Mechanism of Hanchuan Zupa Granules in the treatment of Influenza A virus based on network pharmacology and molecular docking / 国际中医中药杂志

Objective:To explore the ingredients, targets, and mechanisms of Hanchuan Zupa Granules in the treatiment of Influenza A virus.Methods:By using Traditional Chinese Medicine Systems Pharmacology Database Analysis Platform (TCMSP), GeneCards, Online Mendelian Inheritance in Man (OMIM), Pharmacogenomics Knowledgebase (PharmGkb), Therapeutic target database (TTD) and DrugBank database to obtain relevant components and targets of Hanchuan Zupa Granules in the treatment of Influenza A virus; R software was used for the obtain of Hanchuan Zupa Granules -Influenza A virus intersection targets; Cytoscape software was applied for the construction of "Hanchuan Zupa Granules-component-target" network; Protein-protein interaction network (PPI) and topological analysis were constructed by STRING database and Cytoscape software. Intersection targets for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were conducted by R software; Auto Dock Tools were used for molecular docking.Results:All together 111 potential active ingredients, with corresponding 131 targetswere identified from Hanchuan Zupa Granules in the treatment of Influenza A virus. Quercetin, apigenin, luteolin, kaempferol, wogonin, etc. are included as core ingredients. STAT3, MAPK1, MAPK3, AKT1, JUN, etc. are included as core targets. Intersection targets were mainly enriched in 178 signal pathways such as IL-17 signal pathway, influenza A signal pathway, TNF signal pathway, etc; Molecular docking showed that core component had a good affinity with the target.Conclusion:Hanchuan Zupa Granules could play the role of anti-Influenza A virus with multi-component-multi-target-multi-pathway,characteristics, and this syudy provide a basis for future experimental research on its mechanism.

Simultaneous Determination of Four Chemical Components in Carthamus tinctorius L. by HPLC / 中国中医药信息杂志

Objective To establish an HPLC method for the simultaneous determination of hydroxysafflor yellow A, rutin, quercetin, and kaempferol in Carthamus tinctorius L..Methods The Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm) was used with diode array detection. The mobile phase was 0.5% phosphoric acid (A) and methanol (B) to separate the four components by gradient elution at 30℃; the flow rate was 0.8 mL/min; the injection Volume was 10 μL; the detection wavelength was at 360 nm.Results The linear ranges of hydroxysafflor yellow A, rutin, quercetin, and kaempferol were 0.0776–0.7760 μg (r=0.9999), 0.0460–0.4600 μg (r=0.9996), 0.0064–0.0640 μg (r=0.9999) and 0.0128–0.1280 μg (r=0.9997), respectively. The average recoveries of four components were 99.68% with RSD=2.29% (n=6), 99.78% with RSD=1.52% (n=6), 102.03% with RSD=1.02% (n=6), 97.03% with RSD=2.05% (n=6), respectively.Conclusion The method is convenient, accurate, sensitive, stable and repeatable, which can be a method for quality control of Carthamus tinctorius L..

Simultaneous determination of gallic acid, methyl gallate and ellagic acid in gall of Quercus infectoria Olivier by HPLC / 国际药学研究杂志

Objective To dertermine the content of gallic acid, methyl gallate and ellagic acid in galls of Quercus infectoria Olivier from different batches by HPLC. Methods The chromatogram colume was Agilent Zorbax SB-C18(4.6 mm × 250 mm, 5μm), gradiently eluting with methanol as mobile phase A and 0.01%phosphoric acid as B at a flow rate of 1 ml/min. The injection Volume was 5 μl. Detection wavelength was at 258 nm. Results Gallic acid was linear in the range of 0.0318-0.1871 g/L with correlation coefficient of 0.9994. The average recovery was 97.52%with RSD 1.41%. Methyl gallate was linear in the range of 0.0769-0.4612 g/L with correlation coefficient of 0.9994. The average recovery was 99.15% with RSD at 1.46%. Ellagic acid was linear in the range of 0.0158-0.0553 g/L with correlation coefficient of 0.9998. The average recovery was 102.75% with RSD at 0.87%. Conclusion The method is convenient, fast, accurate and practicable, and can be used for controling the quality of the galls of Q. infectoria.

Estrogen receptor ERα36 gene silencing impacts the expression of growth-associated protein in PC12 cells / 中华行为医学与脑科学杂志

Objective To investigate the effects of ERα36 (a novel subtype of estrogen receptor alpha) on growth and proliferation in PC12 cells via examining the expression of growth-associated protein in differentiated PC12 cells after ERα36 gene silencing.Methods Transfection of ERα36-shRNA plasmid into PC12 cells was performed to establish the ERoα36 gene silencing cells model (PC12-36L1,PC12-36L2).Immunocytofluorescence was used to examine the expression of ERα36,and Western blot was used to analyze the expression of PCNA,cyclinD1 and MAPK in the PC12 cells.Results ① ERα36 was expressed in both cell types(PC12-36C1,PC12-36L1 and PC12-36L2).Compared with PC12-36C1,PC12-36L2 cells(OD value were respectively 0.95±0.05,0.78±0.10),PC12-36L1 cells significantly decreased expression of ERα36(OD value 0.47±0.12,P＜0.01).② Compared with PC12 and PC12-36C1 cells,PC12-36L1 cells were significantly higher expression of PCNA,CyclinD1 and p-MAPK(P＜0.01)(OD value of PCNA,CyclinD1 and p-MAPK:PC12 cells were respectively 1.00±0.05,1.00± ±0.11,1.00±0.05,PC12-36C1 cells were respectively 1.09±0.15,0.92±0.23,1.12± 0.08,PC12-36L1 cells were respectively 1.74±0.12,2.20±0.25,1.77±0.06).Conclusion ERα36 gene silencing can promote the growth and proliferation in PC12 cells.It suggests that the lower expression of ERα36 may be related to the diseases in nervous system such as brain tumor.

Studies on the Chronic Toxicity of Zukamu Granules in Rats / 医药导报

Objective To study the chronic toxicology of Zukamu granules in rats. Methods A total of 120 healthy SD rats ( male  female = 1 1 ) were randomly divided into the control, low ( 0. 72 g · kg-1 · d-1 ), middle (1.43 g·kg-1·d-1),and high (2.86 g·kg-1·d-1) doses of Zukamu granules. The drug was given orally,once daily for a month,and the controls were given with sodium carboxymethyl cellulose. A 2-week recovery period was allowed after the drug withdrawal. Results Three rats in the high dose group developed diarrhea and loose stools for one day. Compared with the control group,the white blood cells ( WBC) ,red blood cells ( RBC) ,hemoglobin ( HGB) ,hematocrit ( HCT) and chlorine ( Cl-) in the high dose group increased. Prothrombin time ( PT ) , blood urea nitrogen ( BUN ) , alanine aminotransferase ( ALT ) , cholesterol(CHO),and aspartate aminotransferase (AST) decreased markededly in the low,middle and high dose group. No obvious change was found in histopathological examination. Conclusion No obvious toxicity was observed in SD rats treated with Zukamu granules at 1. 4g·kg-1·d-1(equivalently to crude drug of 20. 8 g·kg-1·d-1) given orally for one month.

Determination of Gallic Acid and Kaempferol-3-O-Rutinoside Contents inNymphaea Candida Presl. by HPLC / 中国中医药信息杂志

Objective To establish an HPLC method for the content determination of gallic acid and kaempferol-3-O-rutinoside inNymphaea candida Presl..MethodsThe separation was carried out on a Wondasil C18 column (250 mm×4.6 mm, 5μm). The mobile phase was methanol-0.1% acetic acid solution, with gradient elution;the flow rate was 1.0 mL/min;the detection wavelength was set at 270 nm;the column temperature was 30℃.Results The linear ranges of gallic acid and kaempferol-3-O-rutinoside were 0.065-0.585μg (r=0.999 1), 0.133-1.197μg (r=0.999 5), respectively. The average recoveries of gallic acid and kaempferol-3-O-rutinoside were 102.71%, 102.08%, with RSD of 1.97%, 0.46%, respectively.Conclusion This method was rapid, accurate, and reliable. It can be used for the determination of allic acid and kaempferol-3-O-rutinoside in Nymphaea candidaPresl..

Effects of Alpinia oxyphylla fructus on learning-memory and expression of related signal proteins in hippocampus of brain aging mice / 中华行为医学与脑科学杂志

Objective To investigate the effects and mechanisms of Alpinia oxyphylla fructus (AOF) on learning and memory in D-galactose induced brain aging mice. Methods The brain aging model was induced by s. c D-galactose. Learning-memory ability was tested by passive avoidance test and Morris water maze test, and the expression of synapsin ( Syn), mitogen-activated protein kinase (MAPK) and protein kinase ( PKC ) in hippocampus were examined by Western blot. Results ① Passive avoidance test:the latency in brain aging group( ( 119.80 ±101.80)s) significantly decreased,and the number of errors (4.4 ± 1.3 ) significantly increased compared with the control group( latency: (279.30 ± 31.64) s; number of errors: ( 1. 8 ±0.9), P＜0. 01 ) ). The latency in low dose, middle dose and high dose AOF group( ( 170.25 ± 68.31 ) s, (226.31 ± 73.25 ) s, (263.20 ± 70.55 ) s) significantly increased, and the number of errors in middle dose and high dose AOF group ( ( 2.8 ± 1.2 ), ( 2.3 ±0. 9 ) ) significantly decreased compared with brain aging group (P ＜ 0. 05, P ＜ 0. 0 1 ). ② Morris water maze test:the escape latency in brain aging group was significantly longer, and the time spent in the original quadrant that previously contained the platform was significantly shorter compared with the control group (P＜0. 01 ). The escape latency in 3 AOF groups was significantly shorter (P＜ 0. 05 ), and the time spent in the original quadrant that previously contained the platform in middle and high dose AOF groups was significantly longer compared with brain aging group (P＜0. 05, P＜0. 01 ). ③ Western blot test:the expression of Syn,MAPK and PKC in hippocampus of brain aging group was significantly weakened than that of the control group. In contrast, the expression of Syn,MAPK, PKC were significantly enhanced in all AOF groups. Conclusion AOF could significantly improve the ability of learning and memory in brain aging mice. Its effects might be related to the increase of the expression of Syn, MAPK and PKC in hippocampus.

Effect of water extracts of Alpinia oxyphylla Miq. fruit on memory impairment in cerebral ischemia rats / 中华行为医学与脑科学杂志

Objective This report investigate the intervention effect of the water extract of Alpinia oxyphylla Miq. fruit (AOF)on memory impairment and the mechanism in cerebral ischemia rats. Methods 48 Rats were randomly divided into sham-operated group(n=12), ischemia group(n=12), AOF group Ⅰ( n= 12)and AOF group Ⅱ (n= 12). The model of transient cerebral ischemic/reperfusion was made by bilateral occlusion of common carotid arteries in rats and combination with reducing blood pressure with an abdominal injection of so-dium nitroprusside. Learning-memory ability was observed by step through test. The content of NO and the activi-ties of NOS were measured in hippocampus. Results In step through test,the latency in ischemia group[(143.8±65.2)s]significantly decreased, the number of errors (8.9±4.2 ) significantly increased compared with sham-operated group [latency: (257.2±67.1 ) s; number of errors: (1.7±1.1 ), P<0.01 ]. The latency in AOF group Ⅰ and AOF group Ⅱ[(186.5±46.2) s, (193.4±43.7 ) s ] significantly increased, the number of errors (6.1±2.9,5.2±2.1 ) significantly decreased compared with ischemia group(P<0.05, P<0.01). In hippocampus, the content of NO and the activities of NOS in ischemia group [(56.53±27.42) nmol/mg prot, (17.23±5.64) nmol/mg prot] significantly increased compared with sham-operated group[ (40.02±17.9 ) nmol/mg prot, ( 10.46±6.15)nmol/mg prot], and in AOF group Ⅰ and AOF group Ⅱ [content of NO:group Ⅰ (46.60 ±20. 26)nmol/mg prot,group Ⅱ (42.38±21.23) nmol/mg prot ;activities of NOS:group Ⅰ (13.98±5.13 ) nmol/mg pint,group Ⅱ(13.61±5.27) nmol/mg prot] significantly decreased compared with ischemia group(P<0.05). Conclu-sion AOF could significantly ameliorate the memory impairment in cerebral ischemic rats. Its effects may be in-volved in the decrease of content of NO and activities of NOS in hippocampus.

Progress of Caveolin and Its Role in Brain / 生物化学与生物物理进展

Caveolins are a family of plasmalemmal vesicles caveolae-associated integral membrane proteins and a marker protein of caveolae involved in the formation and localization that associated with vesicular transport, cellular cholesterol homeostasis and signal transduction. Recent years, strong experimental evidences indicated that caveolins play a pivotal role in the brain function such as neural development, synaptic plasticity and neurodegenerative diseases. Recent progress on studies of the structure and functions of caveolins was simply summarized. The regulatory role of caveolins in the brain functions has been reviewed and expected.