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摘要：
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摘要：目的 探讨功能未知基因C6orf120缺失对刀豆蛋白A（concanavalin A，ConA）诱导的自身免疫性
肝炎（autoimmune hepatitis，AIH）大鼠中性粒细胞和巨噬细胞的影响。方法 抽取野生型（wild type，
WT）大鼠和C6orf120基因敲除型（C6orf120-/-）大鼠各24只，随机分成未经ConA诱导组、ConA诱导12 h
组和ConA诱导24 h组共3组，每组中WT大鼠和C6orf120-/-大鼠各8只。其中ConA诱导12 h组和ConA诱
导24 h组大鼠以ConA 35 mg/kg舌下静脉注射建立AIH大鼠模型，于ConA诱导12 h和24 h后分别处死。
检测大鼠血浆丙氨酸氨基转移酶（alanine aminotransferase，ALT）和天门冬氨酸氨基转移酶（aspartate
aminotransferase，AST）水平；采用流式细胞术检测大鼠外周血、肝、脾、肠系膜淋巴结中中性粒细
胞和巨噬细胞的比例；采用全自动五分类血球分析仪检测大鼠全血中性粒细胞比例。结果 未经ConA诱
导组WT大鼠与C6orf120-/-大鼠血浆ALT [（57.03 ± 16.39）U/L vs （55.82 ± 12.35）U/L]和AST [（99.94 ±
18.56）U/L vs（110.86 ± 21.84）U/L]水平差异无统计学意义（t = 0.244、-1.616，P = 0.809、0.115）。
ConA诱导12 h组WT大鼠ALT [（494.27 ± 250.41）U/L vs（206.44 ± 98.66）U/L]和AST [（2043.42 ±
1618.09）U/L vs（777.22 ± 383.95）U/L]水平均显著高于C6orf120-/-大鼠，差异有统计学意义（t =
4.445、3.216，P = 0.001、0.016）。ConA诱导24 h组WT大鼠与C6orf120-/-大鼠血浆ALT [（196.31 ±
76.10）U/L vs（211.63 ± 83.28）U/L]和AST [（892.88.42 ± 354.95）U/L vs（931.44 ± 397.92）U/L]水
平差异无统计学意义（t = -0.384、-0.205，P = 0.707、0.841）。与未经ConA诱导组相比，ConA诱导
12 h和24 h组WT大鼠和C6orf120-/-大鼠血浆ALT与AST水平均显著升高（P均≤ 0.001）；与ConA诱导
12 h组相比，ConA诱导24 h组WT大鼠血浆ALT与AST水平均显著降低（P均≤ 0.001），C6orf120-/-大鼠
血浆ALT与AST水平差异均无统计学意义（P均＞ 0.05）。未经ConA诱导组、ConA诱导12 h组和ConA
诱导24 h组C6orf120-/-大鼠全血中性粒细胞百分比均显著高于WT大鼠[未经ConA诱导组：（21.57 ± 3.88）% vs
（10.63 ± 3.34）%；ConA诱导12 h组：（69.26 ± 2.36）% vs（55.80 ± 6.34）%；ConA诱导24 h组：
（24.57 ± 3.76）% vs（13.09 ± 3.45）%]，差异有统计学意义（t = -6.042、-4.136，-5.513，P均≤ 0.001）。
未经ConA诱导组C6orf120-/-大鼠外周血[（33.24 ± 9.80）% vs（13.77 ± 5.33）%]和脾脏[（8.96 ± 2.65）% vs
（4.32 ± 0.92）%]中性粒细胞比例均显著高于WT大鼠（t = -6.049、-5.941，P均＜ 0.001）；肝脏[（20.85 ±
5.74）% vs（19.25 ± 4.93）%]和肠系膜淋巴结[（0.95 ± 0.26）% vs（0.88 ± 0.45）%]中性粒细胞比例
差异无统计学意义（t = -0.701、-0.466，P = 0.491、0.646）。ConA诱导12 h组和24 h组C6orf120-/-大鼠
外周血[12 h：（59.16 ± 11.72）% vs（39.31 ± 5.88）%；24 h：（50.16 ± 5.95）% vs（38.99 ± 5.91）%]
和肝脏[12 h：（33.43 ± 6.47）% vs（25.20 ± 3.92）%；24 h：（23.23 ± 4.36）% vs（14.34 ± 2.30）%]
中性粒细胞比例均显著高于WT大鼠（P均＜ 0.05），肠系膜淋巴结[12 h：（0.33 ± 0.10）% vs（0.45 ±
0.20）%；24 h：（0.53 ± 0.13）% vs（0.43 ± 0.15）%]和脾脏[12 h：（11.39 ± 4.47）% vs（8.55 ±
5.07）%；24 h：（12.22 ± 2.11）% vs（8.69 ± 5.13）%]中性粒细胞比例差异无统计学意义（P均＞
0.05）。3组间WT大鼠和C6orf120-/-大鼠外周血、肝脏、肠系膜淋巴结及脾脏中性粒细胞比例差异均
有统计学意义（P均＜ 0.001）。未经ConA诱导组C6orf120-/-大鼠外周血[（2.21 ± 0.78）% vs（4.02 ±
1.23）%]、肝脏[（3.18 ± 0.62）% vs（6.72 ± 1.37）%]、肠系膜淋巴结[（4.28 ± 1.25）% vs（7.18 ±
2.21）%]和脾脏[（5.42 ± 0.55）% vs（10.19 ± 1.35）%]巨噬细胞比例均显著低于WT大鼠（t值分别为
4.291、7.797、3.919、11.388，P均≤ 0.001）。ConA诱导12 h和24 h组C6orf120-/-大鼠肠系膜淋巴结中
巨噬细胞比例[12 h：（34.45 ± 8.05）% vs（14.13 ± 6.18）%；24 h：（19.95 ± 6.44）% vs（11.35 ± 1.96）%]
显著高于WT大鼠（t = -5.664，P ＜ 0.001；t = 4.222，P = 0.001），外周血[12 h：（17.63 ± 3.69）% vs
（22.55 ± 4.73）%；24 h：（21.08 ± 5.73）% vs（18.28 ± 7.84）%]、肝脏[12 h：（24.81 ± 8.06）% vs
（25.88 ± 9.91）%；24 h：（9.11 ± 2.84）% vs（9.98 ± 2.55）%]和脾脏[12 h：（15.56 ± 8.16）% vs
（13.89 ± 6.23）%；24 h：（18.39 ± 4.08）% vs（18.70 ± 1.88）%]巨噬细胞比例差异无统计学意义（P
均＞ 0.05）。3组间WT大鼠和C6orf120-/-大鼠外周血、肝脏、肠系膜淋巴结及脾脏巨噬细胞比例差异
均有统计学意义（P均＜ 0.001）。结论 C6orf120基因敲除可促进中性粒细胞增殖而抑制巨噬细胞增
殖。在ConA诱导的大鼠AIH中，C6orf120基因缺失对肝损伤具有保护作用，其机制可能与中性粒细胞
和巨噬细胞的免疫调节作用有关。
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Abstract: Objective To investigate the effects of C6orf120 gene knockout on neutrophils and macrophages
of autoimmune hepatitis (AIH) induced by concanavalin A (ConA) in rats. Methods A total of 24 wildtype (WT) rats and 24 C6orf120-/- knockout (C6orf120-/-) rats were randomly divided into 3 groups: without ConA
administrated group, ConA administrated 12 h group and ConA administrated 24 h group, respectively, eight WT
rats and eight C6orf120-/- rats were allocated into each group. AIH model was established with ConA 35 mg/kg
intravenous injection into rats in ConA administrated 12 h and 24 h groups and rats were sacrificed at 12 h
and 24 h after ConA challenging respectively. The levels of alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) were detected. The ratios of neutrophil and macrophage in peripheral blood, liver,
spleen and mesenteric lymph nodes of rats were determined by flow cytometry. The proportion of neutrophil
in whole blood of rats was detected by automatic five classification hematology analyzer. Results There were
no significant differences in plasma ALT [(57.03 ± 16.39) U/L vs (55.82 ± 12.35) U/L] and AST [(99.94 ±
18.56) U/L vs (110.86 ± 21.84) U/L] levels between WT rats and C6orf120-/- rats of without ConA administrated
group (t = 0.244, -1.616; P = 0.809, 0.115). The ALT [(494.27 ± 250.41) U/L vs (206.44 ± 98.66) U/L] and AST
[(2043.42 ± 1618.09) U/L vs (777.22 ± 383.95) U/L] levels of WT rats were significantly higher than those
of C6orf120-/- rats of ConA administrated 12 h group, and the differences were statistically significant (t =
4.445, 3.216; P = 0.001, 0.016). There were no significant differences in plasma ALT [(196.31 ± 76.10) U/L
vs (211.63 ± 83.28) U/L] and AST [(892.88.42 ± 354.95) U/L vs (931.44 ± 397.92) U/L] levels between WT
rats and C6orf120-/- rats of ConA administrated 24 h group (t = -0.384, -0.205; P = 0.707, 0.841). Compared
with those of without ConA administrated group, the plasma ALT and AST levels of WT rats and C6orf120-/-
rats of ConA administrated 12 h group and ConA administrated 24 h group increased significantly (all P ≤
0.001). Compared with those of ConA administrated 12 h group, the plasma ALT and AST levels of WT
rats of ConA administrated 24 h group decreased significantly (all P ≤ 0.001), and there were no significant
differences in plasma ALT and AST levels of C6orf120-/- rats (all P ＞ 0.05). The percentage of neutrophils in
whole blood of C6orf120-/- rats were significantly higher than those of WT rats of without ConA administrated
group [(21.57 ± 3.88)% vs (10.63 ± 3.34)%], ConA administrated 12 h group [(69.26 ± 2.36)% vs (55.80 ± 6.34)%]
and ConA administrated 24 h group [(24.57 ± 3.76)% vs (13.09 ± 3.45)%], respectively, the differences were
statistically significant (t = -6.042, -4.136, -5.513; all P ≤ 0.001). The neutrophils proportion in peripheral blood
[(33.24 ± 9.80)% vs (13.77 ± 5.33)%] and spleen [(8.96 ± 2.65)% vs (4.32 ± 0.92)%] of C6orf120-/- rats of
without ConA administrated group were significantly higher than those of WT rats (t = -6.049, -5.941; all P ＜
0.001), and there were no significant differences in the neutrophils proportion in liver [(20.85 ± 5.74)% vs
(19.25 ± 4.93)%] and mesenteric lymph nodes [(0.95 ± 0.26)% vs (0.88 ± 0.45)%; t = -0.701, -0.466; P =
0.491, 0.646]. The neutrophils proportion in peripheral blood [12 h: (59.16 ± 11.72)% vs (39.31 ± 5.88)%;
24 h: (50.16 ± 5.95)% vs (38.99 ± 5.91)%] and liver [12 h: (33.43 ± 6.47)% vs (25.20 ± 3.92)%; 24 h: (23.23 ±
4.36)% vs (14.34 ± 2.30)%] of C6orf120-/- rats in ConA administrated 12 h group and ConA administrated
24 h group were significantly higher than those of WT rats, and there were no significant differences in the
proportion of neutrophils in mesenteric lymph nodes [12 h: (0.33 ± 0.10)% vs (0.45 ± 0.20)%; 24 h: (0.53 ±
0.13)% vs (0.43 ± 0.15)%] and spleen [12 h: (11.39 ± 4.47)% vs (8.55 ± 5.07)%; 24 h: (12.22 ± 2.11)% vs
(8.69 ± 5.13)%; all P ＞ 0.05]. The neutrophils proportion in peripheral blood, liver, mesenteric lymph nodes
and spleen of WT rats and C6orf120-/-rats among without ConA administrated group, ConA administrated 12 h
group and ConA administrated 24 h group were statistically significant (all P ＜ 0.001). The macrophage
proportion in peripheral blood [(2.21 ± 0.78)% vs (4.02 ± 1.23)%], liver [(3.18 ± 0.62)% vs (6.72 ± 1.37)%],
mesenteric lymph nodes [(4.28 ± 1.25)% vs (7.18 ± 2.21)%] and spleen [(5.42 ± 0.55)% vs (10.19 ± 1.35)%]
of C6orf120-/- rats of without ConA administrated group were significantly lower than those of WT rats
(t = 4.291, 7.797, 3.919, 11.388; all P ≤ 0.001). The macrophage proportion in mesenteric lymph nodes
[12 h: (34.45 ± 8.05)% vs (14.13 ± 6.18)%; 24 h: (19.95 ± 6.44)% vs (11.35 ± 1.96)%] of C6orf120-/- rats of
ConA administrated 12 h group and ConA administrated 24 h group were significantly higher than those
of WT rats (t = -5.664, P ＜ 0.001; t = 4.222, P = 0.001), and there were no significant differences in the
proportion of macrophage in peripheral blood [12 h: (17.63 ± 3.69)% vs (22.55 ± 4.73)%; 24 h: (21.08 ± 5.73)%
vs (18.28 ± 7.84)%], liver [12 h: (24.81 ± 8.06)% vs (25.88 ± 9.91)%; 24 h: (9.11 ± 2.84)% vs (9.98 ± 2.55)%]
and spleen [12 h: (15.56 ± 8.16)% vs (13.89 ± 6.23)%; 24 h: (18.39 ± 4.08)% vs (18.70 ± 1.88)%; all P ＞
0.05]. The macrophage proportion in peripheral blood, liver, mesenteric lymph nodes and spleen of WT rats
and C6orf120-/- rats among without ConA administrated group, ConA administrated 12 h group and ConA
administrated 24 h group were statistically significant (all P ＜ 0.001). Conclusions Knockout of C6orf120
gene can promote the proliferation of neutrophils and inhibit the proliferation of macrophages. In ConAinduced AIH rats, C6orf120 gene deletion has protective effect on liver injury, which may be related to the
regulation of neutrophils and macrophages.
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