国际妇产科学杂志

国际妇产科学杂志 ›› 2016, Vol. 43 ›› Issue (3): 348-352.

• 论著 • 上一篇    下一篇

川芎嗪对宫颈癌HeLa细胞免疫抑制作用的影响

张雯珂，陈亚琼   

  1. 300162 天津，武警后勤学院附属医院妇产科
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2016-06-22 发布日期:2016-06-22

The Immunosuppression of Cervical Carcinoma Cell Line HeLa after Inoculating by LHC

ZHANG Wen-ke，CHEN Ya-qiong   

  1. Department of Obstetrics and Gynecology，The Affiliated Hospital of the Chinese People's Armed Police Force Logistics College, Tianjin 300162, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2016-06-22 Online:2016-06-22

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摘要： 目的：研究川芎嗪（LHC）对宫颈癌HeLa细胞免疫抑制的影响。方法：以LHC作用HeLa细胞后，培养48 h，离心后收集上清（LHC-S1），重悬，再次培养48 h，离心后收集上清（LHC-S2），重悬，再次培养48 h，离心后收集上清（LHC-S3）。以与LHC组相同条件培养未经药物作用的HeLa细胞作为对照，同样连续收集3次的培养上清（Control-S1、Control-S2、Control-S3）。用酶联免疫吸附试验（ELISA）法检测上清中转化生长因子β1（TGF-β1）、白细胞介素10（IL-10）的含量，紫外分光光度法检测前列腺素E2（PGE2）含量，噻唑蓝（MTT）法检测对植物血凝素（PHA）诱导的外周血单核细胞（PBMC）增殖及对细胞毒性T淋巴细胞（CTL）细胞毒作用的影响，以流式细胞术（FCM）分析经及不经药物作用的HeLa上清对外周血淋巴细胞亚群表型的影响。结果：（1）对照组3次培养上清中TGF-β1、IL-10和PGE2 A278 nm值差异均无统计学意义（P＞0.05）；LHC组3次培养上清中差异均有统计学意义（P＜0.05），TGF-β1逐渐降低，IL-10前2次相似，均低于第3次，PGE2 A278 nm在第2次最低；除第2次的IL-10外，LHC组IL-10均低于相应对照组。（2）3次培养上清对PBMC增殖的抑制率和对CTL的杀伤率在对照组，以及对CTL的杀伤率在LHC组和LHC组与相应对照组间差异均无统计学意义（P＞0.05）；LHC组第1次培养上清对PBMC增殖的抑制率最低。（3）对照组3次培养上清、LHC组3次培养上清外周血淋巴细胞亚群表型之间差异均有统计学意义；与对照组相比，相应LHC组的前2次培养上清CD3+、CD4+、CD25+、CD4+CD25-升高（P＜0.05），CD4+CD25+降低，第3次之间差异无统计学意义（P＞0.05）。结论：LHC可下调HeLa细胞分泌免疫抑制分子，减少免疫抑制分子对PHA诱导的PBMC增殖的抑制，降低CD4+CD25+表达，上调CD3+、CD4+、CD25+、CD4+CD25-，可在一定程度上逆转HeLa细胞的肿瘤免疫抑制。

关键词: 川芎嗪, HeLa细胞, 免疫耐受, 川芎嗪, HeLa细胞, 免疫耐受

Abstract: Ｏｂｊｅｃｔｉｖｅ：To study the variation of immunosuppression of HeLa inoculated by LHC. Methods：HeLa was inoculated by LHC，for 48 h，the supernatants were collected（LHC-S1），after centrifuging the cellｓ，and then adjusted the concentration of HeLa，inoculated for 48 h，centrifuged cells, collect the supernatants（ＬＨＣ－Ｓ２），adjusted the concentration of HeLa，inoculated for 48 h, after centrifuging the cells collected the supernatants（LHC-S3）. Compared with the HeLa′s supernatant without LHC, which was inoculated in the same condition, collected the supernatant for 3 times（Control -S1, Control -S2, Control -S3）, test the TGF-β1 and IL-10 by ELISA, PGE2 by ultraviolet spectrophotometry, by means of MTT to detect the proliferation of PHA induce PBMC and antibody-dependent cellular cytotoxicity of CTL. Flow cytometry （ＦＣＭ） ｔo analyze the subpopulation of peripheral blood lymphocytes, which was inoculated by the supernatants. Results：（１）The TGF-β1，IL-10 and PGE2 A278 nm of control group had no significant difference in statisticｓ（Ｐ＞０．０５）；ｉｎ LHC-group the difference were statically significant(P＜0.05); TGF-β1 gradually decreased, IL-10 was similarity in LHC-S1 and LHC-S２，ｌess than that in LHC-S3, PGE2 A278 nm was the minimum in LHC-S２；except in LHC-S2，IL-10 was less than the control grouｐ. （2）All the supernatants of the control group could inhibit the proliferation of PHA induced PBMC and cytotoxicity of CTL, had no significant statistic difference（P＞0.05）, the inhibition to CTL in LHC group and corresponding control group had no significant statistic difference（P＞0.05）; in LHC group, the inhibition ratio in LHC-S1 was the lest. （3）All the supernatants could influence the expression of the subpopulation of lymphocyte, it had statistic difference（P＜0.05）; Compared with control group, the percentage of CD3+，CD4+，CD25+ and CD4+CD25- increased in LHC-S1 and LHC-S2 group, the percentage of CD4+CD25+ decreased（P＜0.05), in the LHC-S3 group，ｔhe difference was not significant（P＞0.05）. Conclusions：The immunosuppressive molecules of HeLa could be reduced by LHC; LHC could decrease the inhibition of the proliferation of PHA induce PBMC; LHC also could increase the percentage of CD3+，CD4+，CD25+ and CD4+CD25-, reduce CD4+CD25+; LHC could reverse the immunosuppression of HeLa to a certain degree.

Key words: Tetramethylpyrazine, HeLa cells, Immune tolerance, Tetramethylpyrazine, HeLa cells, Immune tolerance