Analysis Of IL-6 Expression Level In Lymphatic Fluid And Its Influencing Factors

Background and purposeLymphatic malformation is a benign,low-flow malformation of the lymphatic system.The International Society for the Study of Vascular Abnormalities(ISSVA)reclassified lymphatic malformations according to the size of lymphatic channels.The most commonly used diagnostic methods in clinic are imaging examinations,such as color Doppler ultrasound and magnetic resonance imaging.With the development of interventional technology and the emergence of various types of sclerosing agents,fine-needle puncture sclerotherapy has replaced the traditional surgical excision method with its advantages of minimally invasive and rapid effect.Sclerotherapy requires multiple treatments,or targeted drugs can only control the disease and can not achieve the goal of eradication.Against the backdrop of the lack of specific detection indicators such as changes and severity of the disease,new discoveries are needed to advance understanding of the disease.The study found that the formation of lymphatic malformation may be related to cytokines,and the level of Interleukin-6(IL-6)increased significantly after the treatment of injection of OK-432.Is there also an increase in IL-6 level in Lymphatic vessel fluid of untreated Lymphatic vessel malformations?Does intracapsular hemorrhage of lymphatic malformation affect the level of IL-6?Is IL-6 expressed in cell membrane,cytoplasm or nucleus?What are the clinical factors related to the level of IL-6?What is the level of protein and glucose in lymphatic fluid?Will intracapsular hemorrhage affect the protein level?The above questions are problems encountered in clinical work.At present,most of the above questions have not been fully answered by relevant literature.This topic will focus on the above questions.The purpose of this study is to analyze the relationship between IL-6 and clinical factors by detecting the level and change of cytokines IL-6 and biochemical components in lymphatic fluid,and speculate that IL-6 is related to the progression or severity of lymphatic malformation.MethodThe serum and lymphatic fluid of 46 children with lymphatic malformation hospitalized in the Department of Vascular and Interventional Vascular Surgery,Children’s Hospital Affiliated to Shandong University from April 1,2020 to January 1,2021 were prospectively tested for IL-6 cytokines.The pathological tissue samples of 4 children with lymphangioma or lymphangioma diagnosed pathologically after the surgery of Children’s Hospital Affiliated to Shandong University from 2019 to 2022 were tested for IL-6 immunohistochemistry.Retrospective case data of 170 children with lymphangitic malformations hospitalized in the Department of Vascular and Interventional Vascular Surgery of Children’s Hospital Affiliated to Shandong University from November 1,2019 to October 31,2021 were collected to analyze the relationship between IL6 and clinical factors.The data were statistically analyzed using IBM SPSS 21.Through the normality test and variance homogeneity test,the data of continuous variables that do not conform to the normal distribution are expressed by M(P25,P75).The difference between the two groups was expressed by Mann-Whitney U test.Linear correlation analysis is used for bivariate normal quantitative variables,and rank correlation analysis is used for bivariate quantitative or hierarchical variables.One or more variables are skewed or hierarchical variables.P<0.05,which is regarded as statistically significant.Results1.The level of IL-6 in lymphatic fluid was different from that in serum in children with lymphatic malformationThere was a statistically significant difference between IL-6 232.04(7.42-2888.1)pg/mL in lymphatic fluid and IL-6 1.96(1.7-3.11)pg/mL in serum;There was a statistically significant difference between IL-10(Interleukin-10)5.16(3.39-9.31)pg/mL in lymphatic fluid and 2.14(1.72-3.45)pg/mL in serum(P<0.001).But IL-2(Interleukin-2),IL-4(Interleukin-4),TNF a(Tumor necrosis factor-a)And IFN y(Interferon-y)The level difference was not statistically significant.2.There was significant difference in the level of IL-6 in lymphatic fluid between the intracapsular hemorrhage group and the non intracapsular hemorrhage groupThe concentration of IL-6 in the bleeding group was 2538.77(420.18-8261.36)pg/mL,which was significantly higher than that in the non-bleeding group 6.57(4.45-78.21)pg/mL(P<0.001).3.Positive expression of IL-6 was found in pathological tissue samplesPartial IL-6 positive expression was found in the whole visual field of 3 pathological specimens.4.IL-6 levels are associated with the patient’s age,treatment history,local pain symptoms,and bleeding statusThe median level of IL6 in the group with lymphatic malformation during pregnancy or birth was 19.25(5.06,59.00)pg/mL,and the median level of IL6 in the group with later detection was 1177.16(80.67,6968.67)pg/mL.There was a statistical difference in the distribution of total IL6 levels between the two groups(Z=4.759,P<0.001).The median level of IL-6 in the group with treatment history was 2383.54(86.65,1048.02)pg/mL,and the median level of IL-6 in the group without treatment was 205.93(16.40,3914.94)pg/mL.There was a statistical difference between the two groups in the overall level of IL-6(Z=2.699,P=0.007).The median level of IL6 in the group with pain in the focus area was 3980.87(1936.04,17032.89)pg/mL,and the median level of IL6 in the group without pain in the focus area was 205.93(18.84,5156.77)pg/mL.There was a statistical difference between the two groups in the overall level of IL6(Z=3.596,P<0.001).The median level of IL6 in the yellow lymphatic fluid group was 21.17(6.47,130.21)pg/mL,the median level of IL6 in the light red lymphatic fluid group was 96.76(18.81,3555.71)pg/mL,and the median level of IL6 in the dark red lymphatic fluid group was 3250.17(438.57,1011.67)pg/mL.There was a statistical difference in the overall distribution of IL6 levels(H=55.171,P<0.001).There were statistical differences in the correlation between IL6 and age(r=0.217,P=0.04),with pregnancy or birth(r=0.300,P<0.001),with treatment history(r=-0.17,P=0.007),with pain(r=0.227,P<0.001),and with lymphatic fluid color(r=-0.458,P<0.001).ConclusionThe inflammatory reaction mediated by IL-6 in lymph does not trigger the occurrence of cytokine storm in blood.The development of lymphatic malformation and the severity of inflammation are related to IL-6.The inflammatory reaction caused by lymphatic fluid did not affect the content of protein and chlorine in the lymphatic cyst fluid.