Mechanisms of spontaneous resolution of rat liver fibrosis. Hepatic stellate cell apoptosis and reduced hepatic expression of metalloproteinase inhibitors.

Liver fibrosis results from the excessive secretion of matrix proteins by hepatic stellate cells (HSC), which proliferate during fibrotic liver injury. We have studied a model of spontaneous recovery from liver fibrosis to determine the biological mechanisms mediating resolution. Livers were harvested from rats at 0, 3, 7, and 28 d of spontaneous recovery from liver fibrosis induced by 4 wk of twice weekly intraperitoneal injections with CCl4. Hydroxyproline analysis and histology of liver sections indicated that the advanced septal fibrosis observed at time 0 (peak fibrosis) was remodeled over 28 d of recovery to levels close to control (untreated liver). alpha-Smooth muscle actin staining of liver sections demonstrated a 12-fold reduction in the number of activated HSC over the same time period with evidence of HSC apoptosis. Ribonuclease protection analysis of liver RNA extracted at each recovery time point demonstrated a rapid decrease in expression of the collagenase inhibitors TIMP-1 and TIMP-2, whereas collagenase mRNA expression remained at levels comparable to peak fibrosis. Collagenase activity in liver homogenates increased through recovery. We suggest that apoptosis of activated HSC may vitally contribute to resolution of fibrosis by acting as a mechanism for removing the cell population responsible for both producing fibrotic neomatrix and protecting this matrix from degradation via their production of TIMPs.

Laser Doppler flowmetry for determining changes in cutaneous blood flow following intradermal injection of histamine.

The use of laser Doppler flowmetry (LDF) in determining changes in cutaneous blood flow following intradermal injection of histamine has been investigated in this double-blind study. In eight subjects blood flow (LDF) and weal-and-flare area (planimetry) were measured at regular intervals for 1 hr following 50-microliter injections of different concentrations of histamine (6.5 X 10(-5)-6.5 X 10(-3)M) and saline. The mean maximum increase in LDF values over the flare was at least nine-fold greater than the baseline values for all three concentrations of histamine injected. When the LDF values observed at different sites were integrated to obtain the 'LDF response' it was possible to demonstrate concentration-related increases in blood flow and to differentiate clearly between the different concentrations of histamine and saline for up to 30 min after injection. During this period, the repeatability and the time course of the LDF response was comparable with that of the flare area. These studies suggest that the noninvasive technique of LDF is a sensitive and reproducible method for quantifying the changes in cutaneous blood flow that occurs for the first 30 min after intradermal injection of histamine.

Progelatinase A is produced and activated by rat hepatic stellate cells and promotes their proliferation.

Activated hepatic stellate cells (HSCs) are a potential source of gelatinase A, which accumulates in fibrotic livers. Progelatinase A activation requires its binding to a complex of membrane-type matrix metalloproteinase (MT-MMP) and tissue inhibitor of metalloproteinases (TIMP)-2. These studies examine gelatinase A, MT1-MMP, and TIMP-2 synthesis by HSCs during activation in vitro and the potential role of gelatinase A in promoting HSC proliferation. Gelatinase A, MT1-MMP, and TIMP-2 messenger RNA (mRNA) were all upregulated in HSCs activated on plastic over 5 to 14 days. Gelatinase A expression was maximal at 7 days of culture, coinciding with the peak of HSC proliferation and the onset of procollagen I and alpha-smooth muscle actin (alpha-SMA) mRNA expression. Active forms of gelatinase A of 62 kd and 66 kd were secreted by activated HSCs and reached a maximum of 12.1% of total enzyme in 14-day culture supernatants. Treatment of HSCs with concanavalin A (con A) induced activation of MT1-MMP and enhanced secretion of activated gelatinase A, which reached a maximum of 44.4% of the total enzyme secreted into culture supernatants using 30 microgram/mL con A. [(14)C]-gelatin degradation assays confirmed the presence of gelatinolytic activity in activated HSC supernatants, which reached a maximum level at 7 days of culture. Antisense oligonucleotide inhibition of endogenous progelatinase A production, or the MMP inhibitor 1,10-phenanthroline inhibited (3)H-thymidine incorporation into HSC DNA by greater than 50%. We conclude that HSCs produce progelatinase A during activation in vitro and activate this enzyme coincident with MT1-MMP and TIMP-2 synthesis. Gelatinase A activity is required for maximal proliferation of HSCs in vitro suggesting this metalloproteinase is an autocrine proliferation factor for HSCs.

High prevalence of coeliac disease in a population-based study from Western Australia: a case for screening?

OBJECTIVES: To determine the prevalence of coeliac disease in an Australian rural community. DESIGN: Retrospective analysis of stored serum samples from 3,011 random subjects from the Busselton Health Study. IgA antiendomysial antibodies (AEA) were detected by indirect immunofluorescence, and subjects testing positive were contacted and offered small-bowel biopsy. MAIN OUTCOME MEASURES: Prevalence of AEA positivity and biopsy-proven coeliac disease in the community with reference to the proportion of symptomatic to asymptomatic patients. RESULTS: 10 of 3,011 subjects were AEA positive. One subject had died, one subject could not be traced and one refused small-bowel biopsy. All subjects with detectable AEA who consented to biopsy had pathological changes consistent with coeliac disease. The prevalence of newly diagnosed biopsyproven coeliac disease is 7 in 3,011 (1 in 430). Two further subjects had a diagnosis of coeliac disease before this study. When all AEA-positive patients and those previously diagnosed are included, the prevalence is 12/3,011 (1 in 251). There was a significant clustering of cases in the 30-50-years age range, with 10/12 (83%; 95% CI, 52%-98%) aged between 30 and 50 years, compared with 1,092/3,011 (36%; 95% CI, 35%-38%) of the total population (P<0.03). Of the eight AEA-positive subjects who could be contacted, four had symptoms consistent with coeliac disease and four were asymptomatic. Three subjects were iron-deficient, four subjects had first-degree relatives with coeliac disease and one subject had type 1 diabetes mellitus. CONCLUSIONS: The prevalence of coeliac disease is high in a rural Australian community. Most patients are undiagnosed, and asymptomatic.