Content determination and quality analysis of five main components in Polygala tenuifolia from different habitats / 中草药

Objective: The contents of five main components in Polygala tenuifolia from different habitats were determined, which provided certain data support and theoretical basis for the quality evaluation system of P. tenuifolia. Methods: The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin and tenuifolin of roots from different wild samples were determined by HPLC. Then SPSS 20.0 and SIMCA 11.5 were used for difference analysis, correlation analysis, hierarchical cluster analysis (HCA), and principal component analysis (PCA). Results: The content of the five main components in wild P. tenuifolia samples from different habitats was significantly different. The 20 samples were placed into two clusters (I, II) by HCA and PCA. Cluster I comprised three samples with higher content of 3,6’-disinapoyl sucrose, polygalaxanthone III, senegenin and tenuifolin from Weinan, Xianyang in Shannxi Province, and Xinjiang in Shanxi Province, whereas cluster II contained the other 17 samples. Conclusion: The results showed that the main components of P. enuifolia from Weinan, Xianyang in Shannxi Province and Xinjiang in Shanxi Province were significantly higher than other origins, and which provided a reference for the quality control, selection of excellent germplasm and cultivation bases of P. tenuifolia.

Effects of different drying methods on active constituents of root bark and root of Polygala tenuifolia / 中草药

Objective To investigate the effects of different drying methods on composition and content of five active constituents in root bark and root of Polgala tenuifoliaroot. Methods The contents of polygalaxanthone III, 3,6’-disinapoyl sucrose, polygalacic acid, senegenin, and tenuifolin in root bark and root from different drying samples were determined by HPLC. Then the data analysis was performed by ANOVA and TOPSIS methods. Results There is a difference in the order of drying methods for bark and root of P. tenuifoliaroot. For P. tenuifoliaroot root bark, the order of different drying methods was microwave drying > 60 ℃ hot-air drying > 50 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying > 40 ℃ hot-air drying > shade drying > sun drying; For P. tenuifolia root, the different drying methods were sorted by microwave drying > 60 ℃ hot-air drying > shade drying > sun drying > 50 ℃ hot-air drying > 40 ℃ hot-air drying > 70 ℃ hot-air drying > freeze-drying. Conclusion Combined with the production practice, this study suggests that microwave drying and hot-air drying at 60 ℃ are suitable drying methods for P. tenuifoliaroot bark and root, providing a basis for the determination of drying methods for the origin processing of P. tenuifolia.

Effect and mechanism of senegenin on proliferation and differentiation of neural stem cells / 国际中医中药杂志

Objective To study the effect and mechanism of senegenin on proliferation and differentiation of neural stem cells (NSCs). Methods The primary cultured NSCs were divided into the high-dose, medium-dose, low-dose group and normal control group (NC). The complete medium containing 10, 20 and 40 μmol/L senegenin was added to senegenin low-, middle-, and high- dose groups, and the NC group was routinely cultured. After 4 days of culture, CCK8 assay was used to detect cell viability, and microscopy was performed and the number of neurospheres was counted. Western blot was used to detect the expression of Nestin, TUJ1, GSK-3β, and p-GSK-3β (Ser9), and immunofluorescence staining was used to visualized Nestin and TUJ1. Results Compared with the control group, the number of NSCs neurospheres (32.78 ± 6.30, 40.93 ± 8.34, 45.37 ± 7.96 vs. 26.48 ± 5.19) and the proliferation (127.50% ± 9.31%, 138.13% ± 6.88%, 151.25% ± 9.38% vs. 100.00% ± 5.63%) in the low-, middle- and high-doses of senegenin group significantly increased (P<0.05 or P<0.01).The expression of TUJ1(2.21 ± 0.14,3.10 ± 0.16,3.30 ± 0.15 vs.1.00 ± 0.00)in the low-,middle- and high-doses of senegenin group significantly increased (P<0.05); and the expression of Nestin (0.36 ± 0.04,0.53 ± 0.05,0.46 ± 0.05 vs.1.00 ± 0.00)significantly decreased(P<0.05).The ration of p-GSK-3β(Ser9)/GSK-3β(2.31 ± 0.17,3.41 ± 0.11,3.59 ± 0.16 vs.1.00 ± 0.00)in the low-,middle-and high-doses of senegenin group significantly increased(P<0.01).The cell number of Nestin+(50.29 ± 3.18,45.28 ± 6.23,38.72 ± 5.31 vs. 75.27 ± 6.03) in the low-, middle- and high-doses of senegenin group significantly decreased (P<0.05 or P<0.01), and the cell number of TUJ1+(32.23 ± 4.36,38.23 ± 6.01,46.23 ± 4.36 vs.20.31 ± 5.23)significantly increased (P<0.01). Conclusions The senegenin may promote the proliferation and differentiation of NSCs through the activation of Wnt pathway.

Quality research and analysis of Polygala tenuifolia with different commodity grade / 中草药

Objective To establish the main component analysis method of commodity grade Polygala tenuifolia, and to explore the correlation between the grade character of P. tenuifolia and the main chemistry. Methods P. tenuifolia of four markets were determined by high performance liquid chromatography (HPLC) to analyze the different level of medicine and explore the correlation between commodity grade and composition for P. tenuifolia. Results The main components determination of P. tenuifolia in different market grades were not completely consistent with the grade of commodity. Conclusion There are certain limitations of the commodity grading standards for P. tenuifolia in the medicinal materials market. It is necessary to establish a new grade quality standard for accurately evaluating the quality of P. tenuifolia. This study also provides some reference for the comprehensive quantitative evaluation of P. tenuifolia.

Effects of Senegenin against hypoxia/reoxygenation-induced injury in PC12 cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect and the potential mechanism of Senegenin (Sen) against injury induced by hypoxia/reoxygenation (H/R) in highly differentiated PC12 cells.</p><p><b>METHODS</b>The cultured PC12 cells were treated with H/R in the presence or absence of Sen (60 μmol/L). Four groups were included in the experiment: control group, H/R group, H/R+Sen group and Sen group. Cell viability of each group and the level of lactate dehydrogenase (LDH) in culture medium were detected for the pharmacological effect of Sen. Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate. Moreover, mitochondrial membrane potential (△Ψm), reactive oxygen species (ROS) and intracellular free calcium ([Ca(2+)]i) were measured by fluorescent staining and flow cytometry. Cleaved caspase-3 and activity of NADPH oxidase (NOX) were determined by colorimetric protease assay and enzyme linked immunosorbent assay, respectively.</p><p><b>RESULTS</b>Sen significantly elevated cell viability (P<0.05), decreased the leakage of LDH (P<0.05) and apoptosis rate (P<0.05) in H/R-injured PC12 cells. Sen maintained the value of △Ψm (P<0.05) and suppressed the activity of caspase-3 (P<0.05). Moreover, Sen reduced ROS accumulation P<0.05) and [Ca(2+)]i increment (P<0.05) by inhibiting the activity of NOX (P<0.05).</p><p><b>CONCLUSION</b>Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and [Ca(2+)]i, thereby suppressing the mitochondrial pathway of cellular apoptosis.</p>

Senegenin Protects against Ischemia-reperfusion Inj ury by Ameliorating Endoplasmic Reticulum Stress in Mice / 华中科技大学学报(医学版)

Objective To investigate the protective role of senegenin(SEN)in the mouse ischemia-reperfusion injury and its mechanism.Methods Mice were divided at random into control group(the ligature was placed but not ligated for 225 min),is-chemia-reperfusion model group(the anterior descending coronary artery remained ligated for 45 min and then reperfused for 180 min)and SEN treatment group(30 mg/kg SEN was used 10 min before reperfusion).The myocardial infarct size was meas-ured by using Evans blue-TTC staining.Fluorescence assay was employed to detect the activity of Caspase-1 2 and Caspase-3 to assess the myocardial apoptosis.Western blot was used to detect the expression of two endoplasmic reticulum stress (ERS) markers,GRP78 and CHOP,in infarcted myocardia.Results Compared with the control group,the myocardial infarct size was significantly increased,the activities of Caspase-12 and Caspase-3 were increased by 4.71 fold and 3.37 fold,respectively,and the expression levels of ERS markers GRP78 and CHOP were conspicuously elevated in the ischemia-reperfusion model group.Pretreatment with SEN(30 mg/kg)could significantly reduce the myocardial infarct size,the activities of Caspase-12 and Caspase-3 and the expression levels of GRP78 and CHOP when compared with those in the ischemia-reperfusion model group and there was significant difference between the two groups(P<0.05 or P<0.01).Conclusion SEN can protect against the myocardial ischemia-reperfusion inj ury by ameliorating ERS-mediated myocardial apoptosis in mice.

Preliminary mechanism of senegenin against H/R-induced apoptosis of primary cortical neurons / 中国病理生理杂志

AIM:To explore the preliminary mechanism of senegenin ( Sen) on inhibiting hypoxia/reoxygenation ( H/R)-induced apoptosis of primary cortical neurons .METHODS:The cultured cortical neurons were randomly divided in-to normal group (control group), model group (H/R group), Sen+H/R group and Sen group.Flow cytometry was used to evaluate the effect of Sen on H/R-induced cell apoptosis .The protein levels of JNK , p-JNK, c-Jun, p-c-Jun, Bcl-2 and Bax were assessed by Western blotting .RESULTS:The apoptotic rate in H/R group was obviously higher than that in control group (P<0.05), while the apoptotic rate in Sen +H/R group was obviously lower than that in H/R group (P<0.05), suggesting that the model of apoptosis was established successfully .The results of Western blotting showed that Sen increased the expression of JNK and c-Jun, inhibited the phosphorylation of JNK and c-Jun (P<0.05), increased the protein level of Bcl-2 and inhibited the protein level of Bax in H/R treated primary cortical neurons (P<0.05).CONCLUSION:Sen has a protective effect against H/R-induced neuronal apoptosis by increasing the expression of JNK and c-Jun, inhibiting the phosphorylation of JNK and c-Jun, increasing the protein level of Bcl-2 and decreasing the protein level of Bax .

Optimal extraction of effective constituents from Radix Polygalae based on central composite design/response surface methodology / 中成药

AIM: To optimize the process of extracting effective constituents from Radix Polygalae by central composite design/response surface methodology. METHODS: Independent variables were ethanol concentration,reflux time and solvent fold,dependent variable was extraction rate of senegenin in Radix Polygalae.Linear or nolinear mathematic models were used to estimate the relationship between independent and dependent variables.Response surface methodology was used to optimize the process of extraction.Prediction was carried out through comparing the observed and predicted values. RESULTS: Regression coefficients of binomial fitting complex model was as high as 0.979 0, the optimum conditions of extraction process were 60% ethanol,2.5 hours for reflux,10-fold solvent and 2 times for extraction.Bias between observed and predicted values was-5.93%. CONCLUSION: It shows that the optimum model is highly predictive.