Danshen Injection Inhibits SKOV3 Cell Proliferation <italic style="font-style: italic">in Vitro</italic> by Interfering with Their Interaction with Platelets

Ping YUAN; Si-qin JIANG; Yi-ran NIE; Sheng-ling LIU; Wei-ran FU; Hui-ru TIAN; Jian-jiang FU

doi:10.13422/j.cnki.syfjx.20212324

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Danshen Injection Inhibits SKOV3 Cell Proliferation in Vitro by Interfering with Their Interaction with Platelets

更新时间：2021-10-29

- Danshen Injection Inhibits SKOV3 Cell Proliferation in Vitro by Interfering with Their Interaction with Platelets
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 23, Pages: 59-65(2021)
- 作者机构：
  
  1.江西中医药大学药学院，南昌 330004
  2.江西中医药大学国际教育学院，南昌 330004
  3.江西中医药大学生命科学学院，南昌 330004
  4.首都医科大学基础医学院，北京 100069
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20212324
  CLC： R22;R242;R2-031;R285.5
- Received：09 August 2021，
  
  Published Online：15 October 2021，
  
  Published：05 December 2021
- 稿件说明：
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袁萍,姜思琴,聂亦然等.丹参注射液通过干扰肿瘤细胞与血小板相互作用抑制SKOV3细胞体外增殖[J].中国实验方剂学杂志,2021,27(23):59-65.

YUAN Ping,JIANG Si-qin,NIE Yi-ran,et al.Danshen Injection Inhibits SKOV3 Cell Proliferation in Vitro by Interfering with Their Interaction with Platelets[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(23):59-65.
袁萍,姜思琴,聂亦然等.丹参注射液通过干扰肿瘤细胞与血小板相互作用抑制SKOV3细胞体外增殖[J].中国实验方剂学杂志,2021,27(23):59-65. DOI： 10.13422/j.cnki.syfjx.20212324.

YUAN Ping,JIANG Si-qin,NIE Yi-ran,et al.Danshen Injection Inhibits SKOV3 Cell Proliferation in Vitro by Interfering with Their Interaction with Platelets[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(23):59-65. DOI： 10.13422/j.cnki.syfjx.20212324.

摘要

目的

观察丹参注射液对肿瘤细胞与血小板相互作用诱导的卵巢癌细胞增殖的抑制作用。

方法

采用噻唑蓝（MTT）比色法和集落形成法，观察血小板对SKOV3体外生长的诱导作用及丹参注射液（12，24，48 g·L

-1

）的抑制作用；采用酶联免疫吸附测定法（ELISA）检测血小板-肿瘤细胞相互作用系统及血小板上清液中转化生长因子-

（TGF-

）含量，并观察丹参注射液对TGF-

分泌的影响；采用微量板法和ELISA观察肿瘤细胞培养上清液对血小板聚集和分泌的影响，并检测丹参注射液的抑制作用；采用蛋白免疫印迹法（Western blot）观察丹参注射液对血小板核转录因子-

B（NF-

B）信号转导通路的影响及单独用丹参注射液的影响。

结果

与空白组比较，血小板诱导组吸光度

570

显著升高（

0.01）；血小板＋丹参注射液组

570

则较血小板诱导组显著降低（

0.01），且与丹参注射液组比较，经血小板诱导的同剂量丹参注射液组细胞增殖抑制率更显著（

0.01）。与空白组比较，血小板诱导组集落数明显增加（

0.05）；血小板＋丹参注射液的集落数则明显低于血小板诱导组（

0.05，

0.01）。与空白组比较，血小板诱导组上清液中TGF-

含量显著升高（

0.01）；血小板＋丹参注射液组TGF-

含量显著降低（

0.01）；与丹参注射液（24 g·L

-1

）组比较，经血小板诱导的同剂量丹参注射液组抑制率更高（

0.01）。与空白组比较，经丹参注射液处理的血小板上清中TGF-

明显减少（

0.05，

0.01）；经丹参注射液处理的SKOV3上清中TGF-

的含量差异无统计学意义。与空白组比较，SKOV3细胞上清液诱导组的血小板聚集、血栓素A

（TXB

）和5-羟色胺（5-HT）分泌显著升高（

0.01），细胞上清液诱导+丹参注射液组上述指标均显著降低（

0.01），且与丹参注射液（24 g·L

-1

）组比较，经细胞上清液诱导的同剂量丹参注射液组抑制率更高（

0.01）。与空白组比较，血小板诱导组的磷酸化TGF-

激活激酶1（p-TAK1），p-NF-

B表达显著升高，p-NF-

B抑制蛋白（p-I

B）表达显著降低（

0.01）；血小板+丹参注射液组p-TAK1，p-NF-

B表达均显著降低，p-I

B表达显著升高（

0.01）。

结论

丹参注射液可通过抑制血小板与肿瘤细胞相互作用从而对SKOV3细胞增殖产生的影响，其作用机制可能与抑制血小板分泌TGF-

有关。

Abstract

Objective

To investigate the inhibitory effects of Danshen injection against ovarian cancer cell proliferation induced by the interaction between platelets and cancer cells.

Method

The induction of platelets on SKOV3 growth

in vitro

and the inhibitory effect of Danshen injection at 12，24，and 48 g·L

-1

were observed by 3-（4，5-Dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） and cell colony formation assays. The content of transforming growth factor-

（TGF-

） in the platelet-tumor cell interaction system and platelet supernatant and the effect of Danshen injection on TGF-

secretion were detected by enzyme-linked immunosorbent assay （ELISA）. The influences of tumor cell culture supernatant on platelet aggregation and secretion and the inhibitory effect of Danshen injection were determined by microplate assay and ELISA. The effects of Danshen injection on platelet nuclear factor kappa B （NF-

B） signaling pathway were assayed by Western Blot.

Result

Compared with the blank group， the platelet induction group exhibited significantly elevated absorbance at

570

（

0.01）， while the absorbance at

570

in the platelet + Danshen injection group was significantly lower than that in the platelet induction group （

0.01）. The comparison with the Danshen injection group revealed that the cell proliferation inhibitory rate in the platelet + Danshen injection group at the same dose was more significant （

0.01）. The number of colonies in the platelet induction group was obviously increased in contrast to that in the blank group（

0.05）， while the number of colonies in the platelet + Danshen injection group was significantly lower than that in the platelet induction group（

0.05，

0.01）. As demonstrated by comparison with the blank group， TGF-

content in the supernatant of the platelet induction group rose remarkably（

0.01）， whereas that in the platelet + Danshen injection group declined（

0.01）. Compared with the Danshen injection （24 g·L

-1

） group， the platelet + Danshen injection group displayed more obvious inhibition（

0.01）. Compared with the blank group， Danshen injection significantly reduced the TGF-

content in platelet supernatant（

0.05，

0.01）. There was no significant change in the content of TGF-

in SKOV3 supernatant treated with Danshen injection. The platelet aggregation， thromboxane A

（TXB

）， and serotonin （5-HT） secretion in the SKOV3 cell supernatant induction group were significantly increased as compared with those in the blank group （

0.01）， while such indexes in the cell supernatant induction + Danshen injection group were obviously decreased （

0.01）. Compared with the Danshen injection （24 g·L

-1

） group， the cell supernatant induction + Danshen injection group displayed more obvious inhibition at the same dose（

0.01）. Compared with the blank group， the platelet induction group exhibited obviously up-regulated phosphorylated TGF-

-activated kinase-1 （TAK-1） and NF-

B， but down-regulated phosphorylated inhibitory protein of NF-

B （I

B）（

0.01）， which however were significantly reversed in the platelet + Danshen injection group

（

0.01）.

Conclusion

Danshen injection affect the proliferation of SKOV3 cells by inhibiting their interaction with platelets， which may be related to the inhibited secretion of TGF-

关键词

Keywords

references

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Danshen Injection Inhibits SKOV3 Cell Proliferation in Vitro by Interfering with Their Interaction with Platelets

DOI：10.13422/j.cnki.syfjx.20212324

摘要

Abstract

关键词

Keywords

references