免疫增效方对结肠癌Lovo细胞免疫耐受微环境的调节作用  

Modulatory effects of immunopotentiation decoction on the immune tolerance microenvironment of colon cancer cell Lovo

作　　者：崔宇[1,2] 庞雁[2] 吴晓静[2] 牛雪姣[1] 姚嫱[1,2] 胡人杰[3] 

机构地区：[1]天津市中医药大学,天津300193 [2]天津市人民医院,天津300121 [3]天津市医药科学研究所,天津300020

出　　处：《生物医学工程与临床》2014年第3期275-278,共4页Biomedical Engineering and Clinical Medicine

基　　金：天津市卫生局中西医结合科研课题(07038)

摘　　要：目的研究免疫增效方水提物对结肠癌Lovo细胞株免疫耐受微环境的调节作用。方法选择由黄芪（40g）、女贞子（20g）、当归（15g）、枸杞子（15g）、白术（10g）、五味子（10g）组成的免疫增效方，利用高速离心联合乙醇提取法制备水提物；利用四甲基偶氮唑盐（MTT）法检测自2．34～300．00μg/μL8个浓度梯度免疫增效方水提物培养液作用24、48h后L0vo细胞的增殖抑制率；利用酶联免疫吸附分析（ELISA）法检测高（150．00μg/μL）、中（37．50μg/μL）、低（4．69μg/μL）药物浓度作用后细胞培养液上清液转化生长因子β1（TGF—β1）、白细胞介素10（IL-10）水平的变化。结果与对照组相比，不同浓度免疫增效方均可不同程度抑制细胞的增殖，且呈剂量及时间依赖性，作用24h及48h的IC50分别为34．80μg/μL和16．61μg/μL；TGF-β1水平随药物浓度升高而减少，同样作用时间不同质量浓度组间差异具有统计学意义（24h：P〈0．05；48h：P〈0．01）：而IL-10水平随药物浓度升高而增加，同样作用时间不同质量浓度组间差异具有统计学意义（24h：P〈0．05；48h：P〈0．01）。结论免疫增效方水提物可明显抑制Lovo细胞增殖，具有-定的抗肿瘤活性．且可抑制TGF-β1产生．拮抗肿瘤相关的免疫耐受，此外随剂量增加，IL-10表达呈上升趋势，而IL-10可通过促进CD8＋T细胞的增殖和分化，发挥免疫促进作用，以上的结果均有助于机体免疫系统对肿瘤的杀伤．并能-定程度提高抗肿瘤免疫治疗效果。Objective To investigate the effect of immunopotentiation decoction on the immune tolerance microenvironment of colon cancer cell line Lovo. Methods The immunopotentiation decoction constituted of Astragalus membranaceus（40 g）, Ligustrum lucidum（20 g）, Angelica（15 g）, Lycium barbarum（15 g）, Rhizome attractylodes（10 g）, and Schisandra chinensis（10 g） were prepared and extracted by high speed centrifugation and ethanol. The methyl thiazoly tetrazolium salt （MTT） method was used to detect the viability of Lovo cells 24-hour or 48-hour after incubation with 8 different concentrations of immunopotentiation decoction water extract ranging from 2.343 75 to 300.000 00 μg/μL. Enzyme-linked immnnosorbent assay（ELISA） method was used to detect the level of transforming growth factor（TGF- β1） and interleukin-10（IL-10） in the cell culture supernatant after incubation with high concentration（150.00 μg/μL）, middle concentration（37.50 μg/μL） and low concentration（4.69 μg/μL） of the drug extract. Results Compared with the control group, immunopotentiation decoction extract demonstrated an inhibitory effect on the cell proliferation in a dose and time dependent manner, with IC50 of 24-hour and 48-hour incubation were 34.80μg/μL and 16.61μg/μL respectively. The level of TGF- β1 decreased with increasing drug concentration, and significant differences were observed among different concentration groups at the each time point（24-hour. P 〈 0.05; 48-hour. P 〈 0.01）. The level of IL-10 increased with increasing drug concentration, and the differences among the different concentration groups at the each time point were statistically significant（24- hour. P 〈 0.05; 48-hour. P 〈 0.01）. Conclusion It is demonstrated that the immunopotentiation decoction water extract could inhibit the proliferation of Lovo cells, which has certain antitumor activity. The water extract could also inhibit the secretion of TGF- β1, thereby antagonizing the tumor-associated immune

关 键 词：结直肠癌 免疫增效方 免疫耐受 转化生长因子β1(TGF—β1) 白细胞介素10(IL-10) 

分 类 号：R735.35[医药卫生—肿瘤]