摘要目的：分析 CHB 患者体内磷脂酰肌醇3-激酶（PI3K）／蛋白激酶 B（Akt）／哺乳动物雷帕霉素靶（mTOR）信号通路对 Th17分化的影响。方法收集20例 CHB 患者肝组织作为实验组（CHB组），20例肝脏良性腺瘤周围的肝组织作为对照组（对照组）。实时 PCR 检测两组肝组织中 IL-17 mRNA、CD3γmRNA 的表达，Western 印迹检测 Akt、磷酸化-蛋白激酶 B（p-Akt）、p-S6和 IL-17的表达，免疫荧光观察 p-Akt 和 IL-17共定位情况，分析共表达 p-Akt 和 IL-17的 Th17在两组肝组织中的分布，同时行 HBV 相关血清学检查。两组间数据比较采用 t 检验，组间相关性采用 Pearson 直线相关分析。结果在 CHB 组肝组织中，IL-17 mRNA 和 CD3γmRNA 的表达水平分别为22．530±5．657和12．791±3．433，均明显高于对照组的1．025±0．246（t ＝16．99，P ＜0．01）和1．033±0．305（t ＝15．26， P ＜0．01）；IL-17 mRNA／CD3γmRNA 为1．782±0．213，高于对照组的1．014±0．144（t ＝13．49，P ＜0．01）；IL-17蛋白为0．479±0．069，高于对照组的0．037±0．007（t＝18．15，P ＜0．01）。IL-17 mRNA 在CHB 患者肝脏组织中的表达水平与 ALT 呈正相关（r＝0．664，P ＜0．01），与 HBV DNA 载量无相关性（r＝0．053，P ＞0．05）。CHB 组肝组织内的 p-Akt 和 p-S6表达分别为0．355±0．015和0．380±0．043，高于对照组的0．145±0．009（t＝30．93，P ＜0．01）和0．140±0．021（t＝14．18，P ＜0．01）；而两组 Akt 的表达差异无统计学意义（t＝2．10，P ＞0．05）。在 CHB 组肝组织内存在高表达 IL-17和 p-Akt 的双阳性Th17，其双阳性细胞计数为（4．872±0．513）／高倍视野，明显高于对照组的（1．440±0．301）／高倍视野（t＝18．25，P ＜0．01）。结论 CHB 患者肝组织内存在 Th17的激活；PI3K／Akt／mTOR 信号通路的活化可能与 IL-17表达和 Th17激活有关。提示该信号通路的活化和 Th17的激活可能在 CHB 疾病发展中起重要作用。

abstractsObjective To investigate the influence of phosphatidy linositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR)signaling pathway on differentiation of T helper (Th)17 cells in chronic hepatitis B (CHB)patients.Methods Liver tissues from 20 CHB patients as experimental group (CHB)and liver tissues of the surrounding hepatic benign adenomas from 20 cases as the control group (NC)were collected.The mRNA expressions of interleukin (IL)-17 and CD3γof liver tissues in each group were determined by real-time quantitative polymerase chain reaction (qPCR).The expressions of protein kinase B (Akt),phosphorylation-protein kinase B(p-Akt),p-S6 and IL-17 in each group were analyzed by Western blotting.With fluorescence labeled antibodies for immunohistochemical staining,the distributions of Th17 cells which co-expressed p-Akt and IL-17 protein of liver tissues in each group were observed and analyzed under fluorescent microscope. Hepatitis B virus (HBV )-related serological tests were measured.Data between two groups were compared by t test.Pearson correlation analysis was performed for correlation between groups.Results The expressions of IL-17 mRNA and CD3γmRNA in CHB group were both significantly increased than control group (22.530± 5 .657 vs 1 .025 ±0.246,t=16.99,P <0.01 ;12.791 ±3.433 vs 1 .033±0.305 ,t=15 .26,P <0.01).The ratio of IL-17 mRNA/CD3γ mRNA in CHB group (1 .782 ± 0.213 )was higher than that in control group (1 .014±0.144,t=13.49,P <0.01 ).The expression of IL-17 protein in CHB group (0.479 ±0.069 ) was significantly increased compared with control group (0.037 ±0.007,t =18.15 ,P <0.01 ).In CHB patients,the expression of IL-17 mRNA was positively correlated with the serum level of alanine aminotransferase (r=0.664,P <0.01),while there was no significant correlation between the expression of IL-17 mRNA and HBV viral load (r =0.053,P >0.05 ).The expressions of p-Akt and p-S6 in liver tissues of CHB patients were significantly increased compared with control group (0.355 ± 0.015 vs 0.145 ±0.009,t=30.93,P <0.01 ;0.380±0.043 vs 0.140±0.021 ,t=14.18,P <0.01 ),while there was no statistical difference of Akt expression between the two groups (t=2.10,P >0.05).The number of cells co-expressed IL-17 and p-Akt was significantly increased in CHB patients compared with control group (4.872 ± 0.513/high power field vs 1 .440 ± 0.301/high power field;t = 18.25 ,P < 0.01). Conclusions The activation of PI3K/Akt/mTOR signaling pathway and the expression of IL-17 indicate the activation of Th17 cells in the hepatic tissues of CHB,suggesting that PI3K/Akt/mTOR signaling pathway may play a role in the pathogenesis of CHB.