MAPK通路主要激酶抑制剂对亚洲带绦虫感染乳猪肝细胞生长抑制率和凋亡率的影响

目的 了解MAPK特异性抑制剂干预后亚洲带绦虫感染乳猪肝细胞MAPK信号通路相关基因表达、细胞活力和细胞凋亡的变化。方法 采集亚洲带绦虫成虫标本并收集虫卵制作悬液。将乳猪随机分为感染组、三种抑制剂干预组和正常对照组,感染组和抑制剂干预组均以定量虫卵15万个/头灌胃感染,灌胃后将抑制剂干预组乳猪分别腹腔注射U0126、SB203580和SP600125 1 mg/kg,隔天1次,连续7次。感染后第15 d麻醉处死各组乳猪获取肝脏。采用cck-8法和流式细胞仪分别检测肝细胞生长抑制率和肝细胞凋亡率,并用RT-PCR技术检测肝细胞ERK1、p38和JNK1基因mRNA的相对表达量。结果 感染组和U0126、SB203580、SP600125干预组肝细胞生长抑制率分别为(23.1±1.26)%、(12.59±1.2)%、(16.48±0.7)%、(18.3±0.75)%,U0126组与感染组相比(t=9.223,P<0.05)、SB203580组与感染组相比为(t=11.532,P<0.05)、SP600125组与感染组相比(t=6.775,P<0.05),各干预组均较感染组的生长抑制率明显降低。与感染组相比,U0126干预组的肝细胞凋亡率与感染组相比增高 (t=-5.934,P<0.001),SB203580干预组与感染组相比则降低 (t=3.821,P<0.01),SP600125干预组与感染组相比(t=-1.545,P＞0.05)无显著性差异。与感染组相比,U0126干预组(t=3.462,P<0.05)ERK1 mRNA表达量下调;SB203580干预组(t=3.267,P<0.05) p38 mRNA表达量下调;SP600125干预组(t=3.363,P<0.001) JNK1 mRNA的表达量下调。结论 亚洲带绦虫感染乳猪后,MAPK特异性抑制剂可以通过影响乳猪肝细胞MAPK相关基因的mRNA水平减轻肝细胞的生长抑制和凋亡。

Major kinase inhibitors in the MAPK pathway effects on the growth inhibition rate and apoptosis rate of hepatocytes in the porkets infected by Taenia asiatica

To understand the changes of MAPK signaling pathway-related gene expression, cell viability and cell apoptosis in porkets hepatocytes infected with Taenia asiatica after MAPK-specific inhibitor intervention. Specimens of adult Taenia asiatica were collected and egg suspension was produced. The porkets were randomly divided into the infection groups, the three inhibitor intervention groups and the normal control group. The infection group and the inhibitor intervention group were orally administrated with 1.5×10⁶ eggs per porket at Day 0 post-infection. In the inhibitor intervention group, each porket was intraperitoneally injected with inhibitors U0126, SB203580 and SP600125 1 mg/kg from the day of intragastric administration, once every other day for 7 consecutive injections. Livers were obtained by anesthetizing the groups of porkets on the 15th day after infection. CCK-8 method and flow cytometry were used to detect the growth inhibition rate and apoptotic rate of hepatocytes in porkets. The relative expression of ERK1, p38 and JNK1 genes in hepatocytes of porkets was detected by real-time fluorescence quantitative PCR. The inhibition rates of hepatocyte growth in infection group and U0126, SB203580 and SP600125 intervention group were (23.11±1.26)%, (12.59±1.2)%, (16.48±0.7)%, (18.3±0.75)%. The inhibition rates of hepatocyte growth in intervention group were significantly lower than those in infection group (t=9.223,P<0.05). Compared with the infected group, the apoptosis rates of hepatocytes in and the U0126 intervention group wes significantly higher (t=-5.934,P<0.001),but decreased in SB203580 intervention group (t=3.821,P<0.01), and there was no significant difference in the SP600125 intervention group (t=-1.545,P>0.05). Compared with the infected group, the expression of ERK1 gene in U0126 intervention group was significantly down-regulated (t=3.462,P<0.05); the expression of p38 gene in SB203580 intervention group was significantly down-regulated (t=3.267,P<0.05); the expression of JNK1 gene in SP600125 intervention group was significantly down-regulated (t=3.363,P<0.001). After the Taenia asiatica infected porkets, MAPK-specific inhibitors can alleviate hepatocyte growth inhibition and apoptosis by affecting the mRNA level of MAPK-related genes in porkets hepatocytes.