基于TMT蛋白质组学探讨黄精多糖对顺铂诱导C2C12细胞肌管萎缩的作用机制
Effects of
Polygonatum cyrtonema Hua polysaccharide on C2C12 cells with cisplatin-induced myotube atrophy based on TMT proteomics- 北京中医药大学学报 2023年46卷第6期 页码：790-800
- 作者机构：
1.湖南中医药大学科技创新中心 长沙 410208
2.湖南省中医药研究院中药资源研究所
3.湖南省中医药研究院附属医院
4.湖南省中医药研究院医学实验动物中心
- 作者简介：
唐雪阳，女，在读博士生
# 张水寒，女，博士，研究员，博士生导师，主要研究方向：中药资源、制剂及其质量标准，E-mail：zhangshuihan0220@126.com
- 基金信息：国家自然科学基金面上项目(81673585);湖南创新型省份建设专项(2021SK2012)
DOI：10.3969/j.issn.1006-2157.2023.06.009
中图分类号： R285.5纸质出版日期：2023-06-30，
网络出版日期：2023-05-18，
收稿日期：2023-01-08，
- 稿件说明：
移动端阅览
唐雪阳, 周融融, 方晓阳, 等. 基于TMT蛋白质组学探讨黄精多糖对顺铂诱导C2C12细胞肌管萎缩的作用机制[J]. 北京中医药大学学报, 2023,46(6):790-800.
TANG Xueyang, ZHOU Rongrong, FANG Xiaoyang, et al. Effects of
Polygonatum cyrtonema Hua polysaccharide on C2C12 cells with cisplatin-induced myotube atrophy based on TMT proteomics[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(6):790-800.唐雪阳, 周融融, 方晓阳, 等. 基于TMT蛋白质组学探讨黄精多糖对顺铂诱导C2C12细胞肌管萎缩的作用机制[J]. 北京中医药大学学报, 2023,46(6):790-800. DOI： 10.3969/j.issn.1006-2157.2023.06.009.
TANG Xueyang, ZHOU Rongrong, FANG Xiaoyang, et al. Effects of
Polygonatum cyrtonema Hua polysaccharide on C2C12 cells with cisplatin-induced myotube atrophy based on TMT proteomics[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(6):790-800. DOI： 10.3969/j.issn.1006-2157.2023.06.009.
摘要
目的
2
基于串联质量标签(TMT)标记的定量蛋白质组学研究黄精多糖改善顺铂诱导小鼠成肌细胞(C2C12细胞)肌管萎缩的蛋白质代谢轮廓，探讨黄精多糖的作用机制。
方法
2
将C2C12细胞培养至肌管，分为4组：正常组、顺铂组、黄精多糖组、顺铂＋黄精多糖组。按如下方法处理。正常组：不含血清的高糖DMEM培养基；顺铂组：不含血清的高糖DMEM培养基＋50 μmol/L顺铂；黄精多糖组：不含血清的高糖DMEM培养基＋0.200 mg/L黄精多糖；顺铂＋黄精多糖组：不含血清的高糖DMEM培养基＋50 μmol/L顺铂＋0.200 mg/L黄精多糖。各组均避光培养24 h。CCK8法检测C2C12细胞活力，免疫荧光法检测肌管直径和细胞融合指数，TMT蛋白质组学表征黄精多糖改善顺铂致C2C12细胞肌管萎缩的蛋白质代谢轮廓。
结果
2
CCK8法结果显示，黄精多糖可促进C2C12细胞活力，顺铂致C2C12肌管萎缩的造模浓度为50 μmol/L，黄精多糖的给药质量浓度为0.200 mg/L。免疫荧光法结果显示，与顺铂组比较，顺铂＋黄精多糖组萎缩肌管的直径和融合指数升高(
P
<
0.01)。蛋白质组学结果显示，以磷酸泛酰－半胱氨酸连接酶、双功能损伤控制性磷酸酶、2－磷酸－D－甘油酸水解酶、CCR4－NOT转录复合体亚基9为代表的43个蛋白在正常组、顺铂组和顺铂＋黄精多糖组3组间发生了连续性的调节变化，核糖体、鞘脂类代谢、鞘糖脂生物合成相关代谢通路调节在黄精多糖改善顺铂所致的C2C12肌管萎缩中作用明显。
结论
2
黄精多糖能改善顺铂所致C2C12肌管萎缩，其机制可能与下调鞘磷脂磷酸二酯酶2介导的神经酰胺积累有关。
Abstract
Objective
2
To investigate the protein metabolism profile of
Polygonatum cyrtonema
Hua polysaccharide (PCP) on cisplatin-induced myotube atrophy in mouse myoblasts (C2C12 cells) by tandem mass tag (TMT)-labeled quantitative proteomics
and to further explore the mechanism of PCP.
Methods
2
C2C12 cells were cultured into myotubes and divided into four groups: normal group
cisplatin group
PCP group
and cisplatin + PCP group. The cells in the normal group were administered serum-free high-glucose DMEM medium
the cells in the cisplatin group were administered serum-free high-glucose DMEM medium + 50 μmol/L cisplatin
the cells in the PCP group were administered serum-free high-glucose DMEM medium+ 0.200 mg/L PCP
the cells in the cisplatin + PCP group were administered serum-free high-glucose DMEM medium+ 50 μmol/L cisplatin + 0.200 mg/L PCP
and cells of all of the groups were cultured for 24 h in the dark. The viability of C2C12 cells was detected by the CCK8 method
the myotube diameter and cell fusion index were determined by an immunofluorescence method
and the protein metabolism profile of PCP that improved C2C12 myotube atrophy induced by cisplatin was characterized by TMT proteomics.
Results
2
CCK8 result showed that PCP promoted the viability of C2C12 cells
the modeling dose of cisplatin-induced C2C12 myotube atrophy was 50 μmol/L
and the mass concentration of PCP was 0.200 mg/L. The result of immunofluorescence showed that
compared with the cisplatin group
the diameter and fusion index of the myotubes were increased after PCP intervention (
P
<
0.01). Proteomic result showed that 43 proteins
including phosphopantothenate-cysteine ligase
pantothenate kinase 4
α-enolase and CCR4-NOT transcription complex subunit 9
underwent continuous regulatory changes among the normal group
cisplatin group
and cisplatin + PCP group. Regulation of metabolic pathways related to ribosomes
sphingolipid metabolism and sphingolipid biosynthesis contributed to the improvement of C2C12 myotube atrophy induced by cisplatin.
Conclusion
2
PCP improved C2C12 myotube atrophy induced by cisplatin
and its mechanism may be related to downregulating the accumulation of ceramide
which may be mediated by sphingomyelin phosphodiesterase 2.
关键词
黄精多糖成肌细胞顺铂萎缩蛋白质组学
Keywords
Polygonatum cyrtonema Huapolysaccharidemyoblastcisplatinatrophyproteomics
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