β-Actin: Not a Suitable Internal Control of Hepatic Fibrosis Caused by Schistosoma japonicum

Abstract

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. β-actin, a traditional housekeeping gene, is widely used as an internal control to normalize gene and protein expression. However, β-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 β-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 β-Tubulin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) remained stable. The dynamic change of β-actin was similar with the profibrogenic factors, including α-SMA, Collagen I, and Collagen III. We employed immunofluorescence staining and further showed that the expression level of β-actin was positively correlated with α-SMA. What is more, there was a positive correlation between the level of β-actin mRNA and the content of hydroxyproline in liver. This study provides evidences that β-actin is variable and unsatisfied for application as an internal control in hepatic fibrosis induced by S. japonicum infection.

Hepatic Progenitor Cells Promote the Repair of Schistosomiasis Liver Injury Through Inhibiting IL-33 Secretion 

“…The expression levels of HGF mRNA, IL-6 mRNA, α-SMA mRNA, Collagen-I mRNA, IL-33 mRNA and GAPDH mRNA were detected using SYBR Premix Ex TaqTM (TaKaRa, Osaka, Japan) and the speci c primers pairs were listed in Table 1. The reaction procedure was followed as described previously [11] and performed with a CFX 96 touch instrument (Bio-Rad, CA, United States). The GAPDH mRNA level was used for normalization.…”

Necroptosis and Caspase-2-Mediated Apoptosis of Astrocytes and Neurons, but Not Microglia, of Rat Hippocampus and Parenchyma Caused by Angiostrongylus cantonensis Infection

“…Consecutive formalin-fixed paraffin-embedded sections of the brains of the animals were subjected to immunostaining as previously described (Zhang et al, 2019). Briefly, PBS (phosphate buffered saline) containing 1% bovine serum albumin (BSA) was used to block the sections for 1 h, followed by incubation with an anti-RIP3 (Santa Cruz, CA, United States) primary antibody overnight at 4 • C. After 3 washes with PBS and 0.1% Tween-20 for 5 min, the sections were stained for 1 h with rhodamine-labeled goat anti-mouse lgG (Abways Technology, Shanghai, China), and the cell nuclei were stained with 4'6-diamidino-2-phenylindole for 5 min.…”

The Differential and Dynamic Progression of Hepatic Inflammation and Immune Responses During Liver Fibrosis Induced by Schistosoma japonicum or Carbon Tetrachloride in Mice

“…Dynamic progressions of pathological changes, the cytokines and lymphocytes subsets in mice infected by S. japonicum have been reported in several reports with various results (15)(16)(17)(18), while those in CCl 4 induced liver fibrosis were rarely reported or detected at different time point (19,20). It was difficulty to make comparisons between the two models from the previous literatures.…”