Study On The Function And The Immune Mechanism Of Against Hepatic Carcinoma Of Aiv, β-elemene

AIM:1.Through observing the influence of AIV andβ-elemene to immunologic function of Mφand DC in vitro, and observing if it has synergism of AIV andβ-elemene,we will discuss whether AIV andβ-elemene can enhance the immunologic function of Mφand DC.2.Through observing the influence of AIV andβ-elemene to cytokines such as IL-2 IFN-γ,TNF-a in vitro, we will discuss whether AIV andβ-elemene can enhance the immunologic function of the mice,and whether it has synergism of AIV andβ-elemene.3. Through observing the inhibition of AIV and P-elemene to the subcutaneous tumor of mice with liver tumor, and detecting T lymphocytes and IFN-γ,IL-4 in splenocytes, we will discuss the influence of AIV andβ-elemene to the immune system of mice with liver tumor and the antitumor effect.METHODS:Study in vitro:Cultivate mice macrophages J447A.1 in vitro. Four groups are taken during logarithmic growth phase, Control group, AIV group,β-elemene group, unity group.Detected its surface molecules--MHCⅡ,MHCⅠ,CD40,CD86 with FCM. Detected the content of TNF-a,IL-1βin cell supernatant with ELISA. Detected the content of NO in cell supernatant with Griess. Cultivate mice DCs D2SC in vitro. Five groups are taken during logarithmic growth phase, Control group, LPS group, AIV group,β-elemene group, unity group. Detected its surface molecules-- MHC II,MHC I,CD40,CD80,CD86 with FCM. We detected the content of IL-6,IL-12in cell supernatant with ELISA. Study in vivo:The C57 mice were randomly divided into AIV group, P-elemene group, unity group and control group.Give the mice of control group with NS, give the mice of AIV group with AIV, give the mice of P-elemene group withβ-elemene, give the mice of unity group with AIV and P-elemene.The drug concentration is 100μg/ml every mouse. Collect blood from retrobulbar in 7 days,14 days,21days after administration. Detected the content of IL-2,IFN-γ,TNF-a in serum with ELISA. Established subcutaneous tumor model of hepatocellular carcinoma. The mice were randomly divided into AIV group, P-elemene group, unity group and control group. Give the mice of control group with NS, give the mice of AIV group with AIV, give the mice of P-elemene group with P-elemene, give the mice of unity group with AIV and P-elemene.The drug concentration is 1001g/ml every mouse. Tumor growth was observed in 15 days after administration. Detected the expression of CD4,CD8 with FCM.RESULTS:Study in vitro:The expression of Mφsurface molecules-- MHCⅡ,MHC I,CD40,CD86 with the treatment of AIV andβ-elemene is significantly higher than the control group(P<0.01, P<0.05). The expression of MHCⅡ, CD40,CD86 of unity group is significantly higher than AIV group(P<0.05). The expression of MHC I of unity group is significantly higher thanβ-elemene group(p<0.05). The ability of the macrophages swallow the neutral red dye is gradually increased with the increasing of concentration and time of AIV andβ-elemene. The ability of the macrophages swallow the neutral red dye is the best after 24h, the concentration of 100μg/ml. There is a synergy when combined using AIV andβ-elemene. The expression of TNF-α,IL-1p,NO after the treatment of various concentrations of AIV andβ-elemene is more than control group. There is statistically significant when the concentration of 100μg/ml(p<0.01,p<0.05). The expression gradually increased with the increasing of concentration of AIV andβ-elemene. The expression of TNF-α,IL-1β.NO in unity group is more than the alone group. The expression of MHC II,MHC I,CD40,CD80,CD86 after the treatment of AIV and P-elemene is significantly higher than the control group,the LPS group(p<0.01, p<0.05). The expression of MHC II,MHC I,CD40,CD80,CD86 after the treatment of both is significantly higher than the single group (p<0.05). The expression of IL-12,IL-6 after the treatment of AIV andβ-elemene is significantly higher than the control group,the LPS group. The expression of IL-12 IL-6 in AIV group and P-elemene group is significantly higher than the LPS group(p<0.05). The expression of IL-12,IL-6 in unity group is significantly higher than the LPS group(p<0.01). The expression of IL-12,IL-6 in unity group is significantly higher than the single group (p<0.05). Study in vivo:The expression of IL-2,IFN-y,TNF-a of the mice after the treatment of AIV and P-elemene is significantly higher the control group (p<0.01,P<0.05). The expression is gradually increased with the increasing of time. The expression of IL-2,IFN-y,TNF-αof the mice after the 21 days treatment of AIV andβ-elemene is significantly higher the single group (p<0.05). The tendency of growth of the subcutaneous tumor of 3 experimental group is obviously slower than the NS group after the treatment of AIV and P-elemene. The subcutaneous tumor had the decrease tendency after 13 days treatment of AIV andβ-elemene.Theβ-elemene group is more obviously than the AIV group. Quantity of T cells in experimental control group and experimental group is obviously decreased than the control group. Quantity of T cells in experimental group is obviously increased than experimental control group. Quantity of T cells in unity group is obviously increased than single group. The proportion of T cell subsets is obviously changed between normal mice and tumor-bearing mice.The proportion of CD4+/CD8+ increased after the treatment of AIV and P-elemene. The proportion of the unity group is obviously increased CD4+/CD8+>1(P<0.05). The expression of IFN-y,IL-4 of tumor-bearing mice after the treatment of AIV andβ-elemene is obviously higher than the control group. The expression of IFN-y,IL-4 of AIV group andβ-elemene group is obviously higher than the control group(P<0.05). The expression of IFN-γ,IL-4 of unity group is obviously higher than the control group(P<0.01). The expression of IFN-y,IL-4 of unity group is obviously higher than single group(P<0.05).CONCLUSION:1.AIV andβ-elemene has significantly inhibitory effect on liver tumor-bearing mice. There is a synergy when combined using AIV andβ-elemene.2. AIV andβ-elemene can enhance cellular and humoral immune function with stimulating the activation of Mcp, promoting the maturation of DC, promoting high expression of immune-related cytokines.