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摘要：
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摘要：目的 初步探索糖基转移酶Colgalt2基因敲除对刀豆蛋白A（concanavalin A，Con
A）诱导的自身免疫性肝炎（autoimmune hepatitis，AIH）小鼠模型中CD4 T细胞记忆
和归巢的影响。方法 随机抽取无特定病原体（specific pathogen free，SPF）级6～8周的
基因敲除（Colgalt2-/-）小鼠与野生型Colgalt2+/+小鼠各18只，建立Con A诱导的AIH小鼠
模型。检测小鼠血浆中丙氨酸氨基转移酶（alanine aminotransferase，ALT）和天门冬氨
酸氨基转移酶（aspartate aminotransferase，AST）水平，采用HE染色观察肝组织病理
变化，采用体外磁珠分选脾脏CD4+
T细胞，采用流式细胞术检测小鼠外周血、脾脏及
分选后CD4+
T细胞亚型比例。结果 与Colgalt2+/+小鼠相比，Con A刺激12 h Colgalt2-/-小
鼠血清ALT [（15610 ± 2869）U/L vs（5009 ± 1042）U/L；t = 3.474，P = 0.006] 和AST
[（16080 ± 2631）U/L vs（4453 ± 893.7）U/L；t = 4.185，P = 0.002] 水平均显著升高。
Con A刺激12 h Colgalt2-/-小鼠肝损伤分级显著高于Colgalt2+/+小鼠 [（3.17 ± 0.17）级
vs（2.17 ± 0.17）级]，差异有统计学意义（t = 4.243，P = 0.002）。Con A刺激12 h
Colgalt2-/-小鼠脾脏CD4+
CD28-
[（97.80 ± 0.46）% vs（88.85 ± 3.35）%；t = 2.650，P =
0.024] 和CD4+
CD62L-
[（97.98 ± 0.46）% vs（88.68 ± 3.38）%；t = 2.669，P = 0.023] 均
显著高于Colgalt2+/+小鼠。体外脾脏实验表明，Con A刺激12 h Colgalt2-/-小鼠CD4+
CD28-
[（94.88 ± 1.20）% vs（82.97 ± 2.70）%；t = 4.295，P = 0.002] 和CD4+
CD62L-
[（99.95 ±
0.02）% vs（99.24 ± 0.26）%；t = 2.731，P = 0.021] 均显著高于Colgalt2+/+小鼠。结论
Colgalt2基因敲除使脾脏CD4+
CD28-
和CD4+
CD62L细胞亚群表达上调从而加重Con A诱
导的AIH。
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Abstract: Objective To investigate the effects of glycosyltransferase Colgalt2 gene knockout
on homing and memory of CD4 T cells in concanavalin A (Con A) induced autoimmune
hepatitis (AIH) mice. Methods The specific pathogen free (SPF) Colgalt2-/- mice (18 cases)
and Colgalt2+/+ (wild type) mice (18 cases) were randomly selected and the mice were
6～8 weeks. The model of AIH induced by Con A was established. Serum alanine
aminotransferase (ALT) and aspartate aminotransferase (AST) levels of mice were detected
and histopathological change of liver tissue was observed by HE staining. The magnetic bead
sorting technique was used to isolate the CD4+ T cells of spleen. CD4+ subsets in peripheral
blood mononuclear cells and splenocytes were analyzed by flow cytometry. Results Compared
with those of Colgalt2+/+ mice, the ALT [(15610 ± 2869) U/L vs (5009 ± 1042) U/L; t = 3.474,
P = 0.006] and AST [(16080 ± 2631) U/L vs (4453 ± 893.7) U/L; t = 4.185, P = 0.002] levels
of Colgalt2-/- mice increased significantly after Con A administrated for 12 h. The liver injury
grade of Colgalt2-/- mice was higher than that of Colgalt2+/+ mice after Con A administrated
for 12 h [(3.17 ± 0.17) grade vs (2.17 ± 0.17) grade], the difference was statistically
significant (t = 4.243, P = 0.002). Compared with those of Colgalt2+/+ mice, the percentage
of CD4+
CD28-
[(97.80 ± 0.46)% vs (88.85 ± 3.35)%; t = 2.650, P = 0.024] and CD4+
CD62L-
[(97.98 ± 0.46)% vs (88.68 ± 3.38)%; t = 2.669, P = 0.023] increased significantly in
Colgalt2-/- mice after Con A administrated for 12 h in spleen. In vitro, compared with those
of Colgalt2+/+ mice, the percentage of CD4+
CD28-
[(94.88 ± 1.20)% vs (82.97 ± 2.70)%; t =
4.295, P = 0.002] and CD4+
CD62L-
[(99.95 ± 0.02)% vs (99.24 ± 0.26)%; t = 2.731, P = 0.021]
also increased significantly in Colgalt2-/- mice after Con A administrated for 12 h in spleen.
Conclusions Colgalt2 gene knockout aggravated Con A induced AIH in mice by increasing
the expression of CD4+
CD28-
and CD4+
CD62L-
T subsets in spleen.
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