趋化因子CCL3、CCL4在儿童免疫性血小板减少症中的表达及临床意义

doi:10.12372/jcp.2022.21e1233

摘要/Abstract

摘要：

目的探讨趋化因子CCL3、CCL4在儿童免疫性血小板减少症（ITP）中的表达及其临床意义。方法选择2019年12月至2020年4月住院确诊的ITP患儿为研究对象,根据病程分为新诊断组（n=37,病程≤3个月）和非新诊断组（n=27,病程>3个月）,根据采集标本时机（初诊及入院治疗7天时）分为治疗前组（n=29）和治疗后组（n=35）;并以门诊健康体检儿童为对照组,比较不同组间CCL3、CCL4表达,血小板（PCT）相关指标等。结果共纳入64例ITP患儿,男42例、女22例,中位年龄5.1（3.0~8.0）岁;健康对照组19例,男12例、女7例,中位年龄为4.2（2.8~6.0）岁。治疗前组、治疗后组与对照组之间PLT计数、PLT体积（MPV）、PLT比率（P-LCR）、PLT分布宽度（PDW）、PLT比积（PCT）以及IgG水平的差异均有统计学意义（P<0.05）;治疗前组的PLT计数和PCT较低,MPV、P-LCR、PDW以及IgG较高。新诊断组、非新诊断组CCL3、CCL4浓度均高于对照组,差异有统计学意义（P<0.05）。治疗前组、治疗后组CCL3和CCL4浓度均高于对照组,治疗后组CCL3和 CCL4浓度均低于治疗前组,差异有统计学意义（P<0.05）。ITP不同出血评分组之间CCL3、CCL4表达差异有统计学意义（P<0.05）;随着出血评分增加,CCL3、CCL4表达水平逐渐增加。治疗无效组的CCL3、CCL4表达水平高于完全反应组和反应组,而反应组的CCL3、CCL4表达均低于完全反应组,差异有统计学意义（P<0.05）。结论 CCL3、CCL4可作为评估儿童ITP病情危重程度及治疗反应的分子标志物。

关键词: 免疫性血小板减少症, 趋化因子CCL3, 趋化因子CCL4, 儿童

Abstract:

Objective To investigate expression and clinical significance of chemokines CCL3 and CCL4 in childhood immune thrombocytopenia (ITP). Methods Children with ITP diagnosed in Affiliated Hospital of Zunyi Medical University from December 2019 to April 2020 were selected as study subjects, and were divided into newly diagnosed group (n=37, disease duration ≤3 months) and non-newly diagnosed group (n=27, disease duration >3 months) according to the disease duration, and into pre-treatment group (n=29) and post-treatment group (n=35) according to the timing of specimen collection (at the first diagnosis and 7th day of hospital admission); and healthy children with outpatient health examination were used as the control group, and the expression of CCL3 and CCL4 and platelet-related indexes were compared between different groups. Results A total of 64 children with ITP, 42 males and 22 females, with a median age of 5.1 (3.0-8.0) years, and 19 healthy controls, 12 males and 7 females, with a median age of 4.2 (2.8-6.0) years, were included. The differences in PLT counts, MPV, P-LCR, PDW, PCT, and IgG levels between the pre-treatment group, post-treatment group, and control group were statistically significant (P<0.05); PLT counts and PCT were lower and MPV, P-LCR, PDW, and IgG were higher in the pre-treatment group. CCL3 and CCL4 concentrations were higher in the newly diagnosed and non-newly diagnosed groups than in the control group, with statistically significant differences (P<0.05). The CCL3 and CCL4 concentrations were higher in the pre-treatment and post-treatment groups than in the control group, and the CCL3 and CCL4 concentrations were lower in the post-treatment group than in the pre-treatment group, with statistically significant differences (P<0.05). The differences in CCL3 and CCL4 expression between different bleeding score groups of ITP were statistically significant (P<0.05). The expression levels of CCL3 and CCL4 increased gradually with the increase of bleeding score. The CCL3 and CCL4 expression levels in the treatment ineffective group were higher than those in the complete response group and the response group, while the CCL3 and CCL4 expressions in the response group were lower than those in the complete response group, and the differences were statistically significant (P<0.05). Conclusion CCL3 and CCL4 can be used as children's molecular markers of ITP to assess its severity and response to treatment.

Key words: immune thrombocytopenia, chemokine CCL3, chemokine CCL4, child

陈东平, 罗茜, 黄佩, 郭义敏, 吴柳松, 何志旭, 陈艳. 趋化因子CCL3、CCL4在儿童免疫性血小板减少症中的表达及临床意义[J]. 临床儿科杂志, 2022, 40(2): 95-100.

CHEN Dongping, LUO Xi, HUANG Pei, GUO Yimin, WU Liusong, HE Zhixu, CHEN Yan. Expression and clinical essential of chemokines CCL3 and CCL4 in children with ITP[J]. Journal of Clinical Pediatrics, 2022, 40(2): 95-100.

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参考文献 24

[1]	Harcken C, Kuzmich D, Cook B, et al. Identification of novel azaindazole CCR1 antagonist clinical candidates[J]. Bioorg Med Chem Lett, 2019, 29(3):441-448. doi: 10.1016/j.bmcl.2018.12.024
[2]	Despotovic JM, Grimes AB. Pediatric ITP: is it different from adult ITP?[J]. Hematology Am Soc Hematol Educ Program, 2018, 2018(1):405-411. doi: 10.1182/asheducation-2018.1.405 pmid: 30504339
[3]	Larsen O, Lückmann M, van der Velden W, et al. Selective allosteric modulation of N-terminally cleaved, but not full length CCL3 in CCR1[J]. ACS Pharmacol Transl Sci, 2019, 2(6):429-441. doi: 10.1021/acsptsci.9b00059 pmid: 32259075
[4]	De Meester B, de Vries L, Özparpucu M, et al. Vessel-specific reintroduction of cinnamoyl-CoA reductase1 (CCR1) in dwarfed CCR1 mutants restores vessel and xylary fiber integrity and increases biomass[J]. Plant Physiol, 2018, 176(1):611-633. doi: 10.1104/pp.17.01462
[5]	Sanchez JE, Huma Z, Lane JR, et al. Evaluation and extension of the two-site, two-step model for binding and activation of the chemokine receptor CCR1[J]. J Biol Chem, 2019, 294(10):3464-3475. doi: 10.1074/jbc.RA118.006535 pmid: 30567735
[6]	Yu J, Hua M, Zhao X, et al. NF-κB-94ins/del ATTG genotype contributes to the susceptibility and imbalanced Th17 cells in patients with immune thrombocytopenia[J]. J Immunol Res, 2018: 8170436.
[7]	李宝华, 陈海燕, 石梦娇, 等. 髓源性抑制细胞(MDSC)相关趋化因子MIP-1γ及其受体CCR1在肝癌荷瘤小鼠脾脏的表达及意义[J]. 细胞与分子免疫学杂志, 2020, 36(9):802-808.
[8]	Kim JH, Kim WS, Hong JY, et al. Epstein-Barr virus EBNA2 directs doxorubicin resistance of B cell lymphoma through CCL3 and CCL4-mediated activation of NF-κB and Btk[J]. Oncotarget, 2017, 8(3):5361-5370. doi: 10.18632/oncotarget.v8i3
[9]	王天有, 马洁, 方拥军, 等. 儿童原发性免疫性血小板减少症诊疗规范(2019年版)[J]. 全科医学临床与教育, 2019, 17(12):1059-1062.
[10]	Consolini R, Legitimo A, Caparello MC. The centenary of immune thrombocytopenia - part 1: revising nomenclature and pathogenesis[J]. Front Pediatr, 2016, 4:102. pmid: 27807534
[11]	李蓉蔚, 付荣凤, 陈云飞, 等. 原发免疫性血小板减少症住院患儿临床分析[J]. 中国实验血液学杂志, 2021, 29(2):574-580.
[12]	吕明恩, 李洋, 吕翠翠, 等. 原发免疫性血小板减少症患者出血风险与血小板活化的相关性分析[J]. 中华血液学杂志, 2017, 38(1):33-38.
[13]	Boulware R, Refaai MA. Why do patients with immune thrombocytopenia (ITP) experience lower bleeding events despite thrombocytopenia?[J]. Thromb Res, 2020, 187:154-158. doi: 10.1016/j.thromres.2020.01.020 pmid: 32004875
[14]	李蓉蔚, 付荣凤, 陈云飞, 等. 原发免疫性血小板减少症住院患儿临床分析[J]. 中国实验血液学杂志, 2021. 29(2):574-580.
[15]	Psaila B, Bussel JB, Frelinger AL, et al. Differences in platelet function in patients with acute myeloid leukemia and myelodysplasia compared to equally thrombocytopenic patients with immune thrombocytopenia[J]. J Thromb Haemost, 2011, 9(11):2302-2310. doi: 10.1111/j.1538-7836.2011.04506.x pmid: 21920014
[16]	余强. 血小板参数指标对儿童血小板减少症病因分析的价值[J]. 实用检验医师杂志, 2017, 9(4):217-219.
[17]	黄菊, 卫仿仙, 韩艳霞, 等. 原发性免疫性血小板减少症患者骨髓TPO及c-mpl水平测定意义[J]. 实用医学杂志, 2021, 37(15):1982-1987.
[18]	倪蔚文, 储金华, 杨林海, 等. 儿童初诊原发免疫性血小板减少症患者细胞及体液免疫功能分析[J]. 安徽医科大学学报, 2019, 54(10):1628-1631.
[19]	李昕雨, 何杨, 阮长耿. 原发免疫性血小板减少症的治疗进展[J]. 中国实验血液学杂志, 2021, 29(3):983-987.
[20]	Song Q, Shang J, Zhang C, et al. Transcription factor RUNX3 promotes CD8(+) T cell recruitment by CCL3 and CCL20 in lung adenocarcinoma immune microenvironment[J]. J Cell Biochem, 2020, 121(5-6):3208-3220. doi: 10.1002/jcb.v121.5-6
[21]	Soleimani M, Soleymani A, Seyyedirad N. Elevated CSF concentration of CCL3 and CCL4 in relapsing remitting multiple sclerosis patients[J]. J Immunoassay Immunochem, 2019, 40(4):378-385. doi: 10.1080/15321819.2019.1613242
[22]	De la Fuente López, Landskron G, Parada D, et al. The relationship between chemokines CCL2, CCL3, and CCL4 with the tumor microenvironment and tumor-associated macrophage markers in colorectal cancer[J]. Tumour Biol, 2018, 40(11):1010428318810059.
[23]	Allen F, Bobanga ID, Rauhe P, et al. CCL3 augments tumor rejection and enhances CD8+ T cell infiltration through NK and CD103+ dendritic cell recruitment via IFNγ[J]. Oncoimmunology, 2018, 7(3):e1393598. doi: 10.1080/2162402X.2017.1393598
[24]	吴润晖, 刘雅莉, 高举, 等. 中国儿童原发性免疫性血小板减少症诊断与治疗改编指南(2021版)解读[J]. 中华儿科杂志, 2021, 59(10):820-823.

项目	治疗前组(n=29)	治疗后组(n=35)	对照组(n=19)	统计量	P
PLT[M(P₂₅~P₇₅)]×10⁹·L^–1	9.0(7.0~13.0)¹⁾	62.0(37.0~78.0)¹⁾	236.0(184.5~299.0)	H=59.63	<0.001
WBC分布[n(%)]				χ²=0.04	0.981
（4~10）×10⁹·L^–1	25(86.2)	30(85.7)	16(84.2)
>10×10⁹·L^–1	4(13.8)	5(14.3)	3(15.8)
MPV(x±s)/fl	15.2±0.7¹⁾	13.1±2.4	12.3±2.0	F=15.96	<0.001
P-LCR[M(P₂₅~P₇₅)]/%	45.0(39.5~55.3)¹⁾	33.9(27.4~40.2)	35.7(18.5~39.4)	H=25.53	<0.001
PDW(x±s)/%	19.9±2.1¹⁾	16.1±2.2	15.7±1.7	F=32.76	<0.001
PCT(x±s)/%	0.2±0.1¹⁾	0.4±0.1	0.4±0.1	F=48.92	<0.001
ALT[M(P₂₅~P₇₅)]/U·L^–1	27.7(20.2~32.2)	25.8(17.3~31.4)	23.1(19.9~28.9)	H=1.07	0.585
AST[M(P₂₅~P₇₅)]/U·L^–1	25.1(20.8~35.0)	26.3(18.9~31.3)	26.7(21.2~31.0)	H=0.05	0.974
BUN[M(P₂₅~P₇₅)]/μmol·L^–1	4.7(3.5~5.7)	4.2(3.7~5.3)	4.4(5.4~6.1)	H=4.24	0.120
Cr[M(P₂₅~P₇₅)]/μmol·L^–1	70.2(57.4~83.1)	64.1(55.0~78.5)	64.3(47.2~80.5)	H=2.50	0.286
TG[M(P₂₅~P₇₅)]/mmol·L^–1	1.0(0.8~1.4)	1.1(0.6~1.4)	1.1(0.7~1.4)	H=0.69	0.707
TC[M(P₂₅~P₇₅)]/mmol·L^–1	4.0(3.5~4.8)	3.9(3.3~4.9)	4.2(3.7~4.4)	H=0.77	0.682
LDL[M(P₂₅~P₇₅)/mmol·L^–1	2.5(2.3~2.8)	2.5(2.3~2.7)	2.8(2.3~2.9)	H=2.18	0.336
PT (x±s)/s	12.6±1.0	11.6±1.3	12.3±1.4	F=0.43	0.563
APTT(x±s)/s	31.2±5.0	32.7±4.2	30.2±5.5	F=1.89	0.158
Fib(x±s)/g·L^–1	2.9± 0.7	3.2±0.2	3.0±0.6	F=3.00	0.056
TT(x±s)/s	18.0±1.6	17.4±2.3	18.7±1.8	F=2.98	0.056
IgG(x±s)/g·L^–1	16.5±2.21)	11.7±2.2	11.4±1.9	F=48.48	<0.001
IgA[M(P₂₅~P₇₅)]/g·L^–1	1.9(1.0~2.8)	2.3(1.2~2.7)	2.2(1.6~2.7)	H=0.92	0.632
IgM[M(P₂₅~P₇₅)]/g·L^–1	1.7(1.3~2.1)	1.8(1.3~2.3)	1.8(1.5~2.1)	H=0.75	0.688
IgE[M(P₂₅~P₇₅)]/IU·mL^–1	12.1(10.3~13.8)	12.0(8.2~15.0)	9.9(5.9~13.8)	H=1.95	0.378

项目	对照组(n=19)	新诊断组(n=37)	非新诊断组(n=27)	统计量	P
CCL3(x±s)	179.7±22.5	302.9±54.11)	301.4±46.41)	F=37.58	<0.001
CCL4[M(P₂₅~P₇₅)]	190.5(179.2~216.7)	302.1(256.0~347.1)¹⁾	298.7(275.9~327.2)¹⁾	H=38.45	<0.001

项目	对照组(n=19)	治疗前组(n=29)	治疗后组(n=35)	F值	P
CCL3	179.7±22.5	346.3±26.5¹⁾	265.8±33.7¹⁾²⁾	155.70	<0.001
CCL4	129.5±24.7	399.8±78.0¹⁾	201.5±40.5¹⁾²⁾	169.74	<0.001

项目	新诊断组(n=37)		t值	P	非新诊断组(n=27)		t值	P
项目	治疗前组(n=19)	治疗后组(n=18)	t值	P	治疗前组(n=10)	治疗后组(n=17)	t值	P
CCL3	345.6±28.1	257.8±34.3	8.54	<0.001	347.7±24.7	274.2±32.0	6.23	<0.001
CCL4	394.7±83.7	191.7±38.7	9.39	<0.001	409.4±69.2	211.9±41.0	9.37	<0.001

项目	0分(n=4)	1分(n=29)	2分(n=23)	3分(n=8)	F值	P
CCL3	231.8±48.8	275.8±34.5¹⁾	325.3±38.7²⁾	367.3±16.0³⁾	22.95	<0.001
CCL4	169.6±44.8	219.2±61.9¹⁾	344.2±99.0²⁾	461.4±45.1³⁾	29.53	<0.001