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目的探讨泡球蚴组织蛋白在肝星状细胞活化中的作用。方法用不同浓度泡球蚴组织蛋白体外刺激人肝星状细胞,以PBS处理作为阴性对照,TGF-β1处理肝星状细胞作为阳性对照,分别采用qRT-PCR和Western blot检测肝星状细胞中COL1A1、α-SMA和TGF-β/Smad信号通路基因及蛋白表达水平。结果 60和600μg/ml泡球蚴蛋白处理肝星状细胞后α-SMA mRNA相对表达量分别为2.16±0.33和2.72±0.49,COL1A1mRNA相对表达量分别为1.73±0.12和5.39±0.14;TGF-β1处理肝星状细胞后α-SMA mRNA相对表达量为13.43±0.27,COL1A1mRNA相对表达量为18.89±2.59;PBS阴性对照组α-SMA基因相对表达量1.07±0.42,COL1A1基因相对表达量1.00±0.02。不同浓度泡球蚴蛋白组和TGF-β1处理组与PBS阴性对照组相比α-SMA及COL1A1mRNA相对表达量差异有统计学意义(P<0.05);肝星状细胞泡球蚴组织蛋白处理组与TGF-β1处理组α-SMA和COL1A1的蛋白水平较阴性对照组显著升高(P<0.05)。TGFβ-RII基因相对表达量在600μg/ml泡球蚴组织蛋白处理组为1.71±0.08,与PBS阴性对照组1.09±0.43相比差异具有统计学意义(P<0.05);Smad2基因相对表达量在TGF-β1处理组为1.68±0.29,与PBS阴性对照组1.07±0.42相比差异有统计学意义(P<0.05);Smad3基因相对表达量在600μg/ml泡球蚴组织蛋白处理组为2.41±0.84,TGF-β1处理组为1.61±0.15,与PBS阴性对照组1.00±0.09相比差异具有统计学意义(P<0.05)。结论泡球蚴组织蛋白中有类似TGF-β作用的能够诱发宿主肝星状细胞活化的细胞因子,该细胞因子可能参与泡球蚴感染所致宿主肝纤维化损伤。
Objective To explore the role of Echinococcus granulosus tissue protein in the activation of hepatic stellate cells. Methods Human hepatic stellate cells (HSCs) were stimulated in vitro with different concentrations of Echinococcus granulosus protein and treated with PBS as a negative control. TGF-β1-treated hepatic stellate cells served as positive controls and qRT-PCR and Western blot were used to detect hepatic stellate cells COL1A1, α-SMA and TGF-β / Smad signaling pathway genes and protein expression levels. Results The relative expression of α-SMA mRNA in hepatic stellate cells treated with 60 and 600 μg / ml alveolar protein was 2.16 ± 0.33 and 2.72 ± 0.49, respectively. The relative expression of COL1A1 mRNA was 1.73 ± 0.12 and 5.39 ± 0.14, respectively. TGF- The relative expression of α-SMA mRNA in hepatic stellate cells was 13.43 ± 0.27, and the relative expression of COL1A1 mRNA was 18.89 ± 2.59. The relative expression of α-SMA gene in PBS-negative control group was 1.07 ± 0.42 and the relative expression of COL1A1 gene was 1.00 ± 0.02 . The relative expression of α-SMA and COL1A1 mRNA in different concentrations of Echinocytes and TGF-β1-treated group was significantly lower than that in PBS-negative control group (P <0.05) The protein level of α-SMA and COL1A1 in TGF-β1 treatment group was significantly higher than that in negative control group (P <0.05). The relative expression of TGFβ-RII gene in 600μg / ml Echinococcus group was 1.71 ± 0.08, which was significantly different from 1.09 ± 0.43 in PBS-negative control group (P <0.05). The relative expression of Smad2 gene was The TGF-β1-treated group was 1.68 ± 0.29, which was significantly lower than that of PBS-negative control group (1.07 ± 0.42) (P <0.05). The relative expression level of Smad3 mRNA in the 600μg / ml Echinococcus group was 2.41 ± 0.84, and 1.61 ± 0.15 in TGF-β1-treated group, which was significantly different from 1.00 ± 0.09 in PBS-negative control group (P <0.05). Conclusion Cytokines of Echinococcus multilocularis may induce the activation of host hepatic stellate cells in a similar manner to that of TGF-β. These cytokines may be involved in the host liver fibrosis induced by Cysticercus cellulosae infection.