Published online Feb 15, 2003. doi: 10.3748/wjg.v9.i2.351
Revised: October 24, 2002
Accepted: November 4, 2002
Published online: February 15, 2003
AIM: To investigate how cholesterol (Ch) can affect the phenotype of bile duct fibroblasts of New Zealand rabbits.
METHODS: 16 rabbits were divided randomly into two groups: the control group and the experiment group. The rabbits in experiment group were fed with hypercholesterol diet for 8 wk. Bile duct was dissociated from rabbits and prepared for transmission electron microscopy. The purified bile duct fibroblasts were cultured and divided randomly into there groups: control group, Ch smiddle concentration group (0.6 g/L), Ch high concentration group (1.2 g/L). After incubated for 72 h, the fibroblasts were made into specimens for transmission electron microscopy. The expression of α-actin in bile duct fibroblasts was measured by means of laser scanning confocal microscopy.
RESULTS: With the transmission electron microscopy, the normal bile duct fibroblasts were shuttle-shaped, and there were abundant rough endoplasmic reticulums (RER), but few mitochondria or microfilaments in cytoplasm. This is the typical phenotype of fibroblasts. Bile duct fibroblasts of hypercholesterolemic rabbits were observed. by the transmission electron microscopy Rough endoplasmic reticulums were significantly reduced, with a lot of microfilament bundles or stress fibers appeared in cytoplasm, especially under plasma membrane. Dense bodies were scattered within these bundles. Macula densas and discontinuous sarcolemma were found under plasma membrane. It suggested that the bile duct fibroblasts of hypercholesterolemic rabbits presented the phenotype of smooth muscle cell. The cultured bile duct fibroblasts also had typical phenotype of fibroblasts. After stimulated by middle concentration cholesterol (0.6 g/L) for 72 h, there appeared lots of microfilaments in cytoplasm, but without dense body, macula densa and discontinuous sarcolemma. Observed with confocal microscopy, there were many regular bundles of microfilaments in fibroblasts treated with middle concentration ch (0.6 g/L) and the expression of α-actin was signifiantly increased. The average fluorescence value of middle concentration group was 1628 ± 189 (P < 0.01 versus control group). Microfilaments and the expression of α-actin were greatly decreased in fibroblastes of high concentration group (1.2 g/L). The average fluorescence value of high concentration group was 1427 ± 153 (P < 0.05 vs middle concentration group). There were a lower expression of α-actin and few microfilaments in bile duct fibroblasts of control group with an average fluorescence value of 1224 ± 138.
CONCLUSION: Cholesterol can make bile duct fibroblasts have the phenotypic characteristics of smooth muscle cell both in vitro and in vivo and this effect is more significant in vivo. The effect is probably associated with some other factors besides cholesterol.