浏览全部资源
扫码关注微信
南京中医药大学药学院,南京 210023
硕士研究生。研究方向:免疫药理学。E-mail:2380189936@qq.com
教授,博士生导师,博士。研究方向:免疫药理学、中药药理学。电话:025-5811933。E-mail:llzhou74@163.com
纸质出版日期:2024-01-15,
收稿日期:2023-07-15,
修回日期:2023-12-05,
扫 描 看 全 文
葛玉,陈雪,王福荣等.梓醇干预PKM2/LDHA表达调控Th17细胞分化的机制研究 Δ[J].中国药房,2024,35(01):15-20.
GE Yu,CHEN Xue,WANG Furong,et al.Mechanism of catalpol regulating Th17 cell differentiation by interfering PKM2/LDHA expression[J].ZHONGGUO YAOFANG,2024,35(01):15-20.
葛玉,陈雪,王福荣等.梓醇干预PKM2/LDHA表达调控Th17细胞分化的机制研究 Δ[J].中国药房,2024,35(01):15-20. DOI: 10.6039/j.issn.1001-0408.2024.01.03.
GE Yu,CHEN Xue,WANG Furong,et al.Mechanism of catalpol regulating Th17 cell differentiation by interfering PKM2/LDHA expression[J].ZHONGGUO YAOFANG,2024,35(01):15-20. DOI: 10.6039/j.issn.1001-0408.2024.01.03.
目的
2
研究梓醇通过干预丙酮酸激酶M2(PKM2)、乳酸脱氢酶A(LDHA)表达进而影响辅助性T细胞17(Th17)分化的机制。
方法
2
从C57BL/6小鼠脾脏中分选出naive CD4
+
T细胞,通过加入定向分化刺激剂诱导72 h使naive CD4
+
T细胞向Th17细胞分化。在诱导分化的同时,分别用0(定向对照)、20、40、80 μg/mL梓醇对细胞进行处理。采用流式细胞术检测细胞中Th17细胞分化比例,采用比色法检测细胞培养上清液中丙酮酸、乳酸水平,采用实时荧光定量-逆转录聚合酶链式反应(qRT-PCR)法检测细胞中视黄酸相关孤儿受体γt(RORγt)、PKM2、LDHA mRNA表达情况,采用Western blot法检测细胞中PKM2、LDHA、信号转导和转录激活因子3(STAT3)、磷酸化STAT3(p-STAT3)蛋白表达情况。
结果
2
与定向对照组比较,经20、40、80 μg/mL梓醇作用72 h后,细胞中Th17细胞分化比例分别降低了6.74%、8.41%、9.24%;细胞培养上清液中丙酮酸、乳酸水平,细胞中PKM2、LDHA、RORγt
mRNA表达水平以及细胞中PKM2、LDHA蛋白表达水平和STAT3磷酸化水平均显著降低(
P
<0.05)。
结论
2
梓醇可通过下调PKM2、LDHA表达来降低糖酵解水平,进而抑制Th17细胞分化。
OBJECTIVE
2
To investigate the mechanism of catalpol affecting the differentiation of helper T cell 17 (Th17) by interfering the expressions of pyruvate kinase M2 (PKM2) and lactate dehydrogenase A (LDHA).
METHODS
2
The naive CD4
+
T cells were selected from the spleen of C57BL/6 mice, and were differentiated into Th17 cells by adding directional differentiation stimulants for 72 hours. At the same time, the cells were treated with 0 (directed control), 20, 40 and 80 μg/mL catalpol. The flow cytometry was used to detect the proportion of Th17 cell differentiation in cells; the colorimetric method was adopted to detect the levels of pyruvate and lactate in cell culture supernatant; mRNA expressions of retinoid-related orphan nuclear receptor gamma t (RORγt), PKM2 and LDHA were detected by qRT-PCR method; Western blot was used to detect the expression levels of PKM2, LDHA, signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 (p-STAT3) proteins in cells.
RESULTS
2
Compared with the directed control group, after 72 hours of treatment with 20, 40, 80 μg/mL catalpol, the differentiation ratio of Th17 cells were decreased by 6.74%, 8.41%, 9.24%, and the levels of pyruvate and lactate in the cell culture supernatant, the mRNA expressions of PKM2, LDHA and RORγt as well as the protein expressions of PKM2 and LDHA and the phosphorylation of STAT3 were significantly reduced (
P
<0.05).
CONCLUSIONS
2
Catalpol can reduce the glycolysis level by down-regulating the expressions of PKM2 and LDHA, thereby inhibiting the differentiation of Th17 cells.
梓醇糖酵解辅助性T细胞17丙酮酸激酶M2乳酸脱氢酶A
glycolysishelper T cell 17pyruvate kinase M2lactate dehydrogenase A
童荣生. 生物制剂治疗类风湿关节炎合理用药中国专家共识[J]. 中国新药杂志,2022,31(21):2174-2184.
TONG R S. Chinese expert consensus on rational use of biological agents in the treatment of rheumatoid arthritis[J]. Chin J N Drugs,2022,31(21):2174-2184.
PARK H,LI Z X,YANG X O,et al. A distinct lineage of CD4+ T cells regulates tissue inflammation by producing interleukin 17[J]. Nat Immunol,2005,6(11):1133-1141.
YANG P,QIAN F Y,ZHANG M F,et al. Th17 cell pathogenicity and plasticity in rheumatoid arthritis[J]. J Leukoc Biol,2019,106(6):1233-1240.
MILOVANOVIC J,ARSENIJEVIC A,STOJANOVIC B,et al. Interleukin-17 in chronic inflammatory neurological diseases[J]. Frot Immunol,2020,11:947.
庞文翼. PKM2调节糖酵解和细胞外基质沉积促进CTEPH血管重构的实验研究[D]. 北京:北京协和医学院,2021.
PANG W Y. PKM2 regulates glycolysis and extracellular matrix to promote CTEPH vascular remodeling[D]. Beijing:Peking Union Medical College,2021.
CAI W W,YU Y,ZONG S Y,et al. Metabolic reprogramming as a key regulator in the pathogenesis of rheumatoid arthritis[J]. Inflamm Res,2020,69(11):1087-1101.
PUCINO V,BOMBARDIERI M,PITZALIS C,et al. Lactate at the crossroads of metabolism,inflammation,and autoimmunity[J]. Eur J Immunol,2017,47(1):14-21.
朱珏,朱香梅,石雨荷,等. 地黄的研究进展及其质量标志物的预测分析[J]. 中药材,2022,45(5):1273-1281.
ZHU J,ZHU X M,SHI Y H,et al. Research progress of Rehmannia glutinosa and prediction and analysis of its quality markers[J]. J Chin Med Mater,2022,45(5):1273-1281.
钱飞亚,杨培,张明菲,等. 滋肾通络方对胶原性关节炎小鼠Th17/Treg细胞分化及平衡的影响[J]. 中国药理学通报,2019,35(5):720-726.
QIAN F Y,YANG P,ZHANG M F,et al. Effect of ZishenTongluo formula on differentiation and balance of Th17/Treg cells in collagen-induced arthritis[J]. Chin Pharmacol Bull,2019,35(5):720-726.
申美玉,荣秋妮,狄昱希,等. 梓醇调控miR-143-3p干预Th17糖酵解和细胞分化的机制[J]. 中国药理学通报,2022,38(7):1072-1081.
SHEN M Y,RONG Q N,DI Y X,et al. Mechanisms of catalpol inhibiting differentiation and glycolysis of Th17 cells through miR-143-3p[J]. Chin Pharmacol Bull,2022,38(7):1072-1081.
WANG Z H,ZHAN-SHENG H. Catalpol inhibits migration and induces apoptosis in gastric cancer cells and in athymic nude mice[J]. Biomed Pharmacother,2018,103:1708-1719.
PENG H Y,WANG L Q,DAS J K,et al. Control of CD4+ T cells to restrain inflammatory diseases via eukaryotic elongation factor 2 kinase[J]. Signal Transduct Target Ther,2023,8(1):415.
URBAŃSKA K,ORZECHOWSKI A. Unappreciated role of LDHA and LDHB to control apoptosis and autophagy in tumor cells[J]. Int J Mol Sci,2019,20(9):2085.
雷明. 烟酰胺磷酸核糖转移酶和磷酸丙糖异构酶1在类风湿关节炎中的作用及机制研究[D]. 合肥:安徽医科大学,2022.
LEI M. Research on the role and mechanism of nicoti- namide phosphoribosyltransferase and triosephosphate isomerase 1 in rheumatoid arthritis[D]. Hefei:Anhui Medical University,2022.
SHEN M Y,WANG X,DI Y X,et al. Triptolide inhibits Th17 differentiation via controlling PKM2-mediated glycolysis in rheumatoid arthritis[J]. Immunopharmacol Immunotoxicol,2022,44(6):838-849.
WANG G,SU Z H,LI H,et al. The role of metabolism in Th17 cell differentiation and autoimmune diseases[J].Int Immunopharmacol,2022,103:108450.
SHEN H X,SHI L Z.Metabolic regulation of T(H)17 cells[J].Mol Immunol,2019,109:81-87.
KUMAR R,THEISS A L,VENUPRASAD K. RORγt protein modifications and IL-17-mediated inflammation[J].Trends Immunol,2021,42(11):1037-1050.
KIM H,LEE H J,SIM D Y,et al.The antitumor effect of cinnamaldehyde derivative CB-PIC in hepatocellular carcinoma cells via inhibition of pyruvate and STAT3 signaling[J].Int J Mol Sci,2022,23(12):6461.
0
浏览量
12
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构