清毒栓对宫颈癌细胞凋亡及Th1/Th2、Th17/Treg平衡调控的影响

耿韦华; 楼姣英; 王莹; 余碧波

doi:10.3969/j.issn.2095-6606.2023.04.022

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清毒栓对宫颈癌细胞凋亡及Th1/Th2、Th17/Treg平衡调控的影响

中医临床基础研究 | 更新时间：2023-08-07

- 清毒栓对宫颈癌细胞凋亡及Th1/Th2、Th17/Treg平衡调控的影响
- Effect of Qingdu Suppository on apoptosis of cervical cancer cells and regulation of Th1/Th2 and Th17/Treg
- 现代中医临床 2023年30卷第4期页码：99-106
- 作者机构：
  
  1.北京中医药大学东方医院　北京　100078
  2.北京中西医结合医院
  3.北京市大兴区妇幼保健院
- 作者简介：
  
  耿韦华，女，在读博士生
  # 楼姣英，女，博士，主任医师、教授，博士生导师，E-mail：doctorlou@126.com
- 基金信息：
  
  国家自然科学基金面上项目(81473515)
- DOI：10.3969/j.issn.2095-6606.2023.04.022
  中图分类号： R285.5
- 纸质出版日期：2023-07-30，
  
  收稿日期：2022-04-12，
- 稿件说明：
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耿韦华, 楼姣英, 王莹, 等. 清毒栓对宫颈癌细胞凋亡及Th1/Th2、Th17/Treg平衡调控的影响[J]. 现代中医临床, 2023,30(4):99-106.

GENG Weihua, LOU Jiaoying, WANG Ying, et al. Effect of Qingdu Suppository on apoptosis of cervical cancer cells and regulation of Th1/Th2 and Th17/Treg[J]. Modern Chinese Clinical Medicine, 2023,30(4):99-106.
耿韦华, 楼姣英, 王莹, 等. 清毒栓对宫颈癌细胞凋亡及Th1/Th2、Th17/Treg平衡调控的影响[J]. 现代中医临床, 2023,30(4):99-106. DOI： 10.3969/j.issn.2095-6606.2023.04.022.

GENG Weihua, LOU Jiaoying, WANG Ying, et al. Effect of Qingdu Suppository on apoptosis of cervical cancer cells and regulation of Th1/Th2 and Th17/Treg[J]. Modern Chinese Clinical Medicine, 2023,30(4):99-106. DOI： 10.3969/j.issn.2095-6606.2023.04.022.

摘要

目的

基于细胞凋亡及免疫调控探讨清毒栓体内外抗宫颈癌的作用及机制。

方法

(1)细胞实验：将宫颈癌SiHa细胞分为5%、10%、15%、20%空白血清组；5%、10%、15%、20%含药血清组，CCK8法测定不同浓度血清对SiHa细胞活性的影响。流式细胞技术检测15%含药血清处理SiHa细胞72 h后，细胞凋亡及周期分布情况。(2)动物实验：构建宫颈癌荷瘤小鼠模型，随机数字表法将小鼠分为宫颈癌对照组(磷酸盐缓冲液灌胃)、清毒栓低剂量组(14.4 g/kg中药灌胃)、清毒栓高剂量组(28.8 g/kg中药灌胃)，每组6只，连续给药28 d，记录并计算瘤体积和体质量。分别用Ki67法和Tunel法检测3组小鼠癌组织增殖、凋亡，并用流式检测3组小鼠外周血单个核细胞中T细胞亚型表达。

结果

细胞实验表明含药血清可以抑制SiHa细胞活性，其中15%和20%含药血清处理72 h时，抑制作用最强烈，且差异无统计学意义。与15%空白血清组相比，15%含药血清组凋亡细胞增多；15%含药血清增加G0/G1期细胞，减少S期和G2/M期细胞，差异均具有统计学意义(

0.01)。动物实验表明相较于宫颈癌对照组，清毒栓组癌组织中Ki67阳性细胞比例降低、Tunel阳性细胞比例增加，且高剂量组降低或增加更明显，差异均有统计学意义(

0.01)。相较于宫颈癌对照组，清毒栓高剂量组小鼠外周血中Th1、Th17细胞比例增加，Th2、Treg细胞比例降低，Th1/Th2、Th17/Treg比例增加，差异均有统计学意义(

0.05)。

结论

清毒栓抑制SiHa细胞活性的最佳剂量是15%的含药血清，作用72 h。清毒栓可能是通过促进癌细胞凋亡，或通过免疫微环境，调控Th2向Th1漂移，抑制Treg细胞，解除免疫抑制，从而实现抑制宫颈癌。

Abstract

Objective

To investigate the effect and mechanism of Qingdu Suppository (Toxin-Clearing Suppository) on cervical cancer

in vitro

and

in vivo

based on cell apoptosis and immune regulation.

Methods

(1) Cell experiment: SiHa cells were divided into blank serum groups at various concentrations (5%

10%

15%

20%

respectively) and dosed serum groups at various concentrations (5%

10%

15%

20%

respectively). The CCK8 method was employed to investigate the effects of different serum concentrations on SiHa cell activity. Flow cytometry was then used to detect the apoptosis and cycle distribution of the cells treated with 15% dosed serum for 72 h. (2) Animal experiment: A tumor bearing mouse model of cervical cancer was established. The mice were divided into cervical cancer control group (PBS buffer gavage)

Qingdu Suppository low-dosage group (14.4 g/kg gavage) and Qingdu Suppository high-dosage group (28.8 g/kg gavage) by random number table method

with 6 mice in each group. The intervention lasted for 28 days. The tumor volume and body weight were recorded. The proliferation and apoptosis of tumor tissues in the three groups were detected via Ki67 assay and Tunel assay

respectively

and the expression of T cell subtypes in tumor tissues of the three groups were detected by flow cytometry.

Results

The cell experiment showed that the dosed serum could inhibit the activity of SiHa cells

with the 15% and 20% dosed serum exhibiting the strongest inhibitory effect when the cells were treated for 72 h

and there was no significant difference between the two groups. Compared with the 15% blank serum group

apoptotic cells increased in the 15% dosed serum group. Cells in phase G0/G1 increased and those in phase S and G2/M decreased in dosed serum

the differences were statistically significant (

0.01). The animal experiment showed that compared with the control group

the proportion of Ki67 positive cells in the tumor tissues of the two Qingdu Suppository groups decreased while Tunel positive cells increased

and the changes in the high-dosage group were more prominent with the differences being statistically significant between the two groups (

0.01). Compared with the control group

the proportion of Th1 and Th17 cells in peripheral blood of mice in the high-dosage group increased

the proportion of Th2 and Treg cells decreased

and the proportion of Th1/Th2 and Th17/Treg increased

with the differences being statistically significant (

0.05).

Conclusion

The optimal dosage of Qingdu Suppository to inhibit SiHa cell activity is 15% for 72 hours. Qingdu Suppository may inhibit cervical cancer by promoting apoptosis of cancer cells. This may also be achieved by regulating drift of Th2 to Th1

inhibiting Treg cells and relieving immunosuppression through immune microenvironment.

关键词

清毒栓宫颈癌凋亡Th1/Th2Th17/Treg

Keywords

Qingdu Suppositorycervical cancerapoptosisTh1/Th2Th17/Treg

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