芍药苷调控JAK/STAT3通路干预HepG2细胞PD-L1表达的研究

万南燕; 蒋翠花; 高萌; 张健; 殷志琦; 潘珂

doi:10.11665/j.issn.1000-5048.20190213

芍药苷调控JAK/STAT3通路干预HepG2细胞PD-L1表达的研究

万南燕^1,2,3,4,
蒋翠花^2,3,
高萌^2,3,
张健^2,3,
殷志琦⁴,
潘珂¹

1.
中国药科大学中药学院天然药物化学系, 南京 210009
2.
江苏省中医药研究院转化医学实验室, 南京 210028
3.
南京中医药大学附属中西医结合医院南京 210028
4.
中国药科大学中药学院中药制药系&天然药物活性组分与药效国家重点实验室, 南京 210009

基金项目: 江苏高校优势学科建设工程资助项目

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- 刊出日期: 2019-04-24

Paeoniflorin inhibits programmed cell death-1-ligand 1 expression in HepG2 cells by regulating JAK/STAT3 signal pathway

WAN Nanyan^1,2,3,4,
JIANG Cuihua^2,3,
GAO Meng^2,3,
ZHANG Jian^2,3,
YIN Zhiqi⁴,
PAN Ke¹

摘要

摘要: 为探究芍药苷对程序性死亡受体-配体1(programmed cell death-Ligand 1，PD-L1)表达的影响及作用机制，研究采用干扰素γ(IFN-γ)诱导HepG2细胞建立体外PD-L1高表达细胞模型。采用MTT法检测芍药苷的细胞毒性，通过流式细胞术、ELISA、RT-PCR法检测其对PD-L1蛋白和mRNA表达的影响；建立HepG2细胞和Jurkat T细胞共培养体系，ELISA检测芍药苷干预后共培养24 h后的IL-2的表达，CCK-8法检测药物干预后T细胞增殖情况，Western blot检测芍药苷作用后HepG2细胞中PD-L1、Janus激酶(Janus kinase，JAK)和信号传导及转录激活因子(signal transducers and activators of transcription，STAT3)的蛋白表达。实验结果表明，芍药苷能够显著下调PD-L1蛋白和mRNA的表达水平，增加共培养体系中的IL-2的浓度，促进T细胞显著增殖，此外，芍药苷能显著抑制JAK和STAT3的蛋白磷酸化。实验结果表明，芍药苷能够下调PD-L1的表达，其机制可能与JAK/STAT3通路有关。
- 芍药苷 /
- 程序性死亡受体-配体1 /
- 肿瘤免疫逃逸 /
- 免疫检查点
Abstract: In order to explore the effect and its mechanism of paeoniflorin on PD-L1, a PD-L1 high expression cell model was established in interferon gamma(IFN-γ)-induced HepG2 cells. The cytotoxicity of paeoniflorin was detected by MTT assay. Flow cytometry, ELISA and RT-PCR were performed to detect protein and mRNA levels of PD-L1 regulated by paeoniflorin. In HepG2 cells and Jurkat T cell co-culture system, the expression of IL-2 was detected by ELISA. Besides, T cell proliferation was evaluated by CCK-8 method, and the protein expression levels of PD-L1, JAK and STAT3 after drug treatment were determined by Western blot. These results indicated that paeoniflorin could significantly down-regulate the levels of PD-L1 protein and mRNA. In addition, it increased the number of T cells and the concentration of IL-2 in the co-culture system. Furthermore, paeoniflorin could significantly inhibit the protein expression of JAK and STAT3. Au the above experimental data indicated that paeoniflorin could down-regulate the expression of PD-L1, and its mechanism might be related to the JAK/STAT3 pathway.
- paeoniflorin /
- PD-L1 /
- tumor immune escape /
- immune checkpoint

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芍药苷调控JAK/STAT3通路干预HepG2细胞PD-L1表达的研究

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Paeoniflorin inhibits programmed cell death-1-ligand 1 expression in HepG2 cells by regulating JAK/STAT3 signal pathway

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