Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia

Sen-lin CHEN; Shi-xiang KUANG; Jian-fei LIU; Zhi-lan ZHAO; Xiong ZHENG; Jin ZOU; Shuang LIU

doi:10.13422/j.cnki.syfjx.20220396

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Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia

更新时间：2022-01-13

- Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 4, Pages: 68-75(2022)
- 作者机构：
  
  1.贵州中医药大学，贵阳 550025
  2.贵州中医药大学第二附属医院，贵阳 550003
  3.首都医科大学，北京 100069
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20220396
  CLC： R22;R242;R285.5;R2-031;R746.1;R255.6
- Received：18 April 2021，
  
  Published Online：06 December 2021，
  
  Published：20 February 2022
- 稿件说明：
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陈森林,况时祥,刘建飞等.益气解毒复方含药血清对胸腺增生型重症肌无力mTEC和Treg细胞增殖的影响[J].中国实验方剂学杂志,2022,28(04):68-75.

CHEN Sen-lin,KUANG Shi-xiang,LIU Jian-fei,et al.Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(04):68-75.
陈森林,况时祥,刘建飞等.益气解毒复方含药血清对胸腺增生型重症肌无力mTEC和Treg细胞增殖的影响[J].中国实验方剂学杂志,2022,28(04):68-75. DOI： 10.13422/j.cnki.syfjx.20220396.

CHEN Sen-lin,KUANG Shi-xiang,LIU Jian-fei,et al.Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(04):68-75. DOI： 10.13422/j.cnki.syfjx.20220396.

摘要

目的

探讨益气解毒复方含药血清对胸腺增生型重症肌无力（MG）患者胸腺髓质上皮细胞（mTEC）和调节性T细胞（Treg）增殖的影响。

方法

采用血清药理学方法，将35只SD大鼠适应性喂养1周后随机分为益气解毒复方低、中、高剂量组，空白组，强的松组，每组7只。大鼠连续灌胃1周，制备上述各组含药血清。利用细胞增殖与活性检测（CCK-8）法观察不同浓度益气解毒复方含药血清对mTEC和Treg细胞增殖的作用，并通过共同培养方式观察mTEC及益气解毒复方含药血清对Treg细胞增殖的作用。

结果

胸腺细胞经培育，使用mTEC特征性标记荆豆凝集素Ⅰ（UEAI）做流式检测阳性率均值为92.54%。采用磁珠分选Treg细胞，多次磁珠分选后的Treg细胞纯度高达92%，mTEC和Treg细胞均呈较高阳性率表达，细胞为mTEC和Treg细胞，细胞纯度符合后续实验的要求。益气解毒复方含药血清体积分数2.5%~15%时，对mTEC和Treg细胞均有抑制作用，益气解毒复方含药血清浓度≥20%时对细胞的增殖有促进作用，且随作用的时间延长，含药血清对细胞的增殖的促进作用越为明显。培养48 h后的结果显示，与空白组比较，强的松组和益气解毒复方低剂量组差异无统计学意义，中、高剂量组细胞增殖抑制率显著降低（

0.01）。与mTEC共同培养的Treg细胞，与空白组比较，强的松组、益气解毒复方低剂量组Treg细胞增殖抑制率差异无统计学意义，中、高剂量组细胞增殖抑制率显著降低（

0.01）。

结论

益气解毒复方含药血清能够影响胸腺增生型MG患者mTEC和Treg细胞的增殖，独立培养MG患者的Treg细胞相比，与mTEC共同培养的MG患者Treg细胞增殖具有促进作用，提示mTEC能够调控Treg细胞的增殖，且这种作用在予以益气解毒复方含药血清干预之后更为明显，说明益气解毒复方对Treg细胞的干预，可以通过对mTEC的治疗作用产生，这可能是益气解毒复方治疗MG的作用机制之一。

Abstract

Objective

To investigate the effect of Yiqi Jiedu prescription-containing serum on the proliferation of medullary thymic epithelial cells （mTEC） and regulatory T （Treg） cells in myasthenia gravis （MG） patients with thymus hyperplasia.

Method

According to serological methods，35 SD rats were adaptively fed for one week and randomized into the low-，medium-， and high-dose Yiqi Jiedu prescription groups，control group， and prednisone group，with seven rats in each group， which were then gavaged with the corresponding drugs for one week for preparing the drug-containing serum. The effect of Yiqi Jiedu prescription-containing serum at different concentrations on the proliferation of mTEC and Treg cells were determined by cell counting kit-8 （CCK-8） assay. Besides， the effect of mTEC and Yiqi Jiedu prescription-containing serum on Treg cell proliferation were observed through co-culture.

Result

Thymocytes were cultured for a period of time. Their mean positive rate revealed by flow cytometry using mTEC characteristic marker Ulex europaeus agglutinin Ⅰ （UEAI） was 92.54%. Treg cells were sorted by magnetic beads. The purity of Treg cells after repeated magnetic bead sorting was as high as 92%. mTEC and Treg cells showed high positive expression rates，and their cell purity met the requirements of subsequent experiments. When the concentration of Yiqi Jiedu prescription-containing serum was 2.5%-15%，it exhibited an inhibitory effect against mTEC and Treg cells. When the concentration was equal to or greater than 20%，it promoted cell proliferation，which was further enhanced with the extension of action time. The results after 48 h of culture showed that compared with the control group，prednisone and low-dose Yiqi Jiedu prescription had no significant effect on the proliferation of these two kinds of cells，but the medium- and high-dose Yiqi Jiedu prescription remarkably reduced their proliferation inhibition rate （

0.01）. After co-culture with mTEC， the control group was not significantly different from the prednisone group and the low-dose Yiqi Jiedu prescription-containing serum group in the proliferation of Treg cells，while the medium- and high-dose Yiqi Jiedu prescription-containing serum groups significantly lowered the proliferation inhibition rate （

0.01）.

Conclusion

Yiqi Jiedu prescription-containing serum affects the proliferation of mTEC and Treg cells in MG patients with thymus hyperplasia. Compared with the solely cultured Treg cells isolated from MG patients，the Treg cells co-cultured with mTEC exhibit enhanced proliferation in MG patients，suggesting that mTEC can regulate the proliferation of Treg cells. This effect becomes more obvious after the intervention with Yiqi Jiedu prescription-containing serum，indicating that intervention effect of Yiqi Jiedu prescription on Treg cells can be produced during its treatment of mTEC， which may be one of the mechanisms of Yiqi Jiedu prescription-containing serum in alleviating MG.

关键词

Keywords

references

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