C6orf120基因敲除对刀豆蛋白A诱导的自身免疫性肝炎大鼠中性粒细胞和巨噬细胞的影响

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...

Effects of C6orf120 gene knockout on neutrophils and macrophages of autoimmune hepatitis induced by concanavalin A in rats

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.