不同类别抗瘤中药制剂下调结直肠癌免疫功能抑制的比较研究

目的比较研究不同类型抗瘤中药制剂对结直肠癌NK、T细胞免疫功能抑制的影响。方法分别制备三氧化二砷（As2O3）、川芎嗪（LHC）、黄芪（AMB）、氧化苦参碱（MOX）、猪苓多糖（PUPS）、蒿甲醚（ART）等6种中药制剂作用后的结直肠癌细胞Colon26再培养上清,以不经中药制剂作用的相应上清为对照;MTT法检测不同中药制剂作用后对Colon26所致小鼠脾细胞NK杀伤活性、ConA诱导转化抑制的影响,直接免疫荧光FCM法检测对小鼠脾细胞IL-2Rα、CD3ε＋ζ＋及CD3ε-ζ＋表达抑制的影响。结果As2O3、PUPS及ART对NK杀伤抑制的下调约50%左右,LHC及AMB可下调约75%左右;AMB的下调作用最强。LHC和PUPS作用后对CD3ε-ζ＋表达抑制的初始下调幅度约80%~90%,AMB和ART作用后初始下调幅度约40%~50%,MOX可完全消除抑制并明显促进表达;LHC的下调作用最强。AMB、PUPS及ART对T细胞诱导转化抑制的下调程度较强（下调约50%）,维持时间较长;ART对IL-2Rα表达抑制的下调作用最强（下调约50%）,维持时间较长;AMB对CD3ε＋ζ＋表达抑制的下调作用最强,不但可完全消除表达抑制,还可进一步促进表达,维持时间亦较持久。结论不同类型抗瘤中药制剂下调结直肠癌免疫抑制的类别、程度、出现和维持的时间不尽相同;下调肿瘤细胞产生的NK、T细胞免疫功能抑制是中药制剂抗瘤效应机制之一。

Comparision of down-regulating effects of different anti-tumor traditional Chinese medicines on immunosuppressions caused by colorectal tumor cells

Objective To study the down-regulating effects of different anti-tumor traditional Chinese medicines on inhibition of NK and T cell immunofunctions caused by colorectal tumor cells.Methods Re-cultured supernatants of Colon26（colorectal tumor cells） pre-treated by six different traditional Chinese medicines,including As2O3,Ligustrazine Hydrochloride（LHC）,Astragalus Mongholicus Bge（AMB）,Matrine N-Oxide（MOX）,Polyporus Umbellatus Polysacchride（PUPS） and Artemether（ART）,and the corresponding supernatants of Colon26 treated without any medicines as control were collected.The effect on inhibition of NK killing activity and Con-A-induced transformation of murine splenocytes were measured by MTT,and that on inhibition of IL-2Rα,CD3ε＋ζ＋ and CD3ε-ζ＋ expressions were detected by FCM of direct immunofluorescence labeling.Results The decreasing levels of inhibited NK killing activity caused by As2O3,PUPS and ART were about 50%,and those caused by LHC and AMB were about 75%;the decreasing effect of AMB was strongest.The initial decreasing levels of inhibited CD3ε-ζ＋ expression caused by LHC and PUPS were about 80%-90%,those caused by AMB and ART were about 40%-50%,and that was reversed completely caused by MOX and elevated expression of CD3ε-ζ＋ expression was significant;the decreasing effect of LHC was strongest.The levels of down-regulating effect of AMB,PUPS and ART on inhibited transformation of T cell were stronger（about 50%） with the longer duration time;the down-regulating effect of ART on inhibited IL-2Rα expression of T cell was strongest（about 50%） with longer duration time;the down-regulating effect of AMB on inhibited CD3ε＋ζ＋ expression of T cell was strongest,which was reversed completely and the elevated expression was significant.Conclusions The down-regulating effects of different anti-tumor traditional Chinese medicines on colorectal tumor immunosuppressions are distinguished among sorts,levels,occurring and keeping times.Reducing the immunosuppressions of NK and T cell caused by tumor cells should be one of new anti-tumor mechanisms of traditional Chinese medicines.