Effects Of Radix Astragali Extract And Boswellin Extract On The Proliferation Of NIH3T3 Cell

For studying the effects of Radix Astragali Extract(RAE)and Boswellin Extract(BE)on cell proliferation,the S phase change of cell cycle,the mRNA of EZH1/EZH2 and UTX/JMJD3 and ERK/p-ERK.Cultured cells in vitro,Cell batches,Randomly divided into two groups were treated respectively with RAE and ME in different concentration.24h later,the cell growth effects of RAE and BE was detected by using CCK-8 method.The cell cycle of NIH3T3 cell was detected by using FCM.The mRNA of EZH1/EZH2 and UTX/JMJD3 was detected by RT-PCR.The ERK/p-ERK was detected by Western Blotting.1.The CCK-8 results showed that there was extremely significant difference with RAE concentration of 8×10-3g/L,1.6×10-3g/L,3.2×10-4g/L and BE concentration of 4×10-1 and 8×10-2g/L to interven cells for 24h(P<0.01).2.The S phase change of cell cycle using FCM also proves with the RAE concentration of 8×10-3g/L,1.6×10-3g/L,3.2×10-4g/L and BE concentration of 4×10-1g/L,8×10-2g/L and 1.6×10-2g/L,S phase proportion was extremely significant difference(P<0.01).3.The results of RT-PCR showed that different concentration of RAE and BE on the expression EZH1 mRNA of NIH3T3,RAE concentration of 8×10-3g/L,1.6×10-3g/L,3.2×10-4g/L and BE concentration of 4×10-1g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).The expression EZH2 mRNA of NIH3T3,RAE concentration of 8×10-3g/L and 1.6×10'3g/L and BE concentration of 4×10-1g/L and 8×10-2g/L.RAE concentration of 3.2×10-4g/L,there was extremely significant difference(P<0.01).4.The expression UTX mRNA,RAE concentration of 3.2×10-4g/L and BE concentration of 8×10-2g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).The expression JMJD3 mRNA,RAE concentration of 8×10-3g/L and 3.2×10-4g/L and BE concentration of 8×10-2g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).5.After the NIH3T3 was treated with U0126,the results of RT-PCR showed that different concentration of RAE and BE on the expression EZH1 mRNA of NIH3T3,RAE concentration of 8×10-3g/L,1.6×0-3g/L,3.2×10-4g/L and BE concentration of 4×10-1g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).The expression EZH2 mRNA of NIH3T3,RAE concentration of 8×10-3g/L and 1.6×10-3g/L and BE concentration of 4×10-1g/L,8×10-2g/L and 1.6×10-2g/L,there was a statistical significance comparing with the control group respectively(P<0.01).6.The expression UTX mRNA,RAE concentration of 3.2×10-4g/L and BE concentration of 4×10-1g/L,8×10-2g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).The expression JMJD3 mRNA,RAE concentration of 1.6×10-3g/L and 3.2×10-4g/L and BE concentration of 4×10-1g/L,8×10-2g/L and 1.6×10-2g/L,there was extremely significant difference(P<0.01).7.After treated cell with drug 24h,there was no difference with ERK1/2.p-ERK1/2 increased with the drug concentration increased,in a dose-dependent manner.After the NIH3T3 was treated with U0126,there was no difference with ERK1/2.p-ERK1/2 increased with the drug concentration increased,in a dose-dependent manner.