Ang II, decreased myocardial cell MAO activity, significantly increased mitochondrial COX activity and the content of ATP and ADP, and decreased the content of AMP.</p><p><b>CONCLUSION</b>During the process of myocardial hypertrophy, the injury of mitochondrial structure and function and the changes in myocardial cell energy metabolism injury occurred after the injury of mitochondria. Astragali Radix injection and valsartan can reverse myocardial cell mitochondrial structure and function during myocardial cell hypertrophy caused by Ang II. Reversion of myocardial cell hypertrophy and restructuring of myocardial cells helps improve energy metabolism of the myocardial cells.</p>
Subject(s)
Animals , Female , Male , Rats , Astragalus Plant , Chemistry , Cells, Cultured , Drugs, Chinese Herbal , Therapeutic Uses , Hypertrophy , Drug Therapy , Injections , Membrane Potential, Mitochondrial , Mitochondria, Heart , Myocytes, Cardiac , Pathology , Rats, Sprague-DawleyABSTRACT
Objective:Adopting RP-HPLC method,to study and establish the fingerprint of smilacis rhizome.Methods: YMC-Pack ODS-A(5 ?m,4.6 mm?250 mm) chromatographic column,acetonitrile-0.05% phosphoric acid solution gradient elution were applied;with flow rate of 0.8 ml/min and UV detector at 290 nm.Results: Technology investigation indicated that the analytical method this study established has a better reproducibility.The similarity of smilacis rhizoma fingerprints from different sources is better;smilacis rhizome and its three confusions fingerprints are apparently different.Conclusion: HPLC fingerprint analysis can be a method for quality control of medical material of smilacis rhizoma.
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Objective:To establish the quality standard of berberine hydrochloride in Fuyanping capsules by HPLC. Methods:HPLC with Phenomenex C18(250?4.6mm,5?m)column was used.Acetonitrile-0.033mol/l potassium dihydrogen phosphate(30:70)was used as a mobile phase,flow rate was 1.0ml/min and detection wavelength was set at 346nm.Results: Berberine hydrochloride showed a good linear relationship at a range of 0.16-0.83?g,r=0.9999.The average recovery was 99.09%,and RSD was 0.20%.Conclusion:This method is simple,fast and accurate for the quality control of Fuyanping capsules.