Effect Of Serum Containing Si Miao Yong An Decoction On Autophagy Of RAW264.7 Macrophages

Objective:To study the effect of serum containing Si Miao Yong An decoction for the autophagy of RAM264.7 macrophages, and primarily to discussion the mechanism of action for the anti-atherosclerotic effects of Si Miao Yong An decoction.Methods:The SD rats of clean grade were intervened with megadose, middle dose and small dose to prepare the drug serum. After the MTT method to observe the influence of medicated serum on cell proliferation than selecting concn which was 20% of the drug serum with each doses to intervene the RAW264.7 macrophages those cultured in vitro. Tested the expression of the LC3II by immunofluorescence labeling method, Observed mutation of the acidic autophagic vacuole by MDC staining method, Tested the mRNA’s expression level of the autophagy’s interrelate gene which were Beclinl and mTOR by RT-PCR, Tested the expression mutation of the Beclinl, LC3, P-Akt and p-mTOR albumen.Results:Compare with normal control group and immunofluorescence analysis results showed that LC3Ⅱ projected notable potentiation in serum containing Si Miao Yong An decoction processed group, MDC dyeing showed the autophagic vacuoles were obviously increased after serum containing Si Miao Yong An decoction intervened, RT-PCR display the result that serum containing Si Miao Yong An decoction induce the Beclinl mRNA’s expression and inhibit the mTOR mRNA’s expression (P<0.05 or P<0.01), Western blot’s result showed that expression of the Beclinl and LC3 IIrose, as well as expression of the p-Akt and p-mTOR were affected that inhibited obviously after serum containing Si Miao Yong An decoction intervened (P<0.05 or P<0.01).Conclusion:Si Miao Yong An decoction could induce the autophagy of RAM264.7 macrophages, its mechanism might refer the expression raising Beclinl,LC3 Ⅱ which were related autophagy genes and inhibited of Akt/mTOR signal pathways. The effect of serum containing Si Miao Yong An decoction for the autophagy of RAM264.7 macrophages, May be one of its important action mechanisms of anti-atherosclerosisan.