丙型肝炎病毒调控M2各亚型巨噬细胞分化的研究

Regulation of hepatitis C virus on differentiation of subtypes of M2 macrophages

目的探讨丙型肝炎病毒(HCV)对替代性活化的巨噬细胞(M2)各亚型巨噬细胞分化的调控。方法选取2020年1月至2020年12月就诊于山东第一医科大学附属消化病医院的24例慢性HCV患者和同时期来体检的24例健康者,分别作为丙肝组和健康组。采集、分离和纯化外周血单核细胞,诱导其分化为M2a、M2b、M2c型巨噬细胞。比较两组人群M2各亚型巨噬细胞分化情况。结果(1)丙肝组谷丙转氨酶(30.46±1.03)U/L、HCV-RNA(2.6×10^(6)±2.2×10^(6))IU/mL,均高于健康组[(25.84±1.32)U/L、<50 IU/mL],差异有统计学意义(P<0.05)。(2)丙肝组M2a细胞CD 209(94.60±2.91)、CD 86(92.48±7.95)的平均荧光强度低于健康组(282.80±9.59)、(134.50±8.27),丙肝组M2b细胞CD 86(198.00±10.82)、CD 163(216.30±31.69)的平均荧光强度低于健康组[(276.50±9.26)、(338.00±13.13)],丙肝组M2c细胞CD 163的平均荧光强度(447.00±44.65)低于健康组(1458±56.10),差异有统计学意义(P<0.05)。(3)丙肝组M2a细胞缺氧诱导分化因子1(Fizz1)(0.58±0.05)、鞘氨醇激酶1(SPHK1)(0.75±0.09)的基因水平低于健康组[(1.00±0.03)、(1.00±0.06)],丙肝组M2b细胞TNF-α的基因水平(2.53±0.19)低于健康组(4.17±0.35),丙肝组M2c细胞SPHK1的基因水平(1.54±0.11)低于健康组(2.58±0.18)。(4)丙肝组M2a细胞分泌的白细胞介素1受体拮抗物(IL-1ra)[(573.30±41.13)pg/mL]低于健康组[(1493.00±65.38)pg/mL],丙肝组M2b细胞分泌的肿瘤坏死因子-α(TNF-α)[(784.50±32.07)pg/mL]低于健康组[(2370.00±157.20)pg/mL],丙肝组M2c细胞分泌的转化生长因子-β(TGF-β)[(562.60±48.92)pg/mL]低于健康组[(1839.00±70.17)pg/mL],差异有统计学意义(P<0.05)。结论HCV通过抑制M2各亚型巨噬细胞的分化,抑制机体免疫反应。

Objective To investigate the regulation of hepatitis C virus on the differentiation of subtypes of M2 macrophages.Methods A total of 24 chronic hepatitis C patients and 24 healthy subjects who received physical examination at the Affiliated Hospital of Gastroenterology of Shandong the First Medical University from January 2020 to December 2020 were selected as the hepatitis C group and healthy group, respectively. M2 a, M2 b and M2 c macrophages were induced to differentiate from peripheral blood monocytes collected from peripheral venous blood. The differentiation of the three subtypes of M2 macrophages were compared between the two groups.Results(1)The levels of alanine aminotransferase(30.46±1.03) U/L and HCV-RNA(2.6×10;±2.2×10;) IU/mL in the hepatitis C group were higher than those of the healthy group, and the differences were statistically significant [(25.84±1.32) U/L,(<50) IU/mL]. The difference was statistically significant(P<0.05).(2)The expression of CD 209(94.60±2.91) and CD 86(92.48±7.95) on the surface of M2 a subtype macrophages in the hepatitis C group was lower than that of the healthy group [(282.80±9.59),(134.50±8.27)]. The expression of CD 86(198.00±10.82) and CD 163(216.30±31.69) in the M2 b subtype macrophages in the hepatitis C group was lower than that of the healthy group [(276.50±9.26),(338.00±13.13)]. The expression of CD 163(447.00±44.65) in the M2 c subtype macrophages in the hepatitis C group was lower than that of the healthy group(1 458±56.10). The difference was statistically significant(P<0.05).(3)The expression of Fizz1(0.58±0.05) and SPHK1(0.75±0.09) in subtype M2 a macrophages of hepatitis C group was lower than that of healthy group [(1.00±0.03),(1.00±0.06)]. The expression of TNF-α(2.53±0.19) in subtype M2 b macrophages of hepatitis C group was lower than that of healthy group(4.17±0.35). And the expression of SPHK1(1.54±0.11) in subtype M2 c macrophages of hepatitis C group was lower than that of healthy group(2.58±0.18). The difference is statistically significant(P<0.05).(4)The secretion of IL-1 ra(573.30±41.13) pg/mL in M2 a subtype macrophages in hepatitis C group was lower than that of healthy group(1 493.00±65.38) pg/mL. The secretion of TNF-α(784.50±32.07) pg/mL in M2 b subtype macrophages in hepatitis C group was lower than that in healthy group(2 370.00±157.20) pg/mL. And the secretion of TGF-β(562.60±48.92) pg/mL in M2 c subtype macrophages in hepatitis C group was lower than that of healthy group(1 839.00±70.17) pg/mL. The difference was statistically significant(P<0.05).Conclusion HCV can inhibit the differentiation of M2 subtypes of macrophages to inhibit the immune response of the body.