艾灸对胃荷瘤大鼠外周血中CD4~+、CD4~+CD25~+T淋巴细胞的影响
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摘要
目的:观察艾灸对胃荷瘤大鼠外周血中CD4~+、CD4~+CD25~+T淋巴细胞亚群的影响。方法:将50只健康SPF级SD大鼠,雌雄各半,随机分为空白组、假手术组、模型组、艾灸组和红外组5组,每组10只。利用Walker-256细胞来源的实体瘤组织移植胃部制备模型。模型成功后,艾灸组予以悬灸中脘、胃俞等穴位;红外组予以短波红外线照射腹部胃脘及背部胃脘投影区;其余组捆绑束缚。干预20 min/次,1次/d,持续21 d。干预结束后,眼眶采血,流式细胞技术测定外周血中CD4~+、CD4~+CD25~+T淋巴比例。处死动物,测算胸腺指数及肿瘤体积生长抑制率。结果:与空白组比较,模型组动物生存状态变差,胸腺指数增加,外周血中CD4~+CD25~+T淋巴细胞比例显著增加(P <0. 05);与模型组比较,艾灸组动物艾灸组动物生存状态改善,胸腺指数进一步增加,瘤体内坏死灶较多,血管较少,外周血中CD4~+T淋巴细胞比例显著增加(P <0. 01),CD4~+CD25~+T淋巴细胞比例减少(P <0. 01);与红外组比较,艾灸组瘤体内坏死灶更多,血管较少,外周血中CD4~+T淋巴细胞比例增加(P <0. 01)。结论:艾灸可一定程度改善荷瘤动物生存状态,减少肿瘤细胞转移、促进肿瘤细胞坏死,减少血管增生。这种影响可能与增加CD4~+T淋巴细胞,减少CD4~+CD25~+T淋巴细胞有关。
Objective: To observe the effects of moxibustion on CD4~+and CD4~+CD25~+T lymphocytes in peripheral blood of rats with gastric cancer. Methods: A total of 50 healthy SPF SD rats( half male and half female) were randomly divided into 5 groups:the blank group,the sham operation group,the model group,the moxibustion group and the infrared group,with 10 rats in each group. Solid tumor tissue derived from Walker-256 cells was transplanted into the stomach to prepare the tumor model. After modeling,moxibustion group and infrared group were treated 20 mins per day for 21 days. After the intervention,the blood from the orbits was collected. And the ratio of CD4~+and CD4~+CD25~+T lymphocytes in peripheral blood was tested by flow cytometry. After killing the rats,the thymus index and the growth inhibition rate of the tumor volume were measured. Results: Compared with the blank group,the survival status of the model group was worse,the thymus index increased,and the proportion of CD4~+CD25~+T lymphocytes in peripheral blood significantly increased( P < 0. 05). Compared with the model group,the survival status of the moxibustion group was improved,the thymus index was further increased,more necrosis and less blood vessels were found in the tumors,and the percentage of CD4~+T lymphocytes was increased significantly( P < 0. 01),while that of CD4~+CD25~+T lymphocyte was decreased( P < 0. 01). Compared with the infrared group,more necrosis and few blood vessels were found in the tumors,and the proportion of CD4~+T lymphocytes was increased in the moxibustion group( P < 0. 01). Conclusion: Moxibustion can improve the survival status of the rat with gastric tumor,inhibit tumor metastasis,promote tumor necrosis as well as prevent vessels hyperplasia. These changes may be related to the increasing CD4~+T lymphocytes and the decreasing CD4~+CD25~+T lymphocytes.
Objective: To observe the effects of moxibustion on CD4~+and CD4~+CD25~+T lymphocytes in peripheral blood of rats with gastric cancer. Methods: A total of 50 healthy SPF SD rats( half male and half female) were randomly divided into 5 groups:the blank group,the sham operation group,the model group,the moxibustion group and the infrared group,with 10 rats in each group. Solid tumor tissue derived from Walker-256 cells was transplanted into the stomach to prepare the tumor model. After modeling,moxibustion group and infrared group were treated 20 mins per day for 21 days. After the intervention,the blood from the orbits was collected. And the ratio of CD4~+and CD4~+CD25~+T lymphocytes in peripheral blood was tested by flow cytometry. After killing the rats,the thymus index and the growth inhibition rate of the tumor volume were measured. Results: Compared with the blank group,the survival status of the model group was worse,the thymus index increased,and the proportion of CD4~+CD25~+T lymphocytes in peripheral blood significantly increased( P < 0. 05). Compared with the model group,the survival status of the moxibustion group was improved,the thymus index was further increased,more necrosis and less blood vessels were found in the tumors,and the percentage of CD4~+T lymphocytes was increased significantly( P < 0. 01),while that of CD4~+CD25~+T lymphocyte was decreased( P < 0. 01). Compared with the infrared group,more necrosis and few blood vessels were found in the tumors,and the proportion of CD4~+T lymphocytes was increased in the moxibustion group( P < 0. 01). Conclusion: Moxibustion can improve the survival status of the rat with gastric tumor,inhibit tumor metastasis,promote tumor necrosis as well as prevent vessels hyperplasia. These changes may be related to the increasing CD4~+T lymphocytes and the decreasing CD4~+CD25~+T lymphocytes.
引文
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[6]易受乡,彭艳,常小荣,等.艾灸对应激性胃溃疡大鼠胃黏膜细胞增殖和凋亡的影响及其与热休克蛋白表达关系的研究[J].针刺研究,2006,31(5):259-263+271+256.
[7]易受乡,郁洁,常小荣,等.艾灸及槲皮素预处理对胃黏膜细胞凋亡及HSP70表达的影响[J].中华中医药学刊,2009,27(11):2245-2248.
[8]欧阳里知,谭静,赵欢,等.艾灸对荷瘤胃癌大鼠外周血中CD3-CD161+NK与CD3+CD161+NKT含量的影响[J].湖南中医药大学学报,2018,38(5):541-545.
[9]隋英忠,夏玉军,王晓静,等.构建SD大鼠移植性胃癌模型及不同方法诱癌率的比较[J].中国临床研究,2014,27(2):135-137.
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[11]Zhao P,Dai M,Chen W,et al.Cancer trends in China[J].Japanese Journal of Clinical Oncology,2010,40(4):281-285.
[11]George M.Hahn.Hyperthermia and Cancer[M].New York:New York Plenum Press,1982:2-3.
[12]李鼎九.肿瘤热疗学[M].2版.郑州:郑州大学出版社,2003:1-2.
[13]Benjamin Frey,Eva-Maria Weiss,ect.Old and new facts about hyperthermia-induced modulations of the immune system[J].2012,28(6):528-542.
[14]张贺龙,刘文超.临床肿瘤学[M].西安:第四军医大学出版社,2016:156-160.
[15]黄金昶.发挥针灸治疗肿瘤优势促迚中西医结合发展[J].中国针灸,2018,38(10):1123-1124.
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[17]郭义.实验针灸学[M].北京:中国中医药出版社,2016:127-130.
[18]惠鑫,黄畅,王昊,等.艾烟在艾灸中的作用机制及安全性[J].世界中医药,2017,12(9):2246-2251.
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[20]张雅丽,侯安丽,王杏茶,等.CD4+CD25+Foxp3+调节性T细胞在子宫内膜癌患者外周血中的表达及意义[J].现代免疫学,2015,35(5):410-414.
[21]Barao I,Hanash A M,Hallett W,et al.Suppression of Natural Killer Cell-Mediated Bone Marrow Cell Rejection by CD4+CD25+Regulatory T Cells[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(14):5460-5465.
[22]Chen X,Du Y,Lin X Q,et al.CD4+CD25+regulatory T cells in tumor immunity[J].International Immunopharmacology,2016,34(2):244-249.
[23]徐林,徐薇,蒋正刚,等.CD4+CD25+CCR6+调节性T细胞在小鼠乳腺癌模型中对CD8+T细胞的抑制作用[J].中国肿瘤生物治疗杂志,2009,16(5):431-435.
[24]Gao Y L,Chai Y F,Qi A L,et al.Neuropilin-1high CD4+CD25+Regulatory T Cells Exhibit Primary Negative Immunoregulation in Sepsis[J].Mediators of Inflammation,2016,2016(9):1-11.
[25]Wang L Y,Zeng Y,Pan Z Z,et al.Detection of intracellular and extracellular cytokines of CD4+CD25+regulatory T cells in gastric cancer patients][J].Chinese Journal of Cancer,2007,26(3):270.
[26]Chen C,Chen D,Zhang Y,et al.Changes of CD4+CD25+FOXP3+and CD8+CD28-regulatory T cells in non-small cell lung cancer patients undergoing surgery[J].International Immunopharmacology,2014,18(2):255-261.
[27]马金环.急性胃出血患者早期血液CD4+CD25+T细胞及其细胞因子的变化[J].中国实验诊断学,2017,21(7):1170-1172.
[28]龚单春,张海东,于振坤,等.CD4~+CD25~+T regs与CCL17和CCL22在头颈鳞状细胞癌发病机制中的初步研究[J].临床耳鼻咽喉头颈外科杂志,2017,31(20):1557-1560.
[29]施怡,李晓斌,章岳山,等.结肠癌患者化疗/热化疗前后CD4+CD25+T细胞和CTL变化[J].中国癌症杂志,2010,20(8):626-629.
[30]Hou P F,Zhu L J,Chen X Y,et al.Age-related changes in CD4+CD25+FOXP3+regulatory T cells and their relationship with lung cancer:[J].Plos One,2017,12(3):e0173048.
[2]Global,regional,and national under-5 mortality,adult mortality,agespecific mortality,and life expectancy,1970-2016:a systematic analysis for the Global Burden of Disease Study 2016[J].The Lancet,2017,390(10100):1084-1150.
[3]中日医学科技交流协会热疗专业委员会.中国肿瘤热疗临床应用指南(2017.V1.1)[J].中华放射肿瘤学杂志,2017,26(4):369-375.
[4]Isomoto H,Ohtsuru A,Braiden V,et al.Heat-directed suicide gene therapy mediated by heat shock protein promoter for gastric cancer[J].Oncology Reports,2006,15(3):629-35.
[5]彭艳,易受乡,封迎帅,等.艾灸对幽门螺杆菌胃炎大鼠血清免疫学作用研究[J].中国针灸,2014,34(8):783-790.
[6]易受乡,彭艳,常小荣,等.艾灸对应激性胃溃疡大鼠胃黏膜细胞增殖和凋亡的影响及其与热休克蛋白表达关系的研究[J].针刺研究,2006,31(5):259-263+271+256.
[7]易受乡,郁洁,常小荣,等.艾灸及槲皮素预处理对胃黏膜细胞凋亡及HSP70表达的影响[J].中华中医药学刊,2009,27(11):2245-2248.
[8]欧阳里知,谭静,赵欢,等.艾灸对荷瘤胃癌大鼠外周血中CD3-CD161+NK与CD3+CD161+NKT含量的影响[J].湖南中医药大学学报,2018,38(5):541-545.
[9]隋英忠,夏玉军,王晓静,等.构建SD大鼠移植性胃癌模型及不同方法诱癌率的比较[J].中国临床研究,2014,27(2):135-137.
[10]严洁,朱兵.针灸的基础与临床[M].长沙:湖南科技出版社,2010:549-647.
[11]Zhao P,Dai M,Chen W,et al.Cancer trends in China[J].Japanese Journal of Clinical Oncology,2010,40(4):281-285.
[11]George M.Hahn.Hyperthermia and Cancer[M].New York:New York Plenum Press,1982:2-3.
[12]李鼎九.肿瘤热疗学[M].2版.郑州:郑州大学出版社,2003:1-2.
[13]Benjamin Frey,Eva-Maria Weiss,ect.Old and new facts about hyperthermia-induced modulations of the immune system[J].2012,28(6):528-542.
[14]张贺龙,刘文超.临床肿瘤学[M].西安:第四军医大学出版社,2016:156-160.
[15]黄金昶.发挥针灸治疗肿瘤优势促迚中西医结合发展[J].中国针灸,2018,38(10):1123-1124.
[16]侯新芳,倪光夏.艾灸治疗恶性肿瘤研究进展[J].世界中西医结合杂志,2014,9(1):101-104.
[17]郭义.实验针灸学[M].北京:中国中医药出版社,2016:127-130.
[18]惠鑫,黄畅,王昊,等.艾烟在艾灸中的作用机制及安全性[J].世界中医药,2017,12(9):2246-2251.
[19]朱彤波.医学免疫学[M].2版.四川:四川大学出版社,2017:275.
[20]张雅丽,侯安丽,王杏茶,等.CD4+CD25+Foxp3+调节性T细胞在子宫内膜癌患者外周血中的表达及意义[J].现代免疫学,2015,35(5):410-414.
[21]Barao I,Hanash A M,Hallett W,et al.Suppression of Natural Killer Cell-Mediated Bone Marrow Cell Rejection by CD4+CD25+Regulatory T Cells[J].Proceedings of the National Academy of Sciences of the United States of America,2006,103(14):5460-5465.
[22]Chen X,Du Y,Lin X Q,et al.CD4+CD25+regulatory T cells in tumor immunity[J].International Immunopharmacology,2016,34(2):244-249.
[23]徐林,徐薇,蒋正刚,等.CD4+CD25+CCR6+调节性T细胞在小鼠乳腺癌模型中对CD8+T细胞的抑制作用[J].中国肿瘤生物治疗杂志,2009,16(5):431-435.
[24]Gao Y L,Chai Y F,Qi A L,et al.Neuropilin-1high CD4+CD25+Regulatory T Cells Exhibit Primary Negative Immunoregulation in Sepsis[J].Mediators of Inflammation,2016,2016(9):1-11.
[25]Wang L Y,Zeng Y,Pan Z Z,et al.Detection of intracellular and extracellular cytokines of CD4+CD25+regulatory T cells in gastric cancer patients][J].Chinese Journal of Cancer,2007,26(3):270.
[26]Chen C,Chen D,Zhang Y,et al.Changes of CD4+CD25+FOXP3+and CD8+CD28-regulatory T cells in non-small cell lung cancer patients undergoing surgery[J].International Immunopharmacology,2014,18(2):255-261.
[27]马金环.急性胃出血患者早期血液CD4+CD25+T细胞及其细胞因子的变化[J].中国实验诊断学,2017,21(7):1170-1172.
[28]龚单春,张海东,于振坤,等.CD4~+CD25~+T regs与CCL17和CCL22在头颈鳞状细胞癌发病机制中的初步研究[J].临床耳鼻咽喉头颈外科杂志,2017,31(20):1557-1560.
[29]施怡,李晓斌,章岳山,等.结肠癌患者化疗/热化疗前后CD4+CD25+T细胞和CTL变化[J].中国癌症杂志,2010,20(8):626-629.
[30]Hou P F,Zhu L J,Chen X Y,et al.Age-related changes in CD4+CD25+FOXP3+regulatory T cells and their relationship with lung cancer:[J].Plos One,2017,12(3):e0173048.