β-Actin: Not a Suitable Internal Control of Hepatic Fibrosis Caused by Schistosoma japonicum
β-Actin: Not a Suitable Internal Control of Hepatic Fibrosis Caused by Schistosoma japonicum
复制标题
β-肌动蛋白：不是日本血吸虫引起的肝纤维化的合适内部对照
DOI：
10.3389/fmicb.2019.00066
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发表时间：
2019-01-31
影响因子：
5.2
通讯作者：
Wu, Zhongdao
中科院分区：
文献类型：
--
作者：
Zhang, Beibei;Wu, Xiaoying;Wu, Zhongdao
日本血吸虫病是一个导致人类发病和死亡的重大健康问题。它的特征是由虫卵在肝脏沉积引起的肝脏肉芽肿反应和纤维化。β - 肌动蛋白是一种传统的管家基因，被广泛用作内参来校正基因和蛋白质的表达。然而，在使用药物治疗或某些生理和病理条件下，β - 肌动蛋白的表达可能会波动。在本研究中，我们发现日本血吸虫感染6周后，随着肝纤维化的形成，β - 肌动蛋白的mRNA和蛋白质表达均显著增加，并且在肝纤维化进展过程中保持高水平，而β - 微管蛋白和甘油醛 - 3 - 磷酸脱氢酶（GAPDH）的水平保持稳定。β - 肌动蛋白的动态变化与促纤维化因子（包括α - 平滑肌肌动蛋白、I型胶原蛋白和III型胶原蛋白）相似。我们通过免疫荧光染色进一步表明，β - 肌动蛋白的表达水平与α - 平滑肌肌动蛋白呈正相关。此外，β - 肌动蛋白mRNA水平与肝脏中羟脯氨酸含量呈正相关。本研究提供的证据表明，β - 肌动蛋白是可变的，不适合作为日本血吸虫感染诱导的肝纤维化研究中的内参。
Schistosomiasis japonica is a significant health problem that leads to morbidity and mortality of humans. It is characterized by hepatic granulomatous response and fibrosis caused by eggs deposition in the liver. beta-actin, a traditional housekeeping gene, is widely used as an internal control to normalize gene and protein expression. However, beta-actin expression can fluctuate upon the treatment with pharmacological agents or under some physiological and pathological conditions. In this study, we found that the expressions of both beta-actin mRNA and protein increased significantly with hepatic fibrosis formation after 6 weeks infection with Schistosoma japonicum and kept high level during the progression of hepatic fibrosis, while the levels of beta-Tubulin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) remained stable. The dynamic change of beta-actin was similar with the profibrogenic factors, including alpha-SMA, Collagen I, and Collagen III. We employed immunofluorescence staining and further showed that the expression level of beta-actin was positively correlated with alpha-SMA. What is more, there was a positive correlation between the level of beta-actin mRNA and the content of hydroxyproline in liver. This study provides evidences that beta-actin is variable and unsatisfied for application as an internal control in hepatic fibrosis induced by S. japonicum infection.