Studies On Flavonoids Components Analysis And Against Proliferation Of Human Hepatocellular From Scutellaria Barbata D.Don

Scuteuaria barata D.Don is a perennial herb of the Lamiaceae.The main physiological active substances are flavonoids.The origin of Scuteuaria barata D.Don is wide and the chemical composition in these different regions still exists individual differences.In this study,the methods for the determination of the total flavonoids,nine flavonoid compounds and eight microelements in Scuteuaria barata D.Don from ten places was researched,which could be adopetd for multi-component quality control of Scuteuaria barata D.Don.The clinical research shows that Scuteuaria barata D.Don has been widely used for treating liver diseases,such as liver cancer and hepatitis.This study takes HepG-2 as research subjects to explore the the inhibitory effect of nine flavonoids on HepG-2 proliferation by MTT assay method,inverted microscope which was used to observe cell morphological change and flow cytometer which analyzed the distribution of cell cycle.In addition,the apoptosis mechanism of HepG-2 cell induced by Scutellarin was discussed.This dissertation consists of five parts.1.The contents of total flavonoids were measured by UV-VIS spectrophotometry The total flavonoids of Scuteuaria barata D.Don from ten different places were determined by UV-VIS spectrophotometry,with Rutin as reference substance.The total flavonoids contents of Wenzhou,Nanyang,Suzhou,Baoding,Gutian,Bozhou,Huanggang,Taiyuan,Meizhou and Yulin is 8.67%,5.78%,5.72%,4.42%,4.21%,3.78%,3.75%,3.32%,1.78%and 1.12%,respectively.2.Simultaneous determination of nine kinds of flavonoids in Scutellaria barbata D.Don by HPLC An HPLC method for simultaneous determination of 9 flavonoids monomer of Scutellaria barbata D.Don was established.The ingredients of flavonoids are various from different places,and the range of contents are as follows:Scutellari:0.492%?0.713%,Baicalein:0.021%?0.04%,Apigenin:0.041%?0.141%,Quercetin:0.015%?0.271%,Luteolin:0.011%?0.236%,Wogonin:0.027%?0.087%,Baicalin:0.006%?0.034%,Naringenin:0.006%?0.023%,Scutellarein:0.032%?0.087%.3.Determination of eight microelements in Scutellaria barbata D.Don by high-temperature digestion-ICP-OES The inductively coupled plasma atomic emission spectrometer was adopted to determine the contents of Mn,Cd,Cr,Cu,Fe,Ni,Pb,Zn with the samples dissolved by concentrated nitric acids and hydrogen peroxide.It is found that the content of Fe was highest and the range of content was 868.38?5046.45 mg/Kg.The second was Mn,with 138.13?453.79 mg/Kg.And the contents of the eight metallic elements from ten places were different.4.To screen activity flavonoids compounds Against Proliferation of Human Hepatocellular from Scutellaria barbata D.Don The inhibitive rate of HepG-2 growth by nine flavonoids compounds of Scuteuaria barata D.Don was investigated.The MTT results showed that the growth inhibitory effect of HepG-2 cells induced by flavonoids was time-dependent and dose-dependent.After the cells exposed to HepG-2 for 48h,the inhibiting effect was Scutellarin>Quercetin>Luteolin>Apigenin>Baicalin>Baicalein>Wogonin.Naringenin and Scutellarein was not obvious.5.The mechanism of Apoptosis of HepG-2 induced by Scutellarin was researched The MTT assay was used to examine the effect of Scutellarin on the proliferation of HepG-2 cell,morphology observation and flow cytometry was adopted for cell cycle distribution.The results showed that Scutellarin could induce apoptosis in the S phase,and it was concentration-dependent.the cell population of the G2/M phase was reduced concurrently with the increase of the S phase population.Theantitumor activity based on HepG-2 was implemented through apoptosis,which might be related to the cell arrest in S phase.