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摘要：
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摘要：目的 探讨胶原β（1-O）半乳糖基转移酶 [collagen β(1-O) galactosyltransferase 1，
COLGALT1/GLT25D1] 基因在刀豆蛋白A（concanavalin A，Con A）诱导的自身免疫
性肝损伤中的作用及可能机制。方法 6～8周雌性野生型（wild type，WT）小鼠和
Glt25d1基因敲低杂合子（Glt25d1+/-）小鼠分别分为2组（对照组和Con A造模组），
每组中WT小鼠和Glt25d1+/-小鼠均各有6只。Con A以10 mg/kg体质量的剂量经内眦静
脉丛给药。于造模后12 h处死小鼠。留取血浆，检测血清丙氨酸氨基转移酶（alanine
aminotransferase，ALT）和天门冬氨酸氨基转移酶（aspartate transaminase，AST）水
平，留取肝组织，用于肝组织病理学评估和肝脏GLT25D1蛋白表达量测定。制备肝脏、
脾脏及外周血单细胞悬液，运用流式细胞术比较两种小鼠的调节性T（regulatory T，
Treg）细胞、自然杀伤T（natural killer T，NKT）细胞比例，并比较NKT细胞表面凋亡
相关因子配体（factor related apoptosis ligand，FasL）表达差异。结果 对照组和Con A造模组
中Glt25d1+/-小鼠肝组织GLT25D1表达量均显著低于WT小鼠（对照组：0.342 ± 0.168 vs
1.144 ± 0.169，t = 5.841，P = 0.004；Con A造模组：0.264 ± 0.087 vs 0.964 ± 0.058，
t = 11.640，P = 0.0003）。经Con A诱导12 h后，Glt25d1+/-小鼠ALT水平显著高于WT小鼠
[（10155.2 ± 4489.5）U/L vs （3078.5 ± 111.0）U/L；t = -3.522，P = 0.024]。肝组织HE染色
显示Glt25d1+/-小鼠肝板结构损坏更为严重，肝脏坏死区域更加广泛，汇管区炎性细胞浸
润更加显著。流式细胞术检测显示，Con A造模组中Glt25d1+/-小鼠脾脏中Treg细胞频数较
WT小鼠显著减少 [（7.849 ± 1.116）% vs （9.892 ± 1.762）%；t = 2.978，P = 0.008]，肝脏
NKT细胞频数 [（9.244 ± 6.898）% vs （3.376 ± 4.794）%；t = -2.253，P = 0.037] 和NKT细
胞FasL表达量 [（29.62 ± 4.960）% vs （16.43 ± 3.964）%；t = -5.027，P = 0.001] 显著高于
WT小鼠。结论 Glt25d1基因敲低加重Con A诱导的免疫性肝损伤，其机制可能与其降低
Treg细胞频数、增加NKT细胞频数及增强NKT细胞功能有关。
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Abstract: Objective To investigate the role of Collagen β (1-O) galactosyltransferase 1
(COLGALT1/GLT25D1) on concanavalin A (Con A) induced autoimmune liver injury and its
potential mechanism. Methods 6～8 weeks female wild type (WT) mice and Glt25d1 gene
knockdown heterozygous (Glt25d1+/-) mice were randomly divided into two groups (control group
and Con A administration group), respectively, six WT and Glt25d1+/- mice were included in each
group, respectively. Con A was administered via the internal iliac venous plexus at a dose of 10 mg/kg body
mass. Mice were sacrificed after Con A challenged for 12 h. Plasma were collected to detect
the level of serum alanine aminotransferase (ALT) and aspartate transaminase (AST). Liver
tissues were collected to assess the liver histopathology and detect the expression level of
GLT25D1 protein. Liver, spleen and peripheral blood single-cell suspension were prepared
to compare the percentage of regulatory T (Treg) cells and natural killer T (NKT) cells from
different mice, as well as the level of factor related apoptosis ligand (FasL) expressed in NKT
cell surface. Results In both groups, the expression level of GLT25D1 in liver of Glt251d1+/-
mice was significantly lower than that in WT mice, respectively (control group: 0.342 ± 0.168
vs 1.144 ± 0.169, t = 5.841, P = 0.004; Con A induced group: 0.264 ± 0.087 vs 0.964 ± 0.058,
t = 11.640, P = 0.0003). After Con A challenged for 12 h, the serum ALT level of Glt251d1+/-
mice was significantly higher than that of WT mice [(10155.2 ± 4489.5) U/L vs (3078.5 ± 111.0) U/L;
t = -3.522, P = 0.024]. The results of liver tissues after HE staining showed that compared
with WT mice, Glt251d1+/- mice exhibited more serious liver plate structure damage, more
extensive the liver necrosis area and more significant infiltration of inflammatory cells in the
portal area. The results of flow cytometry showed that compared with WT mice, the Treg cells
in spleen of Glt25d1+/- mice in Con A induced group decreased significantly [(7.849 ± 1.116)%
vs (9.892 ± 1.762)%; t = 2.978, P = 0.008], NKT cells [(9.244 ± 6.898)% vs (3.376 ± 4.794)%;
t = -2.253, P = 0.037] and the expression of FasL [(29.62 ± 4.960)% vs (16.43 ± 3.964)%; t = -5.027,
P = 0.001] in liver increased significantly. Conclusions Glt25d1 gene knockdown aggravated Con
A-induced immune liver injury, and its mechanism may be related to the decrease of Treg cells
frequency, the increased frequency of NKT cells and the enhancement of the NKT cell function.
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