Pro-Inflammatory and Anti-Inflammatory Cytokines Levels are Significantly Altered in Cerebrospinal Fluid of Unruptured Intracranial Aneurysm (UIA) Patients

Figures & data

Table 1 Clinical Characteristics and Basic Laboratory Parameters in UIA Patients and the Control Group

Clinical Characteristics
Variable	Control Group		UIA Group		p-value
	Mean ± SD	Median (interquartiles)	Mean ± SD	Median (interquartiles)
Age [years]	62 ± 9	61 (56–68)	57 ± 12	59 (53–66)	0.1552^NP
Sex: Female/Male	8/7	-	33/7	-	0.0623^&
Weight [kg]	81 ± 14	82 (70–90)	73 ± 16	69 (63–83)	0.0804^NP
Growth [m]	1.67 ± 0.08	1.68 (1.60–1.72)	1.64 ± 0.05	1.64 (1.62–1.68)	0.2424^NP
BMI [kg/m^2]	28.84 ± 4.30	29.41 (25.69–32.87)	26.99 ± 5.40	25.78 (23.18–30.09)	0.1608^NP
Systolic BP [mmHg]	142 ± 15	143 (127–153)	138 ± 20	137 (123–150)	0.4440^PT
Diastolic BP [mmHg]	84 ± 12	85 (70–95)	81 ± 9	80 (76–90)	0.5939^NP
Heart rate [minute]	74 ± 13	70 (63–83)	71 ± 15	74 (65–78)	0.7864^NP
BASIC LABORATORY PARAMETERS
	Mean ± SD	Median (interquartiles)	Mean ± SD	Median (interquartiles)	p-value
WBC [x10^3/µL]	6.18 ± 1.71	5.97 (4.71–7.52)	7.51 ± 2.18	7.28 (6.30–8.60)	0.0378^PT
RBC [x10^6/µL]	4.56 ± 0.36	4.51 (4.45–4.73)	4.47 ± 0.48	4.45 (4.18–4.77)	0.5416^PT
HGB [g/dl]	13.55 ± 0.91	13.50 (12.90–14.00)	13.47 ± 1.26	13.65 (12.80–14.25)	0.8202^PT
HCT [%]	40.19 ± 2.71	40.20 (37.50–41.40)	41.07 ± 9.37	39.60 (37.50–42.50)	0.8444^NP
PLT [x10^3/µL]	263 ± 53	274 (238–300)	229 ± 66	233 (184–280)	0.0497^NP
MPV [fl]	10.3 ± 0.6	10.3 (9.8–10.7)	11.0 ± 1.0	11.0 (10.3–11.6)	0.0155^PT
P-LCR[%]	33.8 ± 25.3	28.2 (23.2–30.5)	32.9 ± 8.0	33.3 (26.8–37.7)	0.0679^NP
PT [s]	12.8 ± 0.6	12.9 (12.2–13.1)	12.3 ± 0.5	12.2 (11.9 12.6)	0.0012^PT
INR	0.97 ± 0.05	0.99 (0.91–1.02)	0.92 ± 0.05	0.91 (0.88–0.94)	0.0006^PT
APTT [s]	28.1 ± 3.4	27.1 (25.9–29.9)	28.5 ± 2.2	28.5 (27.1–29.9)	0.5544^PT
Fibrinogen [mg/dl]	319 ± 73	300 (266–383)	350 ± 64	337 (298–388)	0.1198^NP
Na^+ [mmol/l]	138 ± 4	139 (136–140)	138 ± 3	138 (137–140)	0.6070^NP
K^+ [mmol/l]	4.7 ± 0.5	4.7 (4.3–5.1)	4.4 ± 0.4	4.4 (4.1–4.7)	0.0335^PT
Glucose [mg/dl]	99 ± 18	99 (88–103)	101 ± 19	95 (88–111)	0.8298^NP
Urea [mg/dl]	35 ± 17	29 (22–49)	32 ± 10	31 (26–41)	0.4114^PT
Creatinine [mg/dl]	0.80 ± 0.14	0.81 (0.70 −0.87)	0.78 ± 0.12	0.79 (0.69–0.85)	0.6235^PT
eGFR [mL/min/1.73 m²]	94 ± 24	98 (77–105)	87 ± 16	89 (76–98)	0.2807^PT

Notes: The bold font of p-value means statistical significance (p<0.05). Conversion factors to SI units are as follows: for WBC – 1.0, for RBC – 1.0, for HGB – 10.0, for PLT – 1.0, for glucose – 0.0555, for creatinine – 88.4. ^&Yates’ chi-square test, ^PTparametric Student’s t-test, ^NPnon-parametric U Mann–Whitney test.

Abbreviations: APTT, activated partial thromboplastin; BMI, body mass index; eGFR, estimated glomerular filtration rate; HCT, hematocrit; HGB, hemoglobin; INR, International Normalized Ratio Time; K⁺, potassium ion; MPV, mean platelet volume; Na⁺, sodium ion; P-LCR, platelet‐large cell ratio; PLT, platelet count; PT, prothrombin time; RBC, red blood cell count; SD, standard deviation; UIA, unruptured intracranial aneurysm; WBC, white blood cell count.

Table 2 Risk Factors of Intracranial Aneurysm Development in the Group of UIA Patients and the Control Group

Risk Factors	Control Group N (%)	UIA Group N (%)	p-value
Smoking tobacco^#	4 (27)	21 (53)	0.0866
Drinking alcohol^@	3 (20)	5 (13)	0.7842^&
Diabetes	4 (27)	3 (8)	0.1484^&
Obesity^$	7 (47)	11 (28)	0.3046^&
Hypertension	7 (47)	28 (70)	0.1091
Cardiovascular diseases	0 (0)	7 (18)	0.2005^&
Head injury	0 (0)	3 (8)	- ^%

Notes: ^&Yates’ chi-square, ^#More than 20 cigarettes a day for over 12 months and more, ^@More often than occasionally, ^$body mass index (BMI) > 30.0 (kg/m²), ^%Too few cases, to perform the statistic test.

Abbreviations: N, number; UIA, unruptured intracranial aneurysm.

Figure 1 (A–D). Localization (A) and geometry (B–D) of aneurysms of UIA patients (n=40).

Abbreviations: AcomA, anterior communicating artery; ICA, internal carotid artery; MCA, middle cerebral artery; mm, millimeter; UIA, unruptured intracranial aneurysm.

Table 3 CSF Concentrations of Interleukins, Cytokines, Chemokines, and Growth Factors in UIA Patients and the Control Group

Variable	Control Group	UIA Group
CSF [pg/mL]	Median (Interquartiles)	Median (Interquartiles)	2-Tailed p-value
IL-1β	0.15 (0.04–0.20)	0.30 (0.18–0.49)	0.0006*
IL-1ra	32.19 (32.19–84.77)	114.96 (39.65–163.43)	0.0165*
IL-2	1.22 (0.85–1.40)	2.04 (1.36–2.49)	0.0004*
IL-4	0.21 (0.13–0.35)	0.54 (0.29–0.84)	0.0014*
IL-5	12.73 (6.53–20.27)	74.51 (29.93–96.85)	<0.0001*
IL-6	2.15 (0.90–3.06)	2.05 (1.61–2.86)	0.5556
IL-7	1.55 (0.90–1.61)	2.13 (1.55–3.62)	0.0037*
IL-8	13.73 (12.11–16.58)	18.79 (14.49–26.75)	0.0133*
IL-9	23.34 (13.53–56.99)	17.21 (11.54–43.08)	0.6070
IL-10	1.14 (0.31–1.94)	1.68 (0.17–2.47)	0.3066
IL-12	0.17 (0.17–1.09)	1.31 (0.17–2.37)	0.0266*
IL-13	0.40 (0.07–1.40)	0.97 (0.30–1.16)	0.0065*
IL-15	#	155.36 (105.34–229.84)^&	-
IL-17	1.54 (0.54–2.62)	1.96 (1.41–3.61)	0.1392
TNF-α	1.94 (1.44–4.04)	4.53 (3.06–8.67)	0.0002*
IFN-γ	4.93 (3.78–5.49)	7.41 (5.96–9.63)	0.0002*
Eotaxin	4.19 (3.40–4.69)	4.88 (3.90–5.65)	0.1977
IP-10	3439.31 (3009.68–4052.98)	3865.06 (2405.82–5007.00)	0.5705
MCP-1	123.93 (108.53–160.50)	169.10 (122.70–213.30)	0.0205*
MIP-1α	2.89 (1.56–4.35)	4.22 (2.16–11.01)	0.0545
MIP-1β	24.68 (11.64–34.93)	34.18 (14.34–71.73)	0.2329
RANTES	72.44 (42.95–186.74)	44.52 (20.80–132.77)	0.3156
GM-CSF	#	0.66 (0.21–0.94)	-
G-CSF	117.70 (62.67–189.48)	156.00 (93.72–389.28)	0.0771
VEGF	10.86 (10.86–51.99)	63.80 (16.70–115.83)	0.0497*
PDGF-BB	24.97 (18.27–28.25)	26.05 (11.29–39.99)	0.4710
bFGF	9.21 (7.23–13.20)	14.06 (8.74–20.65)	0.1109

Notes: ^#Concentration not detected, ^&Concentration detected only in five cases,*And bold of a 2-tailed p-value – the level of significance < 0.05.

Abbreviations: CSF, cerebrospinal fluid; IL, interleukin; TNF-α, tumor necrosis factor-α; INF-γ; interferon-γ; GM-CSF, granulocyte monocyte colony-stimulating factor; G-CSF, granulocyte colony-stimulating factor; IP-10, interferon gamma-induced protein 10; MCP-1, monocyte chemoattractant protein-1; MIP-1-α, macrophage inflammatory protein-1α; MIP-1β, macrophage inflammatory protein-1β; RANTES, regulated on activation, normal T cell expressed and secreted; VEGF, vascular endothelial growth factor; PDGF-BB, platelet-derived growth factor-BB; Bfgf, basic fibroblast growth factor; UIA, unruptured intracranial aneurysm.

Table 4 Serum Concentrations of Interleukins, Cytokines, Chemokines, and Growth Factors in UIA Patients and the Control Group

Variable	Control Group	UIA Group
Serum [pg/mL]	Median (Interquartiles)	Median (Interquartiles)	2-Tailed p-value
IL-1β	1.44 (1.14–1.70)	1.39 (1.19–1.39)	0.3213
IL-1ra	128.76 (128.76–457.24)	128.76 (128.76–394.74)	0.7430
IL-2	0.76 (0.56–1.52)	0.38 (0.38–1.14)	0.7430
IL-4	4.76 (3.64–6.48)	5.49 (4.41–6.67)	0.7430
IL-5	50.92 (50.92–50.92)	#	-
IL-6	0.98 (0.56–6.42)	0.70 (0.70–0.98)	0.9626
IL-7	14.48 (14.48–24.32)	14.48 (10.61–14.48)	0.2766
IL-8	7.30 (4.46–31.40)	6.93 (4.82–8.36)	0.7430
IL-9	631.96 (582.24–684.16)	620.91 (618.92–677.46)	0.5414
IL-10	1.24 (1.24–7.24)	2.37^$	-
IL-12	4.36 (4.36–4.36)	#	-
IL-13	0.66 (0.28–3.92)	#	-
IL-15	#	#	-
IL-17	18.26 (15.96–24.04)	20.28 (17.24–24.29)	0.9026
TNF-α	22.48 (19.08–25.84)	22.95 (21.02–24.40)	0.9626
IFN-γ	12.30 (10.66–14.32)	11.89 (10.23–11.89)	0.2359
Eotaxin	127.44 (65.86–170.30)	123.73 (96.20–166.22)	0.7866
IP-10	1541.98 (1126.02–2028.18)	1451.39 (1069.70–1587.32)	0.7133
MCP-1	26.00 (19.20–41.34)	21.34 (19.65–25.27)	0.3079
MIP-1α	2.22 (1.84–3.22)	2.28 (1.57–2.81)	0.5414
MIP-1β	260.60 (245.34–299.62)	268.80 (263.47–274.52)	0.4807
RANTES	10631.40 (7868.50–13850.88)	12801.02 (11396.66–13736.82)	0.1426
GM-CSF	0.83^&	22.52^$	-
G-CSF	122.76 (104.80–171.74)	133.87 (100.02–162.97)	0.7880
VEGF	#	#	-
PDGF-BB	3127.40 (2583.08–3986.14)	2401.26 (2191.16–3165.48)	0.6441
bFGF	34.94 (33.04–42.06)	33.05 (33.05–36.85)	0.8148

Notes: ^#Concentration not detected, ^$Concentration detected only in two cases, ^&Concentration detected only in one case.

Abbreviations: IL, interleukin; TNF-α, tumor necrosis factor-α; IFN-γ, interferon-γ; GM-CSF, granulocyte monocyte colony-stimulating factor; G-CSF, granulocyte colony-stimulating factor; IP-10, interferon gamma-induced protein 10; MCP-1, monocyte chemoattractant protein-1; MIP-1-α, macrophage inflammatory protein-1α; MIP-1β, macrophage inflammatory protein-1β; RANTES, regulated on activation, normal T cell expressed and secreted; VEGF, vascular endothelial growth factor; PDGF-BB, platelet-derived growth factor-BB; bFGF, basic fibroblast growth factor; UIA, unruptured intracranial aneurysm.

Figure 2 (A–I). CSF IL-1β (A), IL-1ra (B), IL-2 (C), IL-4 (D), IL-5 (E), IL-7 (F), IL-8 (G), IL-12 (H), and IL-13 (I) concentrations in UIA patients (n=40) and the control group (n=15).

Note: *p < 0.05, ** p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.

Abbreviations: C, control group; CSF, cerebrospinal fluid; IL, interleukin; IL-1ra, IL-1 receptor antagonist; UIA, unruptured intracranial aneurysm.

Figure 3 (A–D). CSF TNF-α (A), IFN-γ (B), MCP-1 (C), and VEGF (D) concentrations in UIA patients (n=40) and the control group (n=15).

Note: *p < 0.05, ***p ≤ 0.001.

Abbreviations: C, control group; CSF, cerebrospinal fluid; IFN-γ, interferon-γ; MCP-1, monocyte chemoattractant protein-1; TNF-α, tumor necrosis factor-α; UIA, unruptured intracranial aneurysm; VEGF, vascular endothelial growth factor.

Figure 4 (A–E). CSF IL-2 (A), IL-6 (B), IL-8 (C), MIP-1α (D), and MCP-1 (E) concentrations as compared to the serum values in UIA patients (n=40) and the control group (n=15).

Note: *p < 0.05, ***p ≤ 0.001, ****p ≤ 0.0001.

Abbreviations: CSF, cerebrospinal fluid; MCP-1, monocyte chemoattractant protein-1; MIP-1α, macrophage inflammatory proteins-1α; UIA, unruptured intracranial aneurysm.

Figure 5 CSF IL-1β concentration in UIA patients depending on aneurysm’s location.

Note: *p < 0.05.

Abbreviations: CSF, cerebrospinal fluid; ICA, internal carotid artery; IL-1β, interleukin 1 beta; MCA, middle cerebral artery; UIA, unruptured intracranial aneurysm.

Figure 6 The heat map of the relationship between interleukins, cytokines, chemokines, and growth factors in the CSF of UIA patients (n=40).

Abbreviations: CSF, cerebrospinal fluid, IL, interleukin, TNF-α, tumor necrosis factor-α, INF-γ, Interferon-γ, GM-CSF, granulocyte monocyte colony-stimulating factor, G-CSF, granulocyte colony-stimulating factor, IP-10, Interferon gamma-induced protein 10, MCP-1, monocyte chemoattractant protein-1, MIP-1-α, macrophage inflammatory protein-1α, MIP-1β, macrophage inflammatory protein-1β, RANTES, regulated on activation, normal T cell expressed and secreted, VEGF, vascular endothelial growth factor, PDGF-BB, platelet-derived growth factor-BB, bFGF basic fibroblast growth factor, UIA, unruptured intracranial aneurysm.

Figure 7 (A–F). Correlation coefficient analysis of CSF IL-1β (A), IL-8 (B), TNF-α (C), and IL-13 (D) concentrations with the size of aneurysms and CSF IL-6 (E) and MCP-1 (F) concentrations with the number of aneurysms.

Abbreviations: CSF, cerebrospinal fluid; IL, interleukin; MCP-1, monocyte chemoattractant protein-1; TNF-α, tumor necrosis factor-α.