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目的研究泡型包虫病患者肝脏组织TGF-β1和Gadd45γ基因的表达及其在肝损伤中的作用,探讨其临床意义。方法采集23例AE患者肝脏手术标本,Trizol法提取RNA,反转录cDNA后采用实时荧光定量PCR检测TGF-β1、Gadd45γ、p53和p21基因的表达。体外培养肝细胞HL-7702,用重组TGF-β1(1ng/ml)细胞因子或匀浆虫体蛋白(1mg/ml)刺激30min、1h、24h和48h,分别收集细胞,采用实时荧光定量PCR检测TGF-β1、Gadd45γ、p53和p21基因的表达。结果 AE患者病灶旁肝脏组织TGF-β1、p53和p21mRNA的相对表达量分别为1.93±1.34、1.29±0.62和1.50±1.10,与远端肝组织1.00±0.00比较差异有统计学意义(P<0.05);Gadd45γmRNA为1.24±0.96,与远端组织比较差异无统计学意义(P>0.05)。Pearson法分析TGF-β1表达分别与Gadd45γ、p53和p21呈正相关(r值分别为0.5257、0.5287、0.4605,P<0.05)。用重组细胞因子TGF-β1(1ng/ml)或匀浆虫体蛋白(1mg/ml)刺激30min、1h、24h和48h,均能促进肝细胞HL-7702中TGF-β1、Gadd45γ、p53和p21基因的表达上调。结论在肝泡球蚴病晚期,炎症性细胞因子TGF-β1可能通过激活非经典型MAPK(Mitogen Activated Protein Kinase,JNK和p38)信号转导途径和细胞周期关键调控因子(Gadd45γ,p53,p21)调控肝脏细胞的生长抑制或凋亡,造成宿主肝脏的病理性损伤。
Objective To study the expression of TGF-β1 and Gadd45γ genes in liver tissues of patients with infiltrating hydatidosis and its role in liver injury, and to explore its clinical significance. Methods Twenty-three AE specimens from patients with AE were collected. Trizol method was used to extract RNA. Reverse transcriptase cDNA was used to detect the expression of TGF-β1, Gadd45γ, p53 and p21 genes by real-time fluorescence quantitative PCR. The hepatocytes HL-7702 were cultured in vitro, and the cells were harvested with recombinant TGF-β1 (1ng / ml) or homogenized protein (1mg / ml) for 30min, 1h, 24h and 48h, respectively. TGF-β1, Gadd45γ, p53 and p21 gene expression. Results The relative expression of TGF-β1, p53 and p21 mRNA in adjacent liver tissues of patients with AE were 1.93 ± 1.34, 1.29 ± 0.62 and 1.50 ± 1.10, respectively, which were significantly different from those of distal liver 1.00 ± 0.00 (P <0.05 ); Gadd45γ mRNA was 1.24 ± 0.96, which showed no significant difference compared with the distal tissue (P> 0.05). Pearson analysis showed that TGF-β1 expression was positively correlated with Gadd45γ, p53 and p21 (r values were 0.5257,0.5287,0.4605, P <0.05). TGF-β1, Gadd45γ, p53 and p21 in hepatocytes HL-7702 could be promoted by stimulation with recombinant cytokines TGF-β1 (1ng / ml) or homogenized worm protein (1mg / ml) for 30min, 1h, 24h and 48h Gene expression is up-regulated. CONCLUSION: TGF-β1, an inflammatory cytokine, may activate Gadd45γ, p53 and p21 signaling pathways by activating the signal transduction pathway of non-classical MAPK (JNK and p38) Regulation of liver cell growth inhibition or apoptosis, resulting in the pathological damage of the host liver.