Enhancement of proliferation and anti-tumor activity of human peripheral blood lymphocytes by low-intensity pulsed ultrasound:a experimental study

HUANG Denggao; GAO Yuanhui; CAO Hui; ZHENG Linlin; WEN Xiaohong; FU Xianxian; ZHANG Shufang

doi:10.16245/j.cnki.issn1008-6978.2020.01.001

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Enhancement of proliferation and anti-tumor activity of human peripheral blood lymphocytes by low-intensity pulsed ultrasound:a experimental study

更新时间：2021-10-08

- Enhancement of proliferation and anti-tumor activity of human peripheral blood lymphocytes by low-intensity pulsed ultrasound:a experimental study
- Journal of Clinical Ultrasound in Medicine Vol. 22, Issue 1, Pages: 1-5(2020)
- 作者机构：
  
  中南大学湘雅医学院附属海口医院中心实验室
- 作者简介：
- 基金信息：
- DOI：10.16245/j.cnki.issn1008-6978.2020.01.001
  CLC： R734.2;R445.1
- Published：2020，
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[1]黄邓高,高元慧,曹卉,郑琳麟,文小红,符先先,张淑芳.低强度脉冲超声增强人外周血淋巴细胞增殖及抗癌活性的实验研究[J].临床超声医学杂志,2020,22(01):1-5.

HUANG Denggao, GAO Yuanhui, CAO Hui, et al. Enhancement of proliferation and anti-tumor activity of human peripheral blood lymphocytes by low-intensity pulsed ultrasound:a experimental study. [J]. Journal of Clinical Ultrasound in Medicine 22(1):1-5(2020)
[1]黄邓高,高元慧,曹卉,郑琳麟,文小红,符先先,张淑芳.低强度脉冲超声增强人外周血淋巴细胞增殖及抗癌活性的实验研究[J].临床超声医学杂志,2020,22(01):1-5. DOI： 10.16245/j.cnki.issn1008-6978.2020.01.001.

HUANG Denggao, GAO Yuanhui, CAO Hui, et al. Enhancement of proliferation and anti-tumor activity of human peripheral blood lymphocytes by low-intensity pulsed ultrasound:a experimental study. [J]. Journal of Clinical Ultrasound in Medicine 22(1):1-5(2020) DOI： 10.16245/j.cnki.issn1008-6978.2020.01.001.

摘要

目的探讨低强度脉冲超声（LIPUS）对人外周血淋巴细胞（HPBL）增殖和抗肿瘤效应的影响。方法采用Ficoll梯度离心法分离HPBL;将LIUPS强度0设为对照组

10 mW/cm

组、20 mW/cm

组、30 mW/cm

组、40 mW/cm

组、50 mW/cm

组为刺激组刺激HPBL增殖

每天一次

每次刺激10 min

连续刺激2 d。CCK-8法检测各组细胞增殖活性;酶联免疫吸附法检测抗肿瘤因子[γ干扰素（INF-γ）、白介素2（IL-2）、肿瘤坏死因子α（TNF-α）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）]表达情况;采用共培养HPBL与人肺巨细胞癌95D细胞检测其抗肿瘤活性。结果 LIPUS刺激HPBL2 d共2次后

30 mW/cm

组的细胞增殖能力增强

与对照组和其他强度刺激组比较

细胞数量和细胞团块数量均增多。CCK-8检测结果显示

30 mW/cm

组光密度值与对照组、20 mW/cm

组、40 mW/cm

组、50 mW/cm

组比较差异均有统计学意义（P=0.032、0.027、0.027、0.001）。抗肿瘤因子检测显示

30 mW/cm

组INF-γ、IL-2、TNF-α和GM-CSF分别为（39.66±8.30）ng/L、（619.84±125.94）ρg/ml、（461.29±14.08）ng/L、（108.11±2.07）ng/L

均高于对照组[（24.40±6.84）ng/L、（279.56±128.18）ρg/ml、（417.17±21.44）ng/L和（96.53±1.56）ng/L]和50 mW/cm

组[（16.52±0.87）ng/L、（141.00±93.71）ρg/ml、（171.96±9.46）ng/L、（72.56±8.95）ng/L]

差异均有统计学意义（均P

0.01）。共培养实验显示

30 mW/cm

组人肺巨细胞癌95D细胞杀伤率为51.11%±5.21%

显著高于对照组（42.86%±0.55%）和50 mW/cm

组（42.00%±3.67%）

差异均有统计学意义（均P

0.05）。结论 LIPUS刺激可提高HPBL的增殖活性和杀伤率。

Abstract

ObjectiveTo investigate the effect of low intensity pulsed ultrasound（LIPUS）on the proliferation and anti-tumor effect of human peripheral blood lymphocytes（HPBL）.MethodsFicoll gradient centrifugation was used to separate HPBL.LIUPS of 0 intensity was set as the control group

and the stimulation group was 10 mW/cm

20 mW/cm

30 mW/cm

40 mW/cm

and 50 mW/cm

respectively

then HPBL proliferation was stimulated by stimulation groups for 10 min/d for two consecutive days.The proliferation activity of the cells was measured by the CCK-8 assay.The expressions of anti-tumor factors interferon-γ（INF-γ）

interleukin 2（IL-2）

tumor necrosis factor α（TNF-α）and granulocyte-macrophage colony stimulating factor（GM-CSF） in the supernatant of culture medium were detected by the ELISA assay. The anti-tumor activity was detected by co-culture of HPBL and human lung giant cell carcinoma cells（95 D）.ResultsAter 2 d stimuli

30 mW/cm

group significantly promoted HPBL proliferation compared with the control group and other stimulation groups after stimulated for two consecutive days（P

0.01）.The results of CCK-8 showed that the optical density of 30 mW/cm

group was significantly different from that of the control group

20 mW/cm

group

40 mW/cm

group and 50 mW/cm

group

respectively（P=0.032

0.027

0.001）.Anti-tumor factor detection showed that the levels of INF-γ

IL-2

TNF-αand GM-CSF in 30 mW/cm

group were（39.66±8.30）ng/L

（619.84±125.94）ρg/ml

（461.29±14.08）ng/L

（108.11±2.07）ng/L

respectively

which were higher than those in the control group[（24.40±6.84）ng/L

（279.56±128.18）ρg/ml

（417.17±21.44）ng/L and（96.53±1.56）ng/L]and 50 mW/cm

group[（16.52±0.87）ng/L

（141.00±93.71）ρg/ml

（171.96±9.46）ng/L

（72.56±8.95）ng/L]

and the differences were statistically significant（all P

0.01）.Co-culture experiment showed that the killing rate of 95 D cells in 30 mW/cm

group was 51.11%±5.21%

whichwassignificantlyhigherthanthatinthecontrolgroup（42.86%±0.55%）and 50 mW/cm

group（42.00%±3.67%）

there were significant（both P

0.05）.ConclusionThe proliferation activity and killing rate of HPBL were increased after LIPUS stimulation.

关键词

超声低强度脉冲淋巴细胞外周血人细胞因子细胞增殖抗肿瘤

Keywords

Ultrasoundlow intensity pulsedPeripheral blood lymphocyteshumanCytokinesCell proliferationAnti-tumor

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