A.G. Lemmens

Sterol balance and cholesterol absorption in inbred strains of rabbits hypo- or hyperresponsive to dietary cholesterol

In 2 inbred strains of rabbits with high or low response of plasma cholesterol to dietary cholesterol, excretion of steroids in the feces and efficiency of cholesterol absorption were determined. Rates of whole-body cholesterol synthesis, measured as fecal excretion of bile acids and neutral steroids minus cholesterol intake, were similar in hypo- and hyperresponders fed a low-cholesterol (8 mumol/100 g) diet. Transfer of the rabbits to a high-cholesterol (182 mumol/100 g) diet caused an increase in fecal bile acid excretion in hypo- but not in hyperresponders. Dietary cholesterol did not affect neutral steroid excretion in either rabbit strain. Hyperresponders tended to accumulate more cholesterol in their body than did hyporesponders. After the rabbits were switched back from the high- to the low-cholesterol diet, rates of whole-body cholesterol synthesis were significantly higher in the hypo- than in the hyperresponders. With the use of the simultaneous oral administration of [3H]cholesterol and beta-[14C]sitosterol, hyperresponders were found to absorb significantly higher percentages of cholesterol than hyporesponders. It is concluded that the differences in stimulation of bile acid excretion after cholesterol feeding and the efficiency of cholesterol absorption are important determinants of the phenomenon of hypo- and hyperresponsiveness in the 2 inbred rabbit strains.

Cholesterol metabolism and esterases in four strains of rats with differential cholesterolemic responses to a high-cholesterol, high-cholate diet.

The increase in serum cholesterol after feeding a diet containing 2% (w/w) of cholesterol and 0.5% of cholate for 13 days was 200 and 800% in two hypo- and two hyper-responsive inbred strains of rats, respectively. While remaining on the high-cholesterol, high-cholate diet for longer periods, the level of serum cholesterol dropped in the hyper-responsive strains, and after 8 weeks on the diet one hyper-responsive strain had similar serum cholesterol concentrations as the two hypo-responsive strains. The feeding of a semipurified diet, containing 1% (w/w) of cholesterol and 20% of fat, did not discriminate between the two hypo- and hyper-responsive strains with respect to the response of serum cholesterol. The activities in plasma of the indicators for liver function, aspartate amino transferase and alkaline phosphatase, were significantly increased in all strains after feeding the high-cholesterol, high-cholate diet. Only alkaline phosphatase was increased by the semipurified diet. Evidence is presented that in the four inbred strains of rats the differential cholesterolemic response to the high-cholesterol, high-cholate diet is not related to the baseline serum lipoprotein profile, liver cholesterol accumulation, fecal bile acid excretion, and the total activities and patterns of esterases in serum, liver and small intestine.

Co-variance of chemically and histologically analysed severity of dystrophic cardiac calcification in mice

Dystrophic cardiac calcification (DCC) is a post-mortem finding in mice of various strains frequently used in biomedical research. The major aim of this study was to see whether DCC severity can be assessed by chemical analysis of calcium in the heart. Histological examination was used as the method of reference. Hearts of mice of four strains (BALB/c, C3H, C57BL/6 and DBA/2) were halved and the two resulting parts were subsequently subjected to histology or chemical analysis. Within hearts, the halves generally yielded similar results. The DCC scores and calcium contents were directly correlated within hearts. Thus, calcium analysis could serve as an alternative to histological examination in the assessment of DCC severity in mice. DBA/2 and C3H mice were found to be affected by DCC. Plasma magnesium concentrations were lower in these strains than in the DCC-free C57BL/6 and BALB/c strains. The tongue, lungs and diaphragm were also found to be calcified in DCC positive animals. Possibly, DCC is just one component of a generalized soft tissue calcification.

Esterases in inbred strains of mice with differential cholesterolemic responses to a high-cholesterol diet.

Specific esterase isoenzyme patterns in plasma may be associated with responsiveness of serum cholesterol to dietary cholesterol. In rabbits and rats the presence and absence of a high-mobility, anodal esterase band on electrophoresis have been shown to be associated with hypo- and hyperresponsiveness, respectively. We fed for 28 days male mice of 7 inbred strains either a low-cholesterol, commercial diet or a diet containing 2% (w/w) cholesterol, 0.5% cholic acid and 5% olive oil. Feeding the high-cholesterol diet revealed marked inter-strain differences in the responses of plasma and liver cholesterol; the increases ranged from 21 to 129% and from 10 to 80-fold, respectively. There was no association between esterase isoenzyme patterns in plasma and the sensitivity to the high-cholesterol diet. The mean baseline plasma total esterase activity tended to be positively associated with the absolute response of plasma cholesterol to the high-cholesterol diet (r = 0.56; n = 7), but the positive relationship between the baseline concentration of the ES-1 component in plasma and the cholesterolemic response was stronger (r = 0.84; n = 7; P less than 0.05). The high-cholesterol diet caused a significant increase in plasma total esterase activities in 6 out of the 7 strains. Evidence is presented that the increase in plasma total esterase activity, which was associated with an increase in the activity and concentration of the so-called ES-2 isoenzyme, is the result of an enhanced release of esterases from the intestine, rather than from the liver. A significant, positive correlation was found between the baseline intestinal esterase activity and the cholesterolemic response after cholesterol feeding (r = 0.83; n = 7; P less than 0.05).

The hypercholesterolemic effect of dietary coconut fatversus corn oil in hypo- or hyperresponsive rabbits is not exerted through influencing cholesterol absorption

In two inbred strains of rabbits with high or low response of plasma cholesterol to dietary saturatedversus polyunsaturated fatty acids, the efficiency of intestinal cholesterol absorption was measured. The feeding of a cholesterol-free purified diet containing saturated fatty acids in the form of coconut fat, when compared with a diet containing corn oil as polyunsaturated fatty acids, did not influence the efficiency of cholesterol absorption in the two rabbit strains. Irrespective of the dietary fat source, the hyperresponsive rabbits absorbed cholesterol more efficiently. It is concluded that the hypercholesterolemic effect of dietary coconut fatversus corn oil is not exerted by influencing cholesterol absorption.

Phosphorus-induced nephrocalcinosis in female rats: a study on regression and clinical abnormalities.

The question addressed was whether pre-established phosphorus (P)-induced nephrocalcinosis would regress after dietary P restriction. Female rats were fed purified diets containing either 0·2% (w/w) P(low P) or 0·6% P (high P). After 29 days, the high-P diet had caused massive nephrocalcinosis as demonstrated chemically (by the analysis of calcium in kidney) and histologically Jby inspection of kidney sections stained for calcium phosphate deposits). Switching rats from the high P to the low P diet did not result in a decrease in the degree of nephrocalcinosis within 91 days. Thus, P-induced nephrocalcinosis may not regress upon subsequent P restriction. Rats that had been fed either the 0·2 or 0·6% P diet for 56 days were examined clinically with respect to 14 selected variables. None of the variables discriminated between rats with or without nephrocalcinosis. This might imply that P-induced nephrocalcinosis in female rats does not cause significant discomfort.

Rabbit Models Hypo- or Hyperresponsive to Changes in Diet

The feeding of a high-cholesterol diet to random-bred rabbits generally elicits marked differences in the response of plasma cholesterol between individuals. Individuals showing only small changes in the concentrations of plasma cholesterol (hypores-ponders) can be discriminated from rabbits showing high degrees of hypercholesterolemia (hyperresponders). The mechanisms underlying hypo- and hyperrespon-siveness have not yet been unravelled, but the availability of inbred strains of rabbits with defined, but different cholesterolemic responses to changes in diet may be of great importance in this respect. Table 1 shows that certain strains of rabbits are relatively sensitive or insensitive to cholesterol loading. In this communication we present the results of further studies with the strains displaying the most extreme (AX/J) and lowest response (IIIVO/J).