Hepatopoietin Cn (HPPCn) Generates Protective Effects on Acute Liver Injury.

Na Li,Mei-Hua Qu,Chun-Xia Sun,Jian Li,Dan-Dan Li,Feng-Jiao Liu,Chun-Ping Cui,Da-Jin Zhang

doi:10.3389/fphar.2019.00646

Abstract

Objective: To observe the protective role of hapatopoietin Cn (HPPcn) on acute liver injury. Methods: Six hours after 10 mmol/L CCl4, 150 mmol/L ethanol, or 0.6 mmol/L H2O2 treatment, SMMC7721 human hepatoma cells were incubated with 10, 100, or 200 ng/ml recombinant human HPPCn protein (rhHPPCn) for an additional 24 h. The cell survival rate was analyzed using the CCK-8 assay. The CCl4-induced apoptosis of SMMC7721 cells was detected by flow cytometry. Then, the levels of glutamic oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), malondialdehyde (MDA), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD) in SMMC7721 cell lysates and cell culture supernatant were detected. SMMC7721 cells were treated with different concentrations of rhHPPCn (0, 10, and 100 ng/ml). The cell proliferation indexes (BrdU incorporation and PCNA expression) were detected by immunohistochemistry (IHC). An acute liver injury mouse model was established by a one-time intraperitoneal injection of 20% CCl4 at a volume of 5 ml/kg body weight. One hour after CCl4 injection, 1.25 or 2.5 mg rhHPPCn/12 h/kg body weight was injected via the tail vein. The serum levels of GOT and GPT were detected at different time points. Pathological changes in the liver were evaluated. PCNA expression levels were observed by IHC. Results: rhHPPCn increased the survival rate of SMMC7721 cells and inhibited chemical toxicity-induced cell apoptosis. The levels of GOT, GPT, MDA, and LDH in the cell supernatant were significantly reduced, while GSH-PX and SOD were significantly increased after rhHPPCn treatment in the CCl4-treated SMMC7721 cells. BrdU incorporation and PCNA expression increased in a concentration-dependent manner, indicating that rhHPPCn promotes cell proliferation. The results showed that rhHPPCn significantly reduced the serum levels of GOT and GPT in CCl4-induced acute liver injury mice. rhHPPCn alleviated the tissue damage and increased PCNA expression, indicating the promotion of proliferation after acute injury. Conclusion: rhHPPCn protects hepatocytes from chemical toxins by promoting proliferation and inhibiting apoptosis in vivo and in vitro. Our study provides new insights for the clinical treatment of acute liver injury.

Highlights

Hepatopoietin Cn (HPPCn) is a growth factor that initially isolated from the hepatic stimulator substance (HSS) of newborn calf, which promotes hepatocyte proliferation
The results showed that recombinant human HPPCn protein (rhHPPCn) significantly increased BrdU
The results suggested that rhHPPCn could promote DNA synthesis in SMMC7721 cells after CCl4-induced acute injury

Summary

Introduction

Hepatopoietin Cn (HPPCn) is a growth factor that initially isolated from the hepatic stimulator substance (HSS) of newborn calf, which promotes hepatocyte proliferation. ANP32 family members are involved in various biological processes, such as cell proliferation, differentiation, and apoptosis, and are related to tumorigenesis and drug resistance (Huyton and Wolberger, 2007). It was reported that recombinant human HPPCn protein (rhHPPCn) can stimulate hepatocyte proliferation in vitro and in vivo by activating signaling pathways that include sphingosine kinase-1 and extra cellular signal-regulated kinase (Chang et al, 2010; Liu et al, 2013). Most of the factors target a variety of cells and organs, lacking specificity for the liver regeneration process. RhHPPCn is a well-defined hepatocyte growth factor with specific hepaticstimulating activities in partially hepatectomized (PH) mice (Cui et al, 2008; Liu et al, 2013). HPPCn expression is significantly increased in the liver of PH mice and rhHPPCn can stimulate DNA replication and Erk phosphorylation in hepatocytes of PH mice (Cui et al, 2008)

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Conclusion