Mechanism of antiplatelet aggregation of active fraction from sorghum roots / 中国药理学通报

Aim To study the mechanism of anti-plate- let aggregation of sorghum root active parts. Methods The effects of active fraction (WEAE-M 30%) from sorghum roots on platelet aggregation induced by collagen, thrombin and adenosine diphosphate were investigated in vitro. Western blot, enzyme-linked immunoas-say, flow cytometry and fluorescence techniques were used to explore the mechanism of the antiplatelet aggregation effect of WEAE-M 30% . Results WEAE-M 30% had a significant inhibitory effect on platelet aggregation induced by the three agonists mentioned above. The inhibitory effect on platelet aggregation induced by collagen was the most significant, with an inhibitory rate of (72. 91 ±2. 42)%. It was found that WEAE-M 30% had a significant inhibitory effect on the collagen- mediated platelet (IPVI signaling pathway protein Src, MAPK signaling pathway protein p38 and ERK phosphorylation. It also significantly inhibited the levels of ATP, P-selection and Ca2+ in platelets. Conclusions It is suggested that the mechanism of WE-AE-M 30% antiplatelet aggregation may be related to the inhibition of platelet activation pathway GPV1, MAPK and the release of typical platelet representative particles.

Optimization of SRAP-PCR System for Valeriana officinalis var. latifolia and Primer Screening / 中国实验方剂学杂志

Objective:To establish the sequence-related amplified polymorphism （SRAP）-polymerase chain reaction （PCR） system for <italic>Valeriana officinalis</italic> var. <italic>latifolia</italic>，so as to lay the theoretical and technical foundations for the breeding of<italic> V. officinalis </italic>var. <italic>latifolia</italic>. Method:Single factor test was applied to investigate the effects of <italic>Taq</italic> Mix dose，Mg²⁺concentration，template DNA concentration，and <italic>Taq </italic>DNA polymerase content on SRAP-PCR amplification of <italic>V. officinalis </italic>var. <italic>latifolia</italic>，based on which the orthogonal experiments were performed to optimize the SRAP-PCR system for <italic>V. officinalis </italic>var. <italic>latifolia</italic>. The effective primers that could be used for genetic diversity studies of <italic>V. officinalis</italic> var. <italic>latifolia </italic>were selected under the optimal reaction condition. Result:The results of the single factor test showed that <italic>Taq </italic>Mix dose within the range of 8-11 μL resulted in better amplification. The addition of a low concentration of Mg²⁺，the medium to low concentrations of template DNA，or the low concentration of <italic>Taq</italic> DNA polymerase enhanced the amplification efficiency or richness. As demonstrated by the orthogonal experiments，the influencing degrees of related factors on SRAP-PCR amplification of <italic>V. officinalis</italic> var. <italic>latifolia </italic>were sorted in a descending order as follows： <italic>Taq</italic> Mix dose><italic>Taq</italic> DNA polymerase content>Mg²⁺ concentration>template DNA concentration. The optimal reaction system for <italic>V. officinalis</italic> var. <italic>latifolia </italic>was determined to consist of 11 μL of <italic>Taq</italic> Mix，30 ng of template DNA，0.025 mmol·L^-1Mg²⁺，1.5 U<italic> </italic>of<italic> Taq </italic>DNA polymerase，5 μmol·L^-1 forward primer，and 5 μmol·L^-1 reverse primer，which was supplemented to 20 μL with ddH₂O. The optimal annealing temperature was 36.8 ℃. A total of 17 pairs of effective primers with high band resolution and polymorphism were selected from 88 primer pairs for SRAP-PCR of <italic>V. officinalis</italic> var. <italic>latifolia</italic>. Conclusion:The established SRAP-PCR system for <italic>V. officinalis</italic> var. <italic>latifolia</italic> is stable， which can be used for genetic diversity studies of <italic>V. officinalis</italic> var. <italic>latifolia</italic>.