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DOI:
:2017,(5):16-21
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慢性再生障碍性贫血患者中医证型与外周血microRNA、淋巴细胞亚群及细胞因子的关系研究
李峻1,陈劼2,孙雪梅1,朱学军1,姜鹏君1,陈健一1,倪海雯1,张文曦1,夏雯1
(1.南京中医药大学附属医院(江苏省中医院)血液科(江苏南京210029) ;2.南京中医药大学附属医院(江苏省中医院)病理科(江苏南京210029))
Study on Correlation between TCM Syndromes and Peripheral Blood MicroRNA,Lymphocyte Subsets and Cytokines in Patients with Chronic Aplastic Anemia
LI Jun1, CHEN Jie2, SUN Xue-mei1, ZHU Xue-jun1, JIANG Peng-jun1, CHEN Jian-yi1, NI Hai-wen1, ZHANG Wen-xi1, XIA Wen1
(1.Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine (Jiangsu Province Hospital of Traditional Chinese Medicine), Nanjing, Jiangsu, 210029;2.Department of Pathology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine (Jiangsu Province Hospital of Traditional Chinese Medicine), Nanjing, Jiangsu, 210029)
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投稿时间:2017-05-09    
中文摘要:   目的:分析慢性再生障碍性贫血(CAA)患者外周血microRNA、淋巴细胞亚群、细胞因子水平与不同肾虚证型的关系。  方法:纳入60例CAA患者,辨证分为肾阴虚组30例、肾阳虚组14例、肾阴阳两虚组16例,另选择20例健康志愿者作为正常对照组。采用实时荧光定量PCR方法检测受试者外周血单个核细胞miRNA-155-5p、miRNA-1260b表达情况,采用流式细胞术检测外周血淋巴细胞亚群包括总T淋巴细胞(T)、辅助T淋巴细胞(Th)、细胞毒T淋巴细胞(Tc)、B淋巴细胞(B)、自然杀伤细胞(NK),采用酶联免疫吸附法检测血浆干扰素-γ(IFN-γ)、白介素-12(IL-12)p70、肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)水平。  结果:与正常对照组比较,肾阴虚组、肾阴阳两虚组、肾阳虚组患者的miRNA-155-5p、miRNA-1260b表达及Tc比例、IFN-γ、IL-12 p70、TNF-α水平均明显升高(P<0.01,P<0.05),NK细胞比例、Th比例、Th/Tc比值及TGF-β1水平均明显降低(P<0.05,P<0.01)。与肾阳虚组比较,肾阴虚组、肾阴阳两虚组患者的miRNA-155-5p、miRNA-1260b表达及Tc比例、IFN-γ、IL-12 p70、TNF-α水平均明显升高(P<0.01,P<0.05),Th比例、Th/Tc比值及TGF-β1水平均明显降低(P<0.05,P<0.01);肾阴虚组患者的Th比例低于肾阴阳两虚组(P<0.05),血浆IFN-γ、IL-12 p70、TNF-α水平高于肾阴阳两虚组(P<0.05)。  结论:肾阴虚型、肾阴阳两虚型较肾阳虚型患者存在更严重的细胞免疫功能亢进及miRNA-155-5p、miRNA-1260b表达失调,而肾阴虚型较肾阴阳两虚型患者存在更严重的造血负调控因子紊乱。
Abstract:  Objective:To analyze the relationship between the levels of peripheral blood microRNA, lymphocyte subsets and cytokines and different types of kidney deficiency syndrome in patients with chronic aplastic anemia (CAA).   Methods: A total of 60 patients with CAA were included, who were divided into kidney yin deficiency group (30 cases), kidney yang deficiency group (14 cases) and kidney yin and yang deficiency group (16 cases) by syndrome differentiation, meanwhile another 20 cases of healthy volunteers were included as normal control group. The expressions of miRNA-155-5p and miRNA-1260b in mononuclear cells of peripheral blood were detected by real-time quantitative PCR. The proportion of peripheral blood lymphocyte subsets including total T lymphocyte (T), helper T lymphocyte (Th), cytotoxic T lymphocyte (Tc), B lymphocyte (B) and nature killer cell (NK) were measured by flow cytometry. The plasma levels of IFN-γ, IL-12 p70, TNF-α and TGF-β1 were determined by enzyme-linked immunosorbent assay.   Results: Compared with the normal control group, the expressions of miRNA-155-5p and miRNA-1260b, the proportion of Tc and the levels of IFN-γ, IL-12p70 and TNF-α were obviously increased in the kidney yin deficiency group, kidney yang deficiency group and kidney yin and yang deficiency group (P<0.01,P<0.05), and the proportions of NK and Th, the Th/Tc ratio and the level of TGF-β1 were significantly decreased (P<0.05,P<0.01). Compared with the kidney yang deficiency group, the expressions of miRNA-155-5p and miRNA-1260b, the proportion of Tc and the levels of IFN-γ, IL-12p70 and TNF-α were obviously increased in the kidney yin deficiency group and kidney yin and yang deficiency group (P<0.01,P<0.05), and the proportion of Th, the Th/Tc ratio and the level of TGF-β1 were markedly decreased (P<0.05,P<0.01). The proportion of Th in the kidney yin deficiency group was lower than that in the kidney yin and yang deficiency group (P<0.05), and the levels of IFN-γ, IL-12p70 and TNF-α were higher than those in the kidney yin and yang deficiency group (P<0.05).   Conclusion: Compared with the patients of kidney yang deficiency type, the patients of kidney yin deficiency type and kidney yin and yang deficiency type show more serious cellular immune hyperfunction and dysregulation of miRNA-155-5p and miRNA-1260b, and compared with the patients of kidney yin and yang deficiency type, the patients of kidney yin deficiency type show more serious disorder of hematopoietic negative regulatory factors.
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基金项目:江苏省自然科学基金资助项目(BK20131034);江苏省政府留学奖学金资助项目苏教外(2015)44号;江苏省卫生国际(地区)交流支撑计划青年医师自选项目苏卫国合(2016)155号
Author NameAffiliation
LI Jun Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
CHEN Jie Department of Pathology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
SUN Xue-mei Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
ZHU Xue-jun Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
JIANG Peng-jun Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
CHEN Jian-yi Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
NI Hai-wen Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
ZHANG Wen-xi Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
XIA Wen Department of Hematology, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, 210029 
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