| Objective Observing the influence of the Flavonoids from Tetrastigma hemsleyanum Diels et Gilg on the expression of Regulatory T Cells(Treg)in vivo and related cytokines in vitro of Lewis Lung Cancer Mice.Preliminary study the Possible mechanisms of Tetrastigma hemsleyanum Diels et Gilg flavone reverse the immune escape of tumor by Regulating immune function of the tumor mice,to provide experimental evidence for Clinical medicine.Methods①lewis Lung Cancer Mice models were established by inoculated subcutaneously way,and were randomly divided into normal group,NS group,Celecoxib group,Tetrastigma hemsleyanum Diels et Gilg flavone high-dose group,middle-dose group,low-dose group.24h after inoculation,the treatment groups were began to intragastric administration one time every day,the normal group was not treated.②Administration for 14 and 23days,half of each group were sacrificed animal,separated of mononuclear cells from spleen,the expression of Treg cell proportion and CTLA-4 were measured by flow cytometry.③lewis Lung Mice models were established by inoculated subcutaneously way,14 days after inoculation,Separated and cultured the spleen mononuclear cell,by TGF-β1 stimulation,with Celecoxib group as positive control,use the ELISA method to observe Flavonoids from Tetrastigma hemsleyanum Diels et Gilg on spleen mononuclear cells secreting PGE2,COX-2 limitation,dose effect relationship.④One-way ANOVA and Repeated Measure were used for statistical analysis of the corresponding samples.Results ①Tumor volume.Compared with the NS group,the Celecoxib group,high-dose and middle-dose group were significantly decreased,with a very difference in tumor volume on D23(P<0.01).Within the groups,there were comparative differences(P<0.05).Tumors size in the order:high-dose group<middle-dose group<low-dose group.Celecoxib group was close to middle-dose group(P>0.05).②Treg ratio in the mononuclear cells of spleen:Compared with the normal group,all model groups were significantly increased,with a very significantdifference in ratio of Treg cells(P<0.01).Compared with the NS group,the treatment groups were significantly decreased,with a very difference in ratio of Treg cells on D23(P<0.01).Within the groups,there were comparative differences(P<0.01).Treg ratio in the order:high-dose group<middle-dose group<low-dose group.Celecoxib group was close to middle-dose group(P>0.05).③CTLA-4(CD152)ratio in Treg cells of Spleen mononuclear:Compared with the normal group,all model groups were significantly increased,with a very significant difference in CD152 ratio of Treg cells(P<0.01).Compared with the NS group,the treatment groups were significantly decreased,with a difference in CD152 ratio of Treg cells on D23(P<0.05).The Celecoxib group was the best one in downregulating the expression of CD152,compared with middle-dose group and low-dose group,there was a significant differences(P<0.05).There was no significant differences between the rest groups(P>0.05).④Spleen mononuclear cells secrete PGE2 and COX-2 content in vitro:The results show that both the PGE2 and COX-2 content were increased firstly and then decreased,while the highest was at 24H.Compared with the blank control group,the remaining 5 groups were significantly increased,with a very significant difference in content of PGE2 and COX-2(P<0.01).Compared with the model group,the 4 drug groups were significantly decreased,with a very difference in content of PGE2 and COX-2(P<0.01).Within the groups,there were comparative differences(P<0.05).The content of PGE2 and COX-2 in the order:Celecoxib group<high-dose group<middle-dose group<low-dose groupConclusion①There was a positive correlation between the effect of tumor inhibition and dose of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg.The hig-dose group of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg had most remarkable effect on inhibiting tumor.Celecoxib group was close to middle-dose group.②Tumor inhibitory effect of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg may be associated with reducing Treg ratio in mononuclear cells,which had a positive correlation with the dose of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg,The first was high-dose group,Celecoxib group was close to middle-dose group.③Tumor inhibitory effect of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg may be associated with reducing CD152 ratio in Treg cells.But there was no significant difference between each dose group.On D23,Celecoxib group had the best effect,which was close to the high-dose group,and superior to middle-dose group and low-dose group.④Flavonoids from Tetrastigma hemsleyanum Diels et Gilg may affect PGE2,COX-2 expression in vitro,to improve tumor immune microenvironment and anti tumor,which had a positive correlation with the dose of Flavonoids from Tetrastigma hemsleyanum Diels et Gilg.At 24H,Celecoxib group had the best effect,high-dose group was superior to middle-dose group. |
PDF Full Text Request