Functional Mechanism On Aralia Echinocaulis Inhibit RA Synovial Membrane Proliferation Based On PI3K/Akt/mTOR Signal Pathway

Objective: To observe the effect of Aralia Echinocaulis on proliferation of RA fibroblasts based on the PI3K/Akt/mTOR signaling pathway and to explore its possible mechanism of action on rheumatoid arthritis.Methods: The MH7 A cell line was selected as the experimental research object.Firstly,the effect of Aralia echinocaulis alcohol extracts on the proliferation of MH7 Acells was determined by CCK-8 method,and the appropriate drug concentration wasselected.The MH7 A cells were treated with Aralia echinocaulis alcohol extracts with a variety of concentration(0?g/ml?550?g/ml?650?g/ml?750?g/ml)working on the effect of inducing MH7 A cells apoptosis at 24 h,and then to detect their apoptosis by Annexin V-FITC/PI double staining;MH7A cells were divided into blank group,TNF-? group,low-dose group,medium-dose group,and high-dose group.Except the blank group,the other groups were stimulated with LPS(1?g/ml)for 6h.According to the CCK-8 test results,the drug group was treated with different concentrations of Aralia echinocaulis alcohol extracts(550?g/ml ? 650?g/ml ? 750?g/ml),and the cells in each group were further cultured for 24 h.The levels of inflammatory factors(TNF-?,IL-1?)in the supernatant of each group were detected by ELISA.The expression of PI3K?P-Akt and Becline1 protein in each group was detected by Western-blot.Results: After the MH7 A cells were treated with the Aralia echinocaulis alcohol extracts,the inhibition rate increased significantly with the increase of administration concentration(P<0.05);Aralia echinocaulis alcohol extracts with different concentrations(0 ? 550 ? 650 ? 750?g/ml)stimulates MH7 A cells for 24 h,apoptosis rate of 24 h are 2.20%?3.07%?5.27%?8.04%;When MH7 A cells were induced by LPS,the expression of IL-1? and TNF-? in supernatant was significantly up-regulated compared with normal group(P<0.05);Compared with the model group,the expression of the two groups was down-regulated with different concentration of drug stimulation(P<0.05);After 6 hours of LPS stimulation,compared with the normal group,the expression of PI3 K and P-Akt in the model group increased(P<0.05),and the expression of Becline1 decreased(P<0.05);Compared with the model group,the expression of PI3 K and P-Akt decreased in groups of the drug group(P<0.05),and Becline1 increased(P<0.05).Conclusion: The Aralia echinocaulis alcohol extracts can inhibit the proliferation of MH7 A cells and significantly promote the apoptosis of MH7 A cells,inhibit the expression of IL-1? and TNF-? in MH7 A cells,down regulate the protein expression of PI3 K and p-Akt,and up regulate the protein expression level of becline1.Therefore,the Aralia echinocaulis alcohol extracts may promote autophagy and inhibit the expression of inflammatory factors by inhibiting the PI3 K/Akt/mTOR signal pathway.