Analysis Of Trace Elements In Polygonum Hydropiperand/Indigofera Pseudotinctioria Mats In Guizhou And Study On The Effect Of Indigofera Pseudotinctoria Mats On Saccharomyces Cerevisiae

Objective: To study the content of metal elements in Hydropiper and Indigofera Pseudotinctioria Mats in different months,to provide experimental basis for its quality control and development and utilization,to study the effect of Indigofera Pseudotinctioria Mats Extract on Promoting Proliferation of Saccharomyces cerevisiae INVSC1 and explore the preliminary mechanisms.Methods: Determined by inductively coupled emission spectroscopy(ICP-OES)(K),magnesium(Mg),zinc(Zn),tin(Sn),copper(Cu),mercury(Hg),arsenic(As),cadmium(Cd),barium(Ba)(Al),manganese(Mn),molybdenum(Mo),antimony(Sb),cobalt(Co),gallium(Ga),germanium(Ge),lead(Pb),lithium(Ni)(Fe))and other 22 kinds of metal elements for content determination.The preparation of extract from Indigofera Pseudotinctioria Mats with 0.5、1、2、4 、 8 、 16mg/mL,Colorimetric assay(OD600)was used to detect the effects of different concentrations of extract of Indigofera Pseudotinctioria Mats on cell proliferation after 3,6,9,12,24 h Saccharomyces cerevisiae INVSC1 cells were intervened.According to the results of colorimetric method,the Indigofera Pseudotinctioria Mats(4mg/mL,8mg/mL,16mg/mL)of low、medium、high-dose and control group were set;Observation of Morphological effects of Different Concentrations of Indigofera Pseudotinctioria Mats on Saccharomyces Cerevisiae INVSC1 under Inverted Microscopy;Quantitative detection of cAMP,PKA,Tpk2 mRNA expression in Saccharomyces cerevisiae INVSC1 using real-time fluorescence quantitative PCR;Detection of cAMP,PKA expression in Saccharomyces cerevisiae INVSC1 by Western Blot.Results: The results showed that the contents of Ca,Mg,Al,K,Fe,Mn,Ag and Zn were high in Indigofera Pseudotinctioria Mats and Polygonum hydropiper,The RSD of the precision test was from 0.244% to 2.681%,and the recovery rate was from 89.9% to 107.0%.The contents of harmful heavy metal elements such as Pb,As,Cu,Hg and Cd in Polygonum Hydropiper and Indigofera Pseudotinctioria Mats are not exceeded,which is in line with the quality controlstandard of "Green Industry Standard for Import and Export of Medicinal Plants and Preparations";The colorimetric method(OD600)detected that the Indigofera Pseudotinctioria Mats had a certain proliferative effect on Saccharomyces cerevisiae INVSC1;Invertedmicroscope observation of Saccharomyces cerevisiae cells after treatment with Indigofera Pseudotinctioria Mats;Real-time fluorescence quantitative PCR detected that the medium and high doses of Indigofera Pseudotinctioria Mats could increase cAMP,PKA mRNA after 9 h of treatment;medium doses of Indigofera Pseudotinctioria Mats could increase Tpk2 mRNA;Compared with the control group,the difference was statistically significant(P<0.05);Protein cAMP,PKA expression was up-regulated by Western Blot,Compared with the control group,the difference was statistically significant(P<0.05).Conclusion:1.The content of Pb,As,Cu,Hg,Cd in Polygonum hydropiper and Indigofera Pseudotinctioria Mats was in accordance with the standard in different months.2.Different concentrations of Indigofera Pseudotinctioria Mats can promote the proliferation of Saccharomyces cerevisiae INVSC1 and change the cell morphology.3.Indigofera Pseudotinctioria Mats can increased the expression of cAMP,PKA and Tpk2 mRNA in Saccharomyces cerevisiae INVSC1.4.Extract of Indigofera Pseudotinctioria Mats can increased the expression of cAMP,PKA protein.