轴突导向因子受体3缺陷促进化疗药物诱导的巨噬细胞泡沫化进程

doi:10.3969/j.issn.1006-5725.2024.06.010

摘要/Abstract

摘要：

目的探索化疗药物阿霉素或顺铂对巨噬细胞脂质代谢的影响及调控机制。方法阿霉素或者顺铂处理巨噬细胞RAW264.7，用油红O、ELISA等检测细胞内脂质水平；用RNA sequence和Western blot筛选和验证化疗药物处理后的基因表达变化；探讨沉默ROBO3对细胞脂质代谢的影响，并用Q-PCR和Western blot检测脂质代谢关键靶基因的变化。结果阿霉素或顺铂可诱发巨噬细胞胆固醇代谢紊乱，加剧巨噬细胞泡沫化。进一步研究显示，轴突导向因子受体ROBO3的表达水平在化疗药物诱导巨噬细胞泡沫化进程中先增高后降低；沉默ROBO3加重oxldl诱导的巨噬细胞泡沫化水平。机制上，ROBO3沉默可上升胆固醇合成相关基因DHCR24表达，抑制胆固醇外排相关基因ABCG1表达，造成巨噬细胞内胆固醇蓄积。结论 ROBO3在化疗药物诱导巨噬细胞胆固醇代谢紊乱及泡沫化进程中发挥重要调控作用，可为化疗相关动脉粥样硬化的防治提供新的靶点和思路。

关键词: 化疗血管毒性, 巨噬-泡沫细胞, 轴突导向因子受体3, 胆固醇代谢

Abstract:

Objective To explore the effect of chemotherapeutic drugs doxorubicin or cisplatin on lipid metabolism of macrophages and its regulatory mechanism. Methods Macrophage RAW264.7 was treated with doxorubicin or cisplatin， and intracellular lipid level was detected by oil red O and ELISA； RNA sequence screening and Western blot were used to confirm the changes of gene expression after chemotherapeutic drug treatment； The effects of silencing ROBO3 on cellular lipid metabolism were explored， and changes in key target genes of lipid metabolism were detected by Q-PCR and western blot. Results Adriamycin or cisplatin induced disturbances in macrophage cholesterol metabolism and exacerbated macrophage foaminess. Further studies showed that the expression of the axon guidance factor receptor， ROBO3， increased and then decreased during the chemotherapeutic drug-induced macrophage foaming process. Further intervention with ROBO3 exacerbates oxldl-induced cholesterol accumulation and foam formation in macrophages. Mechanistically， ROBO3 deficiency promotes the expression of cholesterol synthesis-related gene DHCR24 and inhibits the expression of cholesterol elimination-related gene ABCG1， resulting in cholesterol accumulation in macrophages. Conclusion This study found that ROBO3 plays an important regulatory role in the disruption of cholesterol metabolism and its foaming process in macrophages induced by chemotherapeutic drugs， which may provide new targets and ideas for the prevention and treatment of chemotherapy-related atherosclerosis.

Key words: chemotherapy vascular toxicity, macrophages-foam cells, ROBO3, cholesterol metabolism

中图分类号:

刘永,程晓雷,崔香丽,唐颢,陈还珍. 轴突导向因子受体3缺陷促进化疗药物诱导的巨噬细胞泡沫化进程[J]. 实用医学杂志, 2024, 40(6): 787-795.

Yong LIU,XiaoLei CHENG,Xiangli CUI,Hao TANG,Huanzhen CHEN. ROBO3 deficiency promotes chemotherapy⁃induced transition of macrophage to foam cell[J]. The Journal of Practical Medicine, 2024, 40(6): 787-795.

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基因名称	上游引物（5′-3′）	下游引物（5′-3′）
ABCA1	GCTTGTTGGCCTCAGTTAAGG	GTAGCTCAGGCGTACAGAGAT
ABCG1	GGCAGACGAGAGATGGTCAAA	AACATGACAGGCGGGTTGTT
DHCR24	GAGCCCAGCAAGCTGAATAG	GGTTGTTGCCGAAAGGGATG
NCEH1	CGCAGCAAGTGAGTAACCTGATA	GCTGATCTTTGCACTAGCCAGG
NPC1	ACCAAAGGAAGCCGAGTGAG	TATTCACCGACGGCCCTATG
SOAT1	TCGCAGGAAGCTTGATTGATAG	AAATAGTGGCTTCAGCTCCTCC
SOAT2	TTGGTGCAATGGACTCGACA	GGTGTAACTCTTGGGTCTTGC
SRA	CCAGCAATGACAAAAGAGATGACA	CTGAAGGGAGGGGCCATTTT
SRB1	GGTGCTCAAGAATGTCCGCAT	GTAGAAAGGGACGGGGATCTC
ACTIN	GTGACGTTGACATCCGTAAAGA	GCCGGACTCATCGTACTCC

基因名	log₂fc（Cpt/NC）	log₂fc（DOX/NC）	NC vs. DOX regulate
Robo3	9.32522464	5.850870275	up
Exosc6	8.767319376	9.572233415	up
Ces2g	7.981918639	5.31450618	up
Dclk2	7.294684025	6.128071326	up
Sec1415	7.09685857	5.121969243	up
H2-Eb1	6.936395687	5.618841914	up
Gm15987	6.577097143	6.32926248	up
Col6a3	6.296195482	8.103776317	up
Urah	5.755490065	5.407129491	up
Sgca	5.43349494	7.200322844	up
Ccl2	5.375203607	5.38460591	up
Camsap3	5.032565	5.567130258	up