| 三叶青提取物对体外培养NK细胞功能的影响 |
| 投稿时间：2016-11-09 修订日期：2017-06-02 点此下载全文 |
| 引用本文：王莹,章松平,俞哲,徐爱芳.三叶青提取物对体外培养NK细胞功能的影响[J].医学研究杂志,2018,47(2):155-159 |
| DOI：
10.11969/j.issn.1673-548X.2018.02.037 |
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| 中文摘要:目的 三叶青提物对体外培养NK细胞功能的影响。方法 用物理和化学方法分离和纯化三叶青中的有效成分，并用高效液相色谱（HPLC）法鉴定提取纯度。分离慢性乙肝患者外周血单个核细胞（PBMC）；在体外用磁分法从PBMC中分离NK细胞；用NK细胞培养基将分离后的NK细胞进行定向培养。收集培养第7天的NK细胞给予不同浓度的三叶青提取液诱导72h：用CCK-8法检测人NK细胞增殖率；流式细胞术（FCM）检测三叶青提取物作用前后NK细胞穿孔素（PFP）、颗粒酶B （GrB）、CD107a的表达；FCM法检测三叶青提取物诱导72h后NK细胞的IFN-γ表达。结果 将三叶青根块分别提取9个部分（分别命名为TH-t、TH-p、TH-a、TH-b、TH-w、TH-w1、TH-w2、TH-w3和TH-w4）。各部位三叶青提取物对NK细胞均有促进生长作用，差异有统计学意义（P<0.05），在药物浓度相同条件下，以水煮脱糖部位（Th-w3）时促NK细胞生长活性最强；总提取物（Th-t）、石油醚部位（Th-p）和水部位脱糖流分次之，与对照组比较差异均有统计学意义（P<0.05）。三叶青总提取物作用NK细胞72h后，其穿孔素和颗粒酶B阳性表达均明显升高，以0.62μg/ml三叶青总提取物组的穿孔素和颗粒酶B阳性表达率达到最明显，与对照组比较差异有统计学意义（P<0.05）。经三叶青提取物诱导的NK细胞其IFN-γ表达以浓度为0.62μg/ml时最高（99.01%），较对照组（88.46%）显著上升（P<0.05）。结论 三叶青提取物能促进慢性乙型肝炎患者NK细胞增殖和功能改变，上调NK细胞表面PFP、GrB、CD107a和IFN-γ的表达。 |
| 中文关键词:慢性乙型肝炎 NK细胞 三叶青/提取物 |
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| Effect of the Extract of Radix Tetrastigmatis on the Function of NK Cells in vitro |
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| Abstract:Objective To study the effect of the extract of radix tetrastigmatis on the function of NK cells in vitro.Methods The effective components of radix tetrastigmatis were separated and purified by physical and chemical methods, and the extraction purity were identified by HPLC. Peripheral blood mononuclear cells (PBMC) were separated in patients with chronic hepatitis B. NK cells were separated from PBMC by magnetic separation in vitro. The separated NK cells were in orientational culture with NK cell medium. The NK cells cultured for seven days were collected and induced for 72 hours by the extract of radix tetrastigmatis at different concentrations. Human NK cell proliferation was detected by CCK-8 assay. The expression of NK cells perforin (PFP), granzyme B (GrB) and CD107a detected by flow cytometry (FCM) before and after the effect by the extract of radix tetrastigmatis. The expression of NK cells IFN-γ was detected by flow cytometry (FCM) after 72h-induced.Results Nine parts of the radix tetrastigmatis were extracted (named TH-t, TH-p, TH-a, TH-b, TH-w, TH-w1, TH-w2, TH-w3 and TH -w4). All parts of the extract could promote the growth of NK cells, and the differences were significant. Under the same conditions of concentration, water boiling desugar portion (Th-w3) promoted the growth most actively. Total extract (Th-t), petroleum ether (Th-p) and water fractions desugared followed, and the differences were significant with the control group (P<0.05). After 72h induced by the total extract of radix tetrastigmatis on NK cells, the positive expression rate of perforin and granzyme B was significantly increased, and total extract group(0.62μg/ml) reached the highest expression, compared with the control group (P<0.05). The expression rate of IFN-γ was 99.01% with its concentration of 0.62μg/ml, which was significantly higher than the expression of control group (88.46%) (P<0.05).Conclusion The extract of radix tetrastigmatis can promote the proliferation and function changes of NK cells in patients with chronic hepatitis B, and up-regulate the NK cell surface expression of PFP, GrB, CD107a and IFN-γ. |
| keywords:Chronic Hepatitis B NK cell Radix Tetrastigmatis/Extract |
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