Immunotherapy for nonalcoholic steatohepatitis using the multiple cytokine production modulator Y-40138.
OBJECTIVE
To investigate the possible use of the multiple cytokine production modulator, Y-40138, as a novel immunotherapy in the rat nonalcoholic steatohepatitis (NASH) model.


METHODS
We allocated 6-wk-old male F344 rats to choline-supplemented, L-amino acid-defined (CSAA) diet (control group), CSAA diet + Y-40138 (control + Y-40138 group), choline-deficient, L-amino acid-defined (CDAA) diet (NASH group), or CDAA diet + Y-40138 (NASH + Y-40138 group). In each group, we measured the plasma alanine aminotransferase (ALT) levels, and the plasma and liver levels of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and interleukin-10 (IL-10). Tissue specimens of phosphate buffered saline-perfused liver were subjected to hematoxylin and eosin staining, Azan staining, Sirius red staining, and immunohistochemical staining (for Kupffer cells and TNF-alpha). We then extracted Kupffer cells from the collagenase-perfused livers using the Percoll gradient centrifugation method, and measured the TNF-alpha levels in the supernatant (in vitro TNF-alpha production by Kupffer cells) using an enzyme-linked immunosorbent assay kit.


RESULTS
In comparison to the NASH group, serum ALT elevation was mild, production of serum and liver TNF-alpha and IFN-gamma was inhibited, and IL-10 production was increased in the NASH + Y-40138 group. Amelioration of liver histology was also noted in the NASH + Y-40138 group. Kupffer cell immunohistochemical staining revealed no differences between groups, whereas TNF-alpha immunohistochemical staining showed fewer stained cells in the NASH + Y-40138 group than in the NASH group. The TNF-alpha levels in the in-vitro Kupffer cell culture supernatant were lower in the NASH + Y-40138 group than in the NASH group.


CONCLUSIONS
Administration of Y-40138 to NASH model rats reduced hepatic inflammation and suppressed fibrosis. These results indicate that the multiple cytokine production modulator, Y-40138, is promising as a novel treatment for NASH.
Tsujimoto
Tatsuhiro
T
Third Department of Internal Medicine, Nara Medical University, Kashihara, Nara, Japan. tat-tyan@xa2.so-net.ne.jp
Kawaratani
Hideto
H
Kitazawa
Toshiyuki
T
Yoshiji
Hitoshi
H
Fujimoto
Masao
M
Uemura
Masahito
M
Fukui
Hiroshi
H
D000081:Acetamides / Q000494:pharmacology*; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D017364:Collagenases / Q000737:chemistry; D016207:Cytokines / Q000037:antagonists & inhibitors* / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005234:Fatty Liver / Q000628:therapy*; D007150:Immunohistochemistry / Q000379:methods; D007167:Immunotherapy / Q000379:methods*; D007371:Interferon-gamma / Q000097:blood; D016753:Interleukin-10 / Q000097:blood; D007728:Kupffer Cells / Q000187:drug effects; D008297:Male; D010879:Piperazines / Q000494:pharmacology*; D051381:Rats; D011916:Rats, Inbred F344; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
D000081:Acetamides; D016207:Cytokines; D010879:Piperazines; D014409:Tumor Necrosis Factor-alpha; C406663:Y 39041; D016753:Interleukin-10; D007371:Interferon-gamma; D000410:Alanine Transaminase; D017364:Collagenases