摘要
目的:探讨肠复方及拆方对肠癌髓源抑制性细胞(MDSCs)诱导型一氧化氮合酶(i NOS)及精氨酸酶1(ARG1)表达的影响。方法:采用不同浓度的肠复方干预肠癌CT26细胞,运用MTT法检测增殖情况,并探索肠复方最佳浓度及时间。建立小鼠结肠癌皮下移植瘤模型,21 d后取荷瘤小鼠脾脏,免疫磁珠法提取和流式细胞术检测MDSCs细胞。结合MTT试验筛选出抑制效果最佳的肠复方浓度,将20只大鼠,随机分为肠复方全方组、香菇多糖组、模型组、拆方Ⅰ组(健脾益气扶正)及拆方Ⅱ组(祛湿化瘀解毒),每组4只,制备含药血清,分组干预MDSCs细胞。运用Western blotting和RT-PCR法检测5组MDSCs细胞i NOS及ARG1的表达。结果:MTT检测显示肠复方全方对CT26细胞增殖有明显的抑制作用,且含药血清干预72 h较干预24、48 h增殖抑制作用更强,与时间呈正相关(P<0.05),肠复方中剂量组在不同干预时间段对细胞增殖的抑制作用均明显强于高、低剂量组(P<0.05)。通过流式细胞术检测小鼠脾脏MDSCs细胞为活性90%,纯度86.65%,Real-time PCR结果显示:与模型组比较,肠复方全方组、拆方Ⅰ组、拆方Ⅱ组MDSCs细胞i NOS mRNA、ARG1 mRNA表达均降低(P<0.05),以肠复方全方组最优(P<0.01),香菇多糖组有下降趋势,但差异无统计学意义(P>0.05);肠复方全方对MDSCs细胞i NOS mRNA、ARG1 mRNA抑制作用明显优于拆方Ⅰ组(P<0.05),与拆方Ⅱ组无明显差异(P>0.05)。Western blotting结果显示:与模型组比较,肠复方全方组、拆方Ⅱ组MDSCs细胞i NOS、ARG1蛋白表达均降低(P<0.05),拆方Ⅰ组、香菇多糖组差异无统计学意义(P>0.05);肠复方全方对MDSCs细胞i NOS、ARG1蛋白表达的抑制作用与拆方Ⅱ组无明显差异(P>0.05)。结论:肠复方全方及拆方能够抑制MDSCs细胞i NOS及ARG1的表达水平,肠复方全方抑制作用最优,且肠复方抗癌作用主要来自祛湿化瘀解毒组分,这可能是肠复方抑制肠癌细胞增殖和免�
Objective: To investigate the effect of Changfufang and disassembled formula on the expression of inducible nitric oxide synthase(iNOS) and arginase 1(ARG1) in myeloid-derived suppressor cells(MDSCs) of intestinal cancer. Methods: Different concentrations of Changfufang were used to intervene in the intestinal cancer CT26 cells, the proliferation was detected by the MTT method, and the optimal concentration and time of Changfufang were explored. A subcutaneous transplanted tumor model of colon cancer in mice was established.After 21 days, the spleens of tumor-bearing mice were collected, and the MDSCs cells were detected by im-munomagnetic bead extraction and flow cytometry. At the same time, combined with the MTT test, the concentration of Changfufang with the best inhibitory effect was screened, and 20 rats were randomly divided into Changfufang whole prescription group, lentinan group, model group, disassembled I group(Jianpi Yiqi Fuzheng)and disassembled II group(Qushi Huayu Jiedu), with 4 rats in each group. The drug-containing serum was prepared and the MDSCs cells were intervened in groups. The expression of iNOS and ARG1 in 5 groups of MDSCs cells was detected by Western blotting and RT-qPCR. Results: MTT test showed that Changfufang whole prescription had a significant inhibitory effect on the proliferation of CT26 cells, and the inhibitory effect of drug-containing serum on cell proliferation in 72 h was stronger than that in 24 and 48 h, which was positively correlated with time(P<0.05). The inhibitory effect of Changfufang medium-dose group on cell proliferation in different intervention time was significantly stronger than that in high-dose and low dose group(P<0.05). The activity and purity of MDSCs cells in spleen of mice were 90% and 86.65% by flow cytometry. Real-time PCR results showed that, compared with the model group, the expressions of iNOS m RNA and ARG1 m RNA in MDSCs cells of Changfufang whole prescription group, disassembled I group, and disassembled II group were all reduced(P<0.05), a
作者
邓湘琴
林宏远
崔超宇
李云辉
梁慧
DENG Xiang-qin;LIN Hong-yuan;CUI Chao-yu;LI Yun-hui;LIANG Hui(Hunan University of Traditional Chinese Medicine,Changsha Hunan 410208,China;Xiangya Hospital of Central South University,Changsha Hunan 410008,China;Hunan Cancer Hospital,Changsha Hunan 410013,China)
出处
《中医药导报》
2020年第11期33-37,共5页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
湖南省中医药管理局重点项目(201807)
湖南省自然科学基金面上项目(2017JJ2175)
长沙市科技局项目(kp1701044)。
