Effects of Kupffer cells stimulated by triglyceride and very low-density lipoprotein on proliferation of rat hepatic stellate cells.
OBJECTIVE
To study the effects of triglyceride, very low-density lipoprotein (VLDL), and Kupffer cell-conditioned medium (KCCM) derived from triglyceride and VLDL treatment on proliferation of rat hepatic stellate cells (HSC).


METHODS
HSC and Kupffer cells were isolated and cultured from liver of Wistar rats by in situ perfusion with proteinase and collagenase, and density gradient centrifugation with Nycodenz; HSC and Kupffer cells were identified by immunohistochemistry, endocytosis, and ultrastructure, etc. Kupffer cells were incubated with triglyceride (25 micrograms/ml) and VLDL (25 micrograms/ml) for 24 hours, KCCM were prepared, and MTT colorimetric assay was detected for HSC proliferation.


RESULTS
HSC proliferation was 0.1894 +/- 0.0316 (12.5 micrograms/ml), 0.1637 +/- 0.0243 (25 micrograms/ml), 0.1450 +/- 0.0264 (50 micrograms/ml), 0.1212 +/- 0.0275 (100 micrograms/ml), 0.1226 +/- 0.0138 (200 micrograms/ml) and 0.0990 +/- 0.0163 (400 micrograms/ml) in the presence of triglyceride and was 0.1583 +/- 0.0314 (6.25 micrograms/ml), 0.1642 +/- 0.0269 (12.5 micrograms/ml), 0.1834 +/- 0.0498 (25 micrograms/ml), 0.1964 +/- 0.0287 (50 micrograms/ml) and 0.2202 +/- 0.0284 (100 micrograms/ml) in presence of VLDL, respectively. Compared with the control, HSC proliferation at 400 micrograms/ml of triglyceride was lower (P < 0.01), but at 12.5 micrograms/ml of triglyceride and 25, 50, 100 micrograms/ml of VLDL higher (P < 0.05 or 0.01); HSC proliferation was 0.1569 +/- 0.0144, 0.1924 +/- 0.0113 and 0.1871 +/- 0.0116 in the presence of KCCM, KCCM + triglyceride and KCCM + VLDL, respectively. Compared with the control and KCCM, KCCM + triglyceride and KCCM + VLDL might promote HSC proliferation (P < 0.01); there was no statistical significance between KCCM + triglyceride and KCCM + VLDL (P > 0.05); KCCM was greater in HSC proliferation than the control, but there was no significant change (P > 0.05).


CONCLUSIONS
Triglyceride, VLDL, and KCCM stimulated by triglyceride and VLDL might promote HSC proliferation and be associated with fatty liver and hepatic fibrogenesis.
Lu
L
L
Shanghai Institute of Digestive Disease, Renji Hospital, Shanghai Second Medical University, Shanghai 200001, China.
Zeng
M
M
Li
J
J
Hua
J
J
Fan
J
J
Fan
Z
Z
Qiu
D
D
Xiao
S
S
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D017077:Culture Media, Conditioned; D022781:Hepatocytes / Q000166:cytology*; D007728:Kupffer Cells / Q000166:cytology*; D015227:Lipid Peroxidation; D008079:Lipoproteins, VLDL / Q000494:pharmacology*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D014280:Triglycerides / Q000494:pharmacology*
D017077:Culture Media, Conditioned; D008079:Lipoproteins, VLDL; D014280:Triglycerides