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Liver regenerated factor and its use

A technology for liver regeneration, reason, applied in the field of biological products and protein drugs, which can solve problems such as lack of stimulatory activity

Active Publication Date: 2007-01-17
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, many foreign laboratories have been working on the molecular cloning of such factors. In 1995, Hagiya et al. [3，4] A liver regeneration enhancing factor was isolated from rat regenerating liver tissue, and cloned and expressed. It was found that the recombinant liver regeneration enhancing factor could promote liver regeneration in partially hepatectomized rats, but lacked in vitro effects on primary cultured hepatocytes and Stimulatory activity of HCC cells

Method used

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  • Liver regenerated factor and its use
  • Liver regenerated factor and its use
  • Liver regenerated factor and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Production and preparation of HPPCn and its function of promoting liver cell proliferation and liver regeneration

[0024] Using DEAE-cellulose, Source15Q and other chromatography methods, SDS-PAGE segmented recovery, and MALDY-TOF and Q-TOF mass spectrometry techniques, a protein that can specifically promote hepatocyte proliferation and liver regeneration was purified and identified from newborn calf liver Factor, namely HPPCn. The human HPPCn cDNA sequence was obtained by screening the human fetal liver cDNA library. Add BamH I and Xhol I restriction sites at both ends of the gene sequence, construct it into the prokaryotic expression vector PBV220, and obtain the PBV220-HPPCn plasmid. It was transformed into Escherichia coli, and its expression was induced at 42°C. After ion exchange and gel filtration, the recombinant HPPCn with a purity of more than 95% is obtained, and its amino acid sequence is as follows: figure 1 shown.

[0025] 3 The proliferat...

Embodiment 2

[0029] Example 2 HPPCn to CCl 4 Protective effect of induced acute liver injury

[0030] Thirty Balbc mice in good condition were injected with 1ml / kg CCl 4 After that, they were randomly divided into 3 groups: Group I was given intravenous injection of 2.5 mg / kg HPPCn; Group II was given intravenous injection of 5 mg / kg HPPCn; Group III was given intravenous injection of normal saline. Inject every 12 hours, and observe the changes of ALT, AST and LDH in the blood after 48 hours. The results showed that the AST, ALT and LDH values ​​in serum of 48hr mice after injury were all significantly increased, but the AST, ALT and LDH values ​​of the mice in the HPPCn treatment group were significantly lower than those in the normal saline group (results shown in Figure 6), indicating that HPPCn to CCl 4 The induced acute liver injury has a protective effect.

Embodiment 3

[0031] Example 3 Inhibition of tumor cell growth in HPPCn cells

[0032] Construct HPPCn into the eukaryotic expression vector pEGFP-N1 plasmid, and transfect it into the human liver cancer cell line SMMC7721. After culturing for 36 hours, they were fixed with 4% paraformaldehyde and 70% ethanol, stained with PI, and FACS The changes in the cell cycle were detected, and the results showed that the transfected liver cancer cells showed obvious G0 / G1 phase arrest, and the proportion of cells in the G2 / M phase was significantly less than that of the control cells, suggesting that endogenous HPPCn had obvious growth inhibition on liver cancer cells (See Table 3).

[0033] cell

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Abstract

A liver textoblastic factor and its use are disclosed. It can be used to improve in vitro hepatic cell proliferation and in vivo liver neogenesis and tumor cell apoptosis, and inhibit tumor cell growth.

Description

technical field [0001] The present invention relates to the field of biological products and protein medicine, in particular to exogenous liver regeneration factor (hepatopoietin protein Cn, HPPCn) and its related protein (hepatopoietin protein Cn related protein, HPPCnr), these proteins can promote liver cell proliferation and liver regeneration , in cells, it can inhibit the growth of tumor cells and promote the apoptosis of tumor cells, which has potential clinical application value. Background technique [0002] The liver is an organ with a strong regenerative capacity in the body. The research on its regulatory mechanism has been conducted internationally for more than a hundred years. However, currently known growth factors related to liver regeneration such as hepatocyte growth factor (HGF), transforming growth factor-α TGF-α), etc., all lack liver specificity, and it is difficult to explain the mechanism of liver regeneration regulation with org...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475C12N15/12A61K38/18A61K31/7088A61K48/00A61P1/16A61P35/00
Inventor 吴祖泽石炳兴崔春萍杜邵君吴丹莉
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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