Constitutive expression and regulation of rat complement factor H in primary cultures of hepatocytes, Kupffer cells, and two hepatoma cell lines

Lab Invest. 2002 Feb;82(2):183-92. doi: 10.1038/labinvest.3780410.

Abstract

The 155-kd soluble complement regulator factor H (FH), which consists of 20 short consensus repeats, increases the affinity of complement factor I (FI) for C3b by about 15 times. In addition to its cofactor activity, it prevents factor B from binding to C3b and promotes the dissociation of the C3bBb complex. The primary site of synthesis of FH, as well as of FI, is the liver, but the cell types responsible for the hepatic synthesis of both factors have not yet been clearly identified. In contrast to FI-mRNA, which was detectable only in hepatocytes (HC), FH-specific mRNA was identified in both HC and Kupffer cells (KC). As calculated for equal amounts of mRNA isolated from both cell types, FH-specific mRNA was found to be nearly 10-fold higher in KC than in HC, leading to the conclusion that KC are an abundant source of FH. Of the investigated proinflammatory cytokines IL-6, TNF-alpha, IL-1beta, and IFN-gamma, only IFN-gamma up-regulated FH-specific mRNA up to 6-fold in both primary HC and KC. This was also demonstrable on the protein level. However, FH-specific mRNA was not inducible in the rat hepatoma cell line H4IIE, which did not express FH-specific mRNA and could not be up-regulated in FAO cells that constitutively expressed FH-specific mRNA. This demonstrates that transformed cell lines do not reflect FH regulation in isolated primary HC. In addition to IFN-gamma, the endotoxin lipopolysaccharide (LPS) up-regulated FH-specific mRNA nearly 10-fold in KC after stimulation at concentrations of 10 or 1 ng/ml. In contrast, concentrations of up to 2 microg LPS/ml did not show any effect on HC. Our data suggest that LPS does not regulate the expression of FH in HC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Cells, Cultured
  • Complement Factor H / genetics
  • Complement Factor H / metabolism*
  • DNA Primers
  • Dose-Response Relationship, Drug
  • Hepatocytes / metabolism*
  • Interferon-gamma / pharmacology
  • Interleukin-1 / pharmacology
  • Interleukin-6 / pharmacology
  • Kupffer Cells / metabolism*
  • Lipopolysaccharides / pharmacology
  • Liver Neoplasms, Experimental / metabolism*
  • Male
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • Rats
  • Rats, Wistar
  • Tumor Cells, Cultured
  • Up-Regulation / drug effects

Substances

  • DNA Primers
  • Interleukin-1
  • Interleukin-6
  • Lipopolysaccharides
  • RNA, Messenger
  • Complement Factor H
  • Interferon-gamma