摘要
目的 探讨黄芪含药血清对自然杀伤(NK)细胞的杀伤活性和NKG2D、NKG2A、细胞因子干扰素-γ(IFN-γ)表达水平的影响.方法 20只SPF级SD大鼠,随机分成黄芪低、中、高剂量组(3.1、6.2、12.4 g/kg体重)和正常对照组,灌胃后制备含药血清.用NK细胞专用培养基培养NK-92MI细胞,用1640培养基培养YAC-1细胞.将NK-92MI细胞与20％不同剂量黄芪含药血清及正常对照血清孵育48 h,即分别为正常对照组、黄芪低剂量组、黄芪中剂量组以及黄芪高剂量组.通过乳酸脱氢酶(LDH)释放测定方法检测NK细胞对靶细胞YAC-1的杀伤活性,ELISA法检测IFN-γ的水平,Western blot检测NKG2D以及NKG2A蛋白表达水平.结果 与正常对照组和黄芪低剂量组相比,黄芪中、高剂量组的NK细胞活性显著升高(P＜0.01).IFN-γ的表达在黄芪低、中、高剂量组中均升高,但不随浓度升高而增强.与正常对照组相比,黄芪低、中剂量组NKG2D蛋白表达水平明显升高(P＜0.01),而NKG2A蛋白表达水平在各组间无明显差异(P＞0.05).结论 黄芪含药血清可增强NK细胞杀伤活性、上调IFN-γ表达水平、上调NK细胞活化型受体的表达,通过调控NK细胞实现对机体的保护作用.
Abstract
Objective To investigate the effect of Astragalus membranaceus on the killing activity and ex-pression levels of NKG2D,NKG2A and cytokine interferon-γ(IFN-γ)in natural killer(NK)cells.Meth-ods A total of 20 SD rats with SPF were randomly divided into low,medium and high-dose Astragalus groups(3.1,6.2,12.4 g/kg body weight)and normal control groups.NK-92MI cells were cultured with NK cell-specific medium,and YAC-1 cells were cultured with 1640 medium.NK-92MI cells were incuba-ted with 20％different doses of Astragalus membranaceus medicated serum and normal control serum for 48 h,i.e.,normal control group,Astragalus low-dose group,Astragalus medium-dose group and Astrag-alus high-dose group,respectively.The killing activity of NK cells against YAC-1 was detected by lactate dehydrogenase(LDH)release assay,the level of IFN-γ was detected by ELISA,and the protein expres-sion levels of NKG2D and NKG2A were detected by Western blot.Results Compared with normal control group and low-dose Astragalus group,the NK cell activity in the medium and high-dose Astragalus groups was significantly increased(P＜0.01).The expression of IFN-γ was increased in the low,medium and high-dose groups of Astragalus membranaceus,but it did not increase with the increase of concentration.Compared with normal control group,the expression level of NKG2D in the low and medium-dose groups of Astragalus membranaceus was significantly increased(P＜0.01),while there was no significant differ-ence in the expression level of NKG2A protein among the groups(P＞0.05).Conclusion Astragalus membranaceus medicated serum can enhance the killing activity of NK cells,up-regulate the expression level of IFN-γ,up-regulate the expression of NK cell activating receptors and achieve a protective effect on the body by regulating NK cells.
基金项目
省部共建中亚高发病成因与防治国家重点实验室开放基金(SKL-HIDCA-2019-ZY4)
NETL
NSTL
万方数据