Effects and mechanisms of wogonin on airway inflammation in rats with chronic obstructive pulmonary disease / 中国药房

OBJECTIVE To study the effects and potential mechanism of wogonin （Wog） on airway inflammation in rats with chronic obstructive pulmonary disease （COPD）. METHODS Eighty-four rats were randomly divided into control group， model group， Wog low-dose and high-dose groups （intragastric administration of 50， 100 mg/kg）， aminophylline group （positive control， intragastric administration of 2.3 mg/kg）， recombinant rat receptor-interacting protein kinase 1 [rRIPK1， receptor-interacting protein kinase 1 （RIPK1） activator] group （tail vein injection of 8 µg/kg）， and Wog high-dose+rRIPK1 group （intragastric administration of Wog 100 mg/kg+tail vein injection of rRIPK 8 µg/kg）， with 12 rats in each group. Except for control group， COPD model of other groups was induced by smoking combined with tracheal injection of lipopolysaccharide. Twenty-four hours after successful modeling， the rats were administered once a day for 4 weeks. The changes of peak inspiratory flow （PIF）， peak expiratory flow （PEF） and minute ventilation （MV），forced expiratory volume in one second（FEV1）/forced vital capacity（FVC） were measured after the last medication； the serum levels of interleukin 1β（IL-1β）， IL-6 and tumor necrosis factor-α （TNF-α） were measured by ELISA； the pathological changes of lung tissue in rats were observed； the apoptotic rate of pulmonary epithelial cells was detected. mRNA expressions of RIPK1， RIPK3 and mixed lineage kinase domain-like protein （MLKL）， and protein expressions of RIPK1， RIPK3 and p-MLKL were all detected in lung tissue of rats. RESULTS Compared with control group， PIF， PEF， MV and FEV1/FVC of model group were decreased significantly （P＜0.05）， while the levels of IL-1β， IL-6 and TNF- α were increased significantly （P＜0.05）； there was a large number of inflammatory cells infiltration in the lung tissue and bronchialwall thickening in model group； the apoptotic rate of pulmonary epithelial cells，mRNA expressions of RIPK1， RIPK3 and MLKL， protein expressions of RIPK1， RIPK3 and p-MLKL were increased significantly （P＜0.05）. Compared with model group， above indexes of rats were improved significantly in Wog low-dose and high-dose groups （P＜0.05）， and pathological injuries were alleviated significantly. The corresponding indexes of rats were worsened in rRIPK1 group （P＜0.05）， and pathological damage had further worsened. rRIPK1 significantly attenuated the inhibitory effect of high-dose Wog on airway inflammation and RIPK1/RIPK3/ MLKL pathway in COPD rats （P＜0.05）. CONCLUSIONS Wog may improve airway inflammation in COPD rats by inhibiting RIPK1/RIPK3/MLKL signal pathway.

Study on mechanism difference of baicalein and wogonin inhibiting energy metabolism of hepatoma cells / 中国药房

OBJECTIVE To explore the difference in th e mechanis m of baicalein and wogonin inhibiting the energy metabolism of hepatoma cells. METHODS Human hepatoma HepG 2 cells were divided into blank control group （without medicine），different dose groups of baicalein and wogonin （1.25，2.5，5，10 and 20 μmol/L）. The effects of baicalein and wogonin on the viability of HepG 2 cells were detected by MTT assay. HepG 2 cells were divided into blank control group （without medicine），baicalein group and wogonin group. After administration ，the concentration of ATP in cell was detected by enhanced ATP kit. The levels of cell glycolysis and mitochondrial energy metabolism were evaluated by glycolysis and mitochondrial pressure test kit ；the affinity of baicalein and wogonin with key enzymes of energy metabolism was predicted by molecular docking ，and the key enzymes of energy metabolism with high affinity were screened ；the expression of key enzymes of energy metabolism was detected by Western blot. RESULTS Within the dose range of 2.5-20 μmol/L，the half inhibitory concentrations of baicalein and wogonin were 12.84 and 24.09 μmol/L；baicalein 1.25 μmol/L and wogonin 2.5 μmol/L had no effect on cell viability ，so it was selected as the dosage for subsequent experiments. Compared with blank control group ，the concentration of ATP in HepG 2 cells decreased significantly in baicalein group and wogonin group （P＜0.05）；the inhibitory effects on basic acidification rate of HepG 2 cells in wogonin group were significantly stronger than those of baicalein group （P＜0.05），but there was no significant difference between them on the basic oxygen consumption rate （P＞0.05）；baicalein had strong binding to pyruvate kinase M 2 and mitochondrial enzyme complexes Ⅰ（CⅠ），C Ⅱ and C Ⅳ，while wogonin only had strong binding to pyruvate kinase M 2； wogonin could significantly down-regulate the protein expressions of hexokinase ，phosphofructokinase，pyruvate kinase M 2，CⅠ， C Ⅱ and C Ⅳ（P＜0.05），but there was no statistical significance in the effect of baicalein on the regulation of these enzymes （P＞ 0.05）. CONCLUSIONS Both baicalein and wogonin can inhibit the energy metabolism of hepatoma HepG 2 cells，but the mechanism is different ：the effect of baicalein is related to the activity of key enzymes ，while the effect of wogonin is related to the inhibition of the expression of key enzymes of energy metabolism.

Prediction and analysis of quality markers between crude and wine-processed Scutellaria baicalensis based on "components-targets-metabolism" network analysis / 中草药

Objective: To explore the potential Q-markers between crude Scutellaria baicalensis (CSR) and wine-processed S. baicalensis (WSR) based on "components-targets-metabolism" network analysis. Methods: According to the differential components between CSR and WSR, the network relationship of "components-target-metabolomics" was constructed combining network pharmacology and metabolomics. The correlation analysis was then conducted between flavonoids glycosides, and aglycones in S. baicalensis, between differential components and endogenous metabolites to predict the potential quality markers. Results: In this study, combining the results of network pharmacology and metabolomics, baicalin and oroxylin A-7-O-glucuronide were regarded as the quality markers of CSR; Baicalein and wogonin were considered as the quality markers of WSR. Conclusion: It is crucial wine-processed mechanism of S. baicalensis that glutinous rice wine can promote the dissolution and absorption of aglycones. Overall, identification of the differences between Chinese herbal decoction pieces and it processed product, combining analysis of network pharmacology and metabolomics, which provides a demonstration for the investigation of quality markers of Chinese herbal pieces.

Simultaneous determination of baicalein and wogonin in Baipuhuang tablets by HPLC / 国际药学研究杂志

Objective: To establish an HPLC method for simultaneous determination of baicalein and wogonin in Baipuhuang tablets. Methods: HPLC was performed on an Agilent C 18 column(4.6 mm×250 mm, 5 μm)using the gradient acetoni trile(A)-water(B)at the 1.0 ml/min flow rate as mobile phase. The columm temperature was 30℃ and the detection wavelength was 280 nm. Results: The linear regression equation of baicalein was Y=24117X-223.95(r=0.9990);the linear range was 0.02~0.2 μg with the average recovery of 98.95% and the RSD of 0.64%(n=9). The linear regression equation of wogonin was Y=16237X-26.698(r=0.9996);the linear range was 0.02~0.2 μg with the average recovery of 99.28% and the RSD of 1.03%(n=9). The RSD values for the precision, stability and repeatability were all less than 2%. Conclusion: The established HPLC method appears to be highly sen sitive, accurate and reproducible, which could be used for the quality control of Baipuhuang tablets.

Inhibitory effect of flavonoids from Scutellariae Radix on human cytochrome P450 1A / 中国中药杂志

This study investigated the inhibitory effect of eight natural flavonoids in Chinese herb Scutellariae Radix on huamn cytochrome P450 1 A(CYP1 A), a key cancer chemo-preventive target. In this study, phenacetin was used as a probe substrate for CYP1 A, while human liver microsomes and recombinant human CYP1 A enzymes were used as enzyme sources. Liquid chromatography-tandem mass spectrometry was used to monitor the formation rates of acetaminophen, the O-deethylated metabolite of phenacetin. The dose-dependent inhibition curves were depicted based on the changes of the formation rates of acetaminophen, while the IC_(50) were determined. Inhibition kinetic analyses and docking simulations were used to investigate the inhibition modes and mechanism of wogonin(the most potent CYP1 A inhibitor in this herb), while the inhibition constants(K_i) of wogonin against both CYP1 A1 and CYP1 A2 were determined. Among all tested flavonoids, wogonin, 7-methoxyflavanone and oroxylin A displayed a strong inhibitory effect on CYP1 A(IC_(50)100 μmol·L~(-1)). Further investigations demonstrated that wogonin had a weak inhibitory effect on other human CYP enzymes, suggesting that it could be used as a lead compound for the development of specific inhibitors of CYP1 A. Furthermore, the inhibition kinetic analyses clearly demonstrated that wogonin could strongly inhibit phenacetin O-deethylation in both CYP1 A1 and CYP1 A2 in a competitive manner, with K_i values at 0.118 and 0.262 μmol·L~(-1), respectively. Molecular docking demonstrated that wogonin could strongly interact with CYP1 A1 and CYP1 A2 via hydrophobic and π-π interactions, as well as Ser120 and Ser116 in CYP1 A1 via hydrogen-bonding. In conclusion, this study found that some flavonoids in Scutellariae Radix displayed a strong inhibitory effect on CYP1 A, while wogonin is the most potent CYP1 A inhibitor with a relatively high selectivity towards CYP1 A over other human CYPs.

Fingerprinting study on Niuhuang Shangqing Pills based on comprehensive two-dimensional liquid chromatography / 中草药

Objective: To develop a new three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using Niuhuang Shangqing Pills (NSP) as an example. The developed method could offer new method for the quality control of NSP. Methods: In first dimension, the separation was achieved with an Acquity UPLC HSS CYANO column (100 mm × 2.1 mm, 1.8 μm), methanol-0.1% formic acid in water were used as mobile phases, flow rates were 0.1 mL/min. In second dimension, the separation was achieved with a Kinetex Phenyl-Hexyl column (50 mm × 3 mm, 2.6 μm), acetonitrile-0.1% formic acid in water were used as mobile phases, flow rates were 1.5 mL/min, detection wavelength was set at 254 nm, and acquiring frequency was at 12.5 Hz. Column temperature for each dimension was 40 ℃ and volume of loop linking the two dimensions was 100 μL. Three similarity-calculating Methods:, Euclidean Distance, Cosine, and Correlation Coefficient, were employed to assess the similarities among the 21 samples on the market using medians with arithmetic means of peak volumes of the common peaks as control fingerprints. Results: The three-dimensional fingerprints of 21 batches of NSP samples on the market were developed; Eighteen common peaks were assigned and five of them were identified, which were geniposide (1), pulegone (8), baicalin (9), imperatorin (15), and wogonin (16). Conclusion: A three-dimensional fingerprinting method and its assessing Methods: based on comprehensive two-dimensional liquid chromatography using NSP as an example were successfully developed for the first time, suggesting that it is a feasible method for developing fingerprints for Chinese materia medica. This work improves and supplements the traditional liquid-chromatography fingerprints.

Effects of ecological factors and gene expression of key enzymes on synthesis of major medicinal ingredients of Scutellaria baicalensis in autumn / 中草药

Objective:To study the dynamic changes of the pharmacodynamic components of Scutellaria baicalensis in the harvest period and the effects of ecological factors and key enzyme expression on it. Methods :The artificial cultivated annual S. baicalensis was studied and the expression of nine key enzyme genes (PAL, C4H, 4CL, CHS, CHI, FNS, F6H, UBGAT, and GUS) in the roots of S. baicalensis were determined by real-time quantitative PCR. The content of four main flavonoids (baicalin, wogonoside, baicalein, and wogonin) in the roots was determined by HPLC. The meteorological data of S. baicalensis were collected by the ecological meteorological station. SPSS statistical software and DPS statistical software were used for data analysis. Results: The content of flavonoids of four monomers of annual S. baicalensis decreased slowly in autumn, so the best harvest time of S. baicalensis was in early September. The results of grey correlation analysis showed that the ecological factors affecting the four flavonoids were SWC, Max Ta, RH, and PAR. The expression of C4H and UBGAT genes had an important effect on the accumulation of flavonoids in the roots of S. baicalensis in autumn. Maximum rainfall intensity may indirectly affect the accumulation of the pharmacodynamic components of S. baicalensis by affecting the gene expression of key enzymes. Conclusion: The dynamic changes of four main flavonoids of annual S. baicalensis in autumn and the expression of key enzyme genes of S. baicalensis annual are clarified, providing the theoretical basis for the clarification of the physiological and ecological mechanism of the biosynthesis of S. baicalensis and the improvement of the quality of S. baicalensis.

Research progress on pharmacological effects and mechanisms of flavonoids from Scutellaria baicalensis based on aging hypotheses / 中草药

Baicalin, baicalein, wogonoside, and wogonin are major flavonoids of Scutellaria baicalensis, which have been reported to possess various pharmacological effects such as anti-oxidation, immunomodulation, mitochondrial protection, telomerase inhibition, and anti-inflammatory activities. Recently, researches on their anti-aging activity have also gradually increased. Therefore, combining with kinds of aging hypotheses, e.g. the free radical aging hypothesis, immunosenescence aging hypothesis, spleen-kidney aging hypothesis of Chinese medicine, the mitochondrial aging hypothesis, the telomere hypothesis of cellular aging, inflamm-aging, we focus on the related pharmacological properties and mechanisms of these four flavonoids and make a review, aiming to provide a theoretical basis for the anti-aging research of flavonoids in S. baicalensis.

Network pharmacological analysis of Chaihu Guizhi Decoction for treating different diseases with same method on gastric ulcer and epilepsy / 中草药

Objective: Under the guidance of traditional Chinese medicine theory, this paper used network pharmacology model to explore the effective components and mechanism of “treating different diseases with same method” of Chaihu Guizhi Decoction (CHGZD) in treating gastric ulcer and epilepsy, which provided modern medical evidence for the theory of “treating different diseases with same method” of traditional Chinese medicine. Methods: The chemical constituents and potential targets of CHGZD were collected by TCMSP and TCMID databases. Disease targets of gastric ulcer and epilepsy were obtained in PubMed, CTD, and OMIM databases. Cytoscape 3.6.0 software was used to map CHGZD for gastric ulcer and epilepsy. The pharmacodynamic basis and molecular mechanism of the “treating different diseases with same method” network, and the functional and pathway enrichment analysis of gene targets was analyzed by DAVID 6.8 and KOBAS 3.0. Results: A total of 198 kinds of chemical constituents, 417 target targets, 114 gastric ulcer disease targets, and 461 epileptic disease targets were collected from CHGZD. Network analysis showed that 152 active components such as quercetin, beta-sitosterol, wogonin, kaempferol, and saikosaponin a in CHGZD played a role in the treatment of gastric ulcers and epilepsy through 17 common targets including PTGS2, VEGFA, TP53, IL6, and TNF, and 62 pathways such as pathways in cancer, and proteoglycans in cancer. Conclusion: The similar efficacy network composed of 17 common targets in gastric ulcer and epilepsy was the basis for CHGZD to play the role of “different disease and common treatment”. A total of 152 components work together through 62 pathways and 17 pathways to achieve the effect of “treating different diseases with same method”, which provides modern medical evidence for the traditional Chinese medicine to treat gastric ulcer and epilepsy from the liver and spleen.

Study on efficacy network of Chaihu Guizhi Ganjiang Decoction in treating insomnia / 中草药

Objective: To investigate the mechanisms of effects of Chaihu Guizhi Ganjiang Decoction in the treatment of insomnia by using network pharmacology methods. Methods: TCMSP and TCMID were used to lock the targets of seven herbs in Chaihu Guizhi Ganjiang Decoction. TTD, DrugBank, and PubMed were used to search targets of insomnia and construct a “disease-prescription-target” network. STRING and Cytoscape were used to perform enrichment analysis and clarify the mechanism of core targets in the network. Results: The PPI network of Chaihu Guizhi Ganjiang Decoction contained 640 targets and the PPI network of insomnia included 175 targets. A total of 29 core targets and 80 interactions were found after enrichment analysis between two PPI networks. After GO enrichment analysis and KEGG pathway analysis of 29 key targets, we found that 171 active ingredients in Chaihu Guizhi Ganjiang Decoction such as saikosaponin a, saikosaponin d, quercetin, calcium carbonate, 6-gingerol, kaempferol, and wogonin, which played a role in the treatment of insomnia mainly through 29 core targets such as CACNA1C, GABRA1, GABRA2, GABRB3, GABRA3, with biological processes such as target and synaptic signaling, regulation of membrane potential, G-protein coupled receptor signaling pathway, and molecular functions such as neurotransmitter receptor activity, ion-gated channel activity, GABA-A receptor activity, and functional pathways composed by plasmalemma, synapse, and other cells such as neural active ligand-receptor interaction, retrograde endogenous cannabinoid signal transduction, and serotoninergic synapses. Conclusion: The pharmacological substance basis for the treatment of insomnia was composed of 171 active ingredients such as saikosaponin a and saikosaponin d. The efficacy network of “soothing liver and invigorating spleen, regulating yin and yang” was constituted by several pathways like the neural active ligand-receptor interaction and 29 targets such as CACNA1C. Our results provide network pharmacological evidence for clinical rational use of Chaihu Guizhi Ganjiang Decoction for insomnia.

Simultaneous Determination of 6 Components in Chaihuang Tablets by HPLC-DAD / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of 6 components in Chaihuang tablets， such as baicalin， wogonoside， baicalein， wogonin， saikosaponin a and saikosaponin d in Chaihuang tablets. METHODS： HPLC-DAD method was used to detect 3 batches of Chaihuang tablets from same manufacturers. The determination was performed on Agilent Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-triethylamine phosphate aqueous solution （pH adjusted to 7.0， gradient elution） at flow rate of 1.0 mL/min. The detection wavelengths were set at 210 nm （saikosaponin a， saikosaponin d） and 277 nm （baicalin， wogonoside， baicalein， wogonin）. The column temperature was 30 ℃， and sample size was 5 μL. RESULTS： The linear ranges of baicalin， wogonoside， baicalein， wogonin， saikosaponin a and saikosaponin d were 0.379 5-7.590 4 μg，   0.082 96-1.659 2 μg， 0.039 39-0.787 8 μg， 0.040 72-0.814 4 μg， 0.040 45-0.809 0 μg， 0.038 63-0.772 6 μg （all r≥0.999 3）， respectively. The limits of detection were 0.008， 0.007， 0.005， 0.005， 0.020 and 0.018 μg/mL. The limits of quantitation were 0.025， 0.022， 0.015， 0.015， 0.060， 0.054 μg/mL. RSDs of precision， reproducibility and stability tests （48 h） were all lower than 1.5％ （n＝6）. Average recoveries were 98.46％， 97.06％， 100.90％， 96.13％， 96.91％， 96.57％ （RSD＜2.0％， n＝6）. CONCLUSIONS： Established method is simple， accurate and reproducible for 6 components in Chaihuang tablets， and can be used for quality control of the tablet.

Simultaneous determination of five compounds in Ganmaoning granules by UPLC-MS/MS / 国际中医中药杂志

Objective To establish a method for the simultaneous determination of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoning granules. Methods The UPLC-MS/MS method was adopted. The separation was performed on ZOBAX SB C18 (2.1 mm×150 mm, 5μm) column with mobile phase consisted of methanol-2 mmol/L ammonium acetate aqueous solution (isocratic elution) at the flow rate of 0.2 ml/min. The column temperature was set at 35℃, and the injection volume was 2μl. The electrospray ionization source (ESI) was used. Multiple reaction monitoring (MRM) mode was adopted, using positive and negative ions scanning. The ion source temperature was 300℃, the drying gas temperature was 400℃, the drying gas flow was 20 L/min, the atomizing gas pressure was 55 psi, and the capillary voltage was 4000 V.Results The linear ranges of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoling granules were 0.20-12.80 ng (r=0.9996), 1.00-64.00 ng (r=0.9998), 0.40-25.60 ng (r=0.9996), 4.00-256.00 ng (r=0.9992), 0.20-12.8 ng (r=0.9991), which showed the linear relationship was good. The limits of detection were 0.02, 0.10, 0.04, 0.40, 0.02 ng, respectively. The limits of quantitation were 0.04, 0.20, 0.08, 0.80, 0.04 ng, separately. TheRSDs of precision, reproducibility and stability (24 h) tests were no more than 3.5％. The recovery rates were 99.3％-100.75％ (RSD=2.09％-3.17％).Conclusions The method established in this experiment is simple, rapid, sensitive and accurate. It can be used to simultaneously determine the contents of five components inGanmaoling granules, and can be used for quality control ofGanmaoninggranules.

Study on the HPLC Fingerprint of Yiqing Granules / 中国药师

Objective:To establish an HPLC fingerprint of Yiqing granules to provide reference for the effective quality control. Methods:The analysis was carried out on an analytical Agilent ZORBAX SB-C18(250 mm×4.6 mm,5 μm)column with gradient elution by acetonitrile(A)-0.2% phosphoric acid solution(adjusting pH to 3.0 with triethylamine,B)(0-5 min:10% A→20%;5-20 min:20% A→30% A;20-25 min:30% A→45% A;25-35 min:45% A;35-45 min:45% A→80% A;45-50 min:80% A)at the detection wavelength of 254 nm and the flow rate of 1.0 ml·min-1,and the column temperature was 35 ℃. Twelve batches of Yiqing granules were determined by HPLC and a common mode of fingerprint was established. Results:There were 22 common peaks in the fingerprints of the twelve batches of Yiqing granules,and seven of them were identified. The similarities of fingerprints of the twelve batches of Yiqing granules were over 0.990 suggesting good reproducibility. Conclusion:The developed method is accurate and feasible,and can be used for the quality control of Yiqing granules.

Intervention impact of Wogonin on type 1 diabetic mice and its influence on p62dok expression in liver / 中国糖尿病杂志

Objective To explore the Intervention impact of Wogonin on type 1 diabetic mice and its influence on p62dok expression in liver.Methods Fifty SPF level male C57BL/6 mice were randomly divided into 5 groups:control group (Con,n =10),diabetes group (STZ group,n =40).Diabetic model was successfully made by STZ.After that,7 were chosen as model group,and then STZ group was subdivided into three subgroups according to different doses of Wogonin:5,10,and 20 mg/kg subgroup (n =10/subgroup).Fasting blood glucose levels were measured in all the subjects.And intraperitoneal injection of glucose tolerance test (IPGTT) was used to evaluate he glucose tolerance.Serum insulin level was tested by ELISA kits.Dok1,Dok2 and AKT protein expression level in liver tissue were measured by western blot.Results Wogonin could significantly reduce FPG in STZ induced type 1 diabetic mice [(12.55 ±1.31) vs (7.24 ±0.49) vs (6.22 ± 0.69) mmol/L,P ＜ 0.05].Wogonin could improve the glucose tolerance,and restore the serum insulin levels to normal in type 1 diabetic mice induced by STZ.Wogonin could increase Dok1[(0.29±0.09) vs (0.68±0.14) vs (0.79±0.13)] and Dok2[(0.32±0.08)vs (0.61±0.07) vs (0.84±0.12)] expression levels in the liver of type 1 diabetic mice induced by STZ (P＜0.05).Serum insulin-level was significantluy higher in10 mg/kg Wogonin intervention group and 20mg/kg Wogonin intervention group than in model group (P＜0.05),indicating that serum insulin level increased after Wogonin intervention.Conclusion Wogonin could significantly alleviate hyperglycemia and hyperinsulinemia in STZ induced type 1 diabetic mice,and could stimulate the expression of Dok1 and Dok2 in the liver tissue of diabetic mice.

Effect of Wogonin on osteosarcoma cells and its mechanism based on a PI3K/Akt signaling pathway / 实用肿瘤学杂志

Objective The aims of this study were to explore effects of Wogonin on proliferation,apoptosis and the PI3K/Akt signaling pathway in human osteosarcoma MG-63 cells. Methods MG-63 cells were treated with different concentrations of Wogo-nin(0,50,100 and 200 μmol/L)for 24,48 and 72 h,MTT assay was used to determine the cell proliferation,and the rates of apoptosis of MG-63 cells were assessed by the flow cytometry;Cell scratch assay was used to detect cell migration and TEM to detect morpho-logical changes of MG-63 cells;Western blot were employed to examine the expression of COX-2,Caspase-3 and P-Akt protein in MG-63 cells. Results Wogonin significantly inhibited proliferation of MG-63 cells in a dose-or time-dependent manner,and this effect was positively correlated with drug concentration and duration of action(P<0. 05). The apoptotic rate of MG-63 cells was positively correlated to the concentrations of Wogonin. Their differences were statistically significant(P<0. 05). Wogonin also signifi-cantly inhibited the migration of MG-63 cells in a dose-dependent manner(P<0. 05). Wogonin significantly down-regulated the levels of COX-2 and p-Akt protein expression and significantly up-regulated the level of caspase-3 protein expression in MG-63 cells when compared to the negative control group(P<0. 05). Conclusion Wogonin can inhibit the proliferation of MG-63 cells and promote apoptosis,which may be related to down-regulation of PI3K/Akt signaling pathway.

Anti-tumor activity of wogonin, an extract from Scutellaria baicalensis, through regulating different signaling pathways / 中国天然药物

Wogonin is a plant flavonoid compound extracted from Scutellaria baicalensis (Huang-Qin or Chinese skullcap) and has been studied thoroughly by many researchers till date for its anti-viral, anti-oxidant, anti-cancerous and neuro-protective properties. Numerous experiments conducted in vitro and in vivo have demonstrated wogonin's excellent tumor inhibitory properties. The anti-cancer mechanism of wogonin has been ascribed to modulation of various cell signaling pathways, including serine-threonine kinase Akt (also known as protein kinase B) and AMP-activated protein kinase (AMPK) pathways, p53-dependent/independent apoptosis, and inhibition of telomerase activity. Furthermore, wogonin also decreases DNA adduct formation with a carcinogenic compound 2-Aminofluorene and inhibits growth of drug resistant malignant cells and their migration and metastasis, without any side effects. Recently, newly synthesized wogonin derivatives have been developed with impressive anti-tumor activity. This review is the succinct appraisal of the pertinent articles on the mechanisms of anti-tumor properties of wogonin. We also summarize the potential of wogonin and its derivatives used alone or as an adjunct therapy for cancer treatment. Furthermore, pharmacokinetics and side effects of wogonin and its analogues have also been discussed.

Determination of Four Flavonoids in Shenyan Siwei Granules by Quantitative Analysis of Multi-components with Single Marker / 医药导报

Objective To establish an HPLC method to measure four flavonoids ( baicalin, wogonoside,baicalein and wogonin) in shenyan siwei granules by quantitative analysis of multi-components with single marker ( QAMS ) . Methods Agilent ZORBAX SB-C18 column(4. 6 mm × 250 mm,5 μm) was used. The mobile phase was composed with methanol and 0. 4% phosphoric acid solutionat at 1. 0 mL·min-1 flow rate with gradient elution. The detection wavelength was 278 nm. Baicalin was used as the internal reference substance. The relative correction factors ( RCF) between the baicalin and the other three flavonoids were established to detect the quantitation of baicalin and calculate the quantitation of the other three constituents. The external standard method was used for quantitating the four constituents, and the method was evaluated by comparing to the quantitative results between external standard method and QAMS method. Results The results of QAMS method had no significant difference with those of external standard method. Conclusion It is feasible and accurate to control the quality of shenyan siwei granules with QAMS.

Quantitative determination of multiple characteristic components in Gegen Qinlian Pills by QAMS method / 中草药

Objective To develop a quantitative analysis multi-components by single marker (QAMS) method for the quantification of multiple characteristic components, namely, puerarin, daidzin, jatrorrhizine, palmatine, soybean glycol, wogonoside, wogonin, in Gegen Qinlian Pills (GQP) with puerarin as the internal reference. Methods Samples were analyzed by Waters ultra-high efficiency liquid chromatography (UPLC) system, equipped with a reverse phase Acquity BEH C18 chromatographic column, with the mobile phase of methanol-0.1% phosphoric acid solution at a flow rate of 0.3 mL/min. The column temperature and the detection wavelength set at 30 ℃ and 260 nm respectively. Based on puerarin as the internal reference, the relative correction factors (RCFs) with other six characteristic components were calculated, then compared with the external standard method. This result benefits to verify the accuracy and advantages of the established QAMS method. Results Under the linear range of determination, RCFs of daidzin, jatrorrhizine, palmatine, soybean glycol, wogonoside, wogonin with reference to puerarin, were 1.02, 1.07, 1.05, 1.32, 0.62, and 0.90 in GQP, respectively. And the repeatability was good in different experimental conditions. Moreover, there was no significant difference on the quantitative results of seven characteristic components, derived from between external standard method and QAMS method in the 10 batches of GQP samples. The range of seven characteristic components contents in the 10 batches of GQP samples, namely puerarin, daidzin, jatrorrhizine, palmatine, soybean glycol, wogonoside and wogonin, were 8.923-10.746 mg/g, 2.231-2.988 mg/g, 0.825-1.197 mg/g, 1.274-1.522 mg/g, 2.330-2.713 mg/g, 0.836-0.951 mg/g, and 0.901-1.092 mg/g, respectively. Conclusion In present study, a feasible, convenient and accurate QAMS method with puerarin as the internal reference was established. Therefore, it is suitable to quantify multiple characteristic components in GQP and provide a useful approach for the quality control of GQP.

Effect of wogonin on OSCC cell growth and invasion / 中国病理生理杂志

AIM:To investigate the molecular mechanism of wogonin on the growth and invasion of oral squa -mous-cell carcinoma (OSCC) cell line SCC-4.METHODS:After treatment with various doses (0, 20, 40, 60, 80 and 100 mg/L) of wogonin for the indicated time , MTT assay was used to evaluate cell viability .The cell apoptosis was detec-ted by flow cytometry with Annexin V and propidium iodide (Annexin V/PI) double staining.The cell invasion ability was analyzed by Transwell assay .The activation of Wnt/β-catenin signaling molecules was assessed by Western blot .RE-SULTS:Wogonin inhibited the viability and invasion of SCC-4 cells but promoted cell apoptosis in a dose-and time-de-pendent manner .Wogonin treatment obviously decreased the activation of β-catenin.Moreover, the expression of down-stream targets cyclin D1, matrix metalloproteinase-2 (MMP-2) and MMP-9 were obviously down-regulated, accompanied by the up-regulation of anti-apoptotic protein Bcl-2.Wnt/β-catenin activator LiCl remarkably attenuated the inhibitory effect of wogonin on the activation of Wnt/β-catenin signaling molecules .Importantly, the inhibition of cell growth and in-vasion ability by wogonin treatment was dramatically attenuated after LiCl exposure .CONCLUSION: Wogonin blocks SCC-4 cell growth and invasion mainly by regulating Wnt/β-catenin signaling , indicating that it is a potential suppressor for OSCC and may be a potential target for the development of anti-OSCC therapy .

Effect of wogonin on OSCC cell growth and invasion / 中国病理生理杂志

AIM:To investigate the molecular mechanism of wogonin on the growth and invasion of oral squa -mous-cell carcinoma (OSCC) cell line SCC-4.METHODS:After treatment with various doses (0, 20, 40, 60, 80 and 100 mg/L) of wogonin for the indicated time , MTT assay was used to evaluate cell viability .The cell apoptosis was detec-ted by flow cytometry with Annexin V and propidium iodide (Annexin V/PI) double staining.The cell invasion ability was analyzed by Transwell assay .The activation of Wnt/β-catenin signaling molecules was assessed by Western blot .RE-SULTS:Wogonin inhibited the viability and invasion of SCC-4 cells but promoted cell apoptosis in a dose-and time-de-pendent manner .Wogonin treatment obviously decreased the activation of β-catenin.Moreover, the expression of down-stream targets cyclin D1, matrix metalloproteinase-2 (MMP-2) and MMP-9 were obviously down-regulated, accompanied by the up-regulation of anti-apoptotic protein Bcl-2.Wnt/β-catenin activator LiCl remarkably attenuated the inhibitory effect of wogonin on the activation of Wnt/β-catenin signaling molecules .Importantly, the inhibition of cell growth and in-vasion ability by wogonin treatment was dramatically attenuated after LiCl exposure .CONCLUSION: Wogonin blocks SCC-4 cell growth and invasion mainly by regulating Wnt/β-catenin signaling , indicating that it is a potential suppressor for OSCC and may be a potential target for the development of anti-OSCC therapy .