The amount of fructose in 2000 kcal diets was approximately 80 g (high fructose diet) and 10 g (low fructose diet)... Throughout the study, fasting or postprandial
plasma TG of women was not affected by consumption of either diet.
However, over the course of the study, fasting
plasma TG decreased in both groups (with respect to baseline).
Lowering of
plasma TG levels after injection of [3H] triolein - labeled Liposyn particles was associated with a greater uptake of [3H] triolein in the liver, but not in organs such as the heart or skeletal muscle (Figure 8F), as would be expected if LPL activity were enhanced.
(E)
Plasma TG levels of fasted mice before and after injection of 100 μl 20 % Liposyn II particles at the indicated time points.
Together, these findings suggest that the major mechanism by which ApoC - III affects
plasma TG levels functions independently of LPL.
As reported by Noh et al. (43), tamoxifen transiently increases
plasma TG levels, which return to normal 2 weeks after the last administration.
Again, ApoC - III ASO treatment had no effect on
plasma TG levels in mice lacking both Lrp1 and Ldlr (Figure 5, A and C), as observed with animals fed a standard chow diet (Figure 1) and under postprandial conditions (Figure 3).
ApoC - III ASO — mediated
plasma TG reduction in mice on a chow diet depends on hepatic LDLR and LRP1.
These findings suggest that an ASO - mediated reduction of
plasma TG levels depends on LDLR and LRP1.
We also present evidence that TRLs with reduced ApoC - III content do not affect lipolysis and that a reduction in LPL expression and activity does not suppress the impact of ASO inhibition on lowering
plasma TG levels.
However, an increase in
plasma TG levels also occurs in ApoC - III — transgenic mice that are deficient in ApoE (20, 35).
Administration of the ApoC - III ASO improves
plasma TG independently of changes in LPL activity or VLDL secretion.
ApoC - III ASO — mediated
plasma TG reductions in mice on a high - fat diet depends on hepatic LDLR and LRP1.
However, a recent clinical study showed that lowering plasma ApoC - III levels with volanesorsen, a generation 2.0 + ApoC - III — specific antisense oligonucleotide (ASO), dramatically lowered elevated
plasma TG levels in patients who have genetic defects in LPL (familial chylomicronemia syndrome [FCS]-RRB-(16).
In chow - fed animals, ApoC - III ASO reduced fasting
plasma TG levels by 30 % to 45 % in animals defective in hepatic Ndst1 and Ldlr and in animals with combined deletions of Ndst1 and Lrp1 or Ldlr (Figure 1, D and E).
Nevertheless, the reduction in
plasma TG and cholesterol levels induced by ApoC - III ASO did not enhance hepatic steatosis (Figure 5, E and F) or induce weight gain or loss in the mice (Supplemental Figure 4).
However, the ASO had no effect on
plasma TG levels in mice lacking both Lrp1 and Ldlr (Ldlr — / — Lrp1fl / fl Alb - Cre +)(Figure 1, D and E).
Elevated
plasma TG levels represent a risk factor for CVD in humans, and an adjusted analysis of patients in the Framingham Heart Study suggests that each decrease of 1 mg / dl in plasma ApoC - III levels is associated with a 4 % decrease in CVD risk (11), further supporting the potential therapeutic benefit of suppressing plasma ApoC - III levels in humans to reduce CVD.
Plasma TG levels were determined 2 weeks after the last tamoxifen injection.
(D) Fasting
plasma TG levels in mutant mice treated for 4 weeks with control or ApoC - III ASO (n = 5 — 10 / group).
The high - fat diet raised
plasma TG levels in Ldlr — / — , Ldlr — / — Ndst1fl / fl Alb - Cre +, and Ldlr — / — Lrp1fl / fl Alb - Cre + mice, reaching values of 515 to 1,045 mg / dl (Figure 5A).
Blood samples were collected at the indicated time points and processed to measure
plasma TG accumulation and VLDL production rates (n = 3 / group).
Similarly, inactivating mutations affecting the expression of APOC3 in humans led to reduced
plasma TG levels (8) and protection against CVD (9 — 11).
The iLpldf mice presented with elevated
plasma TG and cholesterol levels (Supplemental Figure 5, C and D, respectively), along with a dramatic increase in VLDL TGs and a reduction in HDL cholesterol levels (Supplemental Figure 5, E and F), as observed in patients with LPL deficiency (16).
A comparable reduction in
plasma TG levels occurred after heparin injection of mice treated with ApoC - III ASO (Figure 7A).
Graham et al. showed that lowering ApoC - III using antisense oligonucleotides reduces
plasma TGs in rodents, nonhuman primates, and humans (15).
To examine whether hepatic clearance via these receptors contributed to an ApoC - III ASO — mediated reduction of
plasma TGs, we administered the ApoC - III ASO for 4 weeks to mice lacking LDLR (Ldlr — / ---RRB-, HSPGs (Ndst1fl / fl Alb - Cre +), or hepatic LRP1 (Lrp1fl / fl Alb - Cre +), and to mice lacking various pairs of these receptors (depicted in Figure 1B).
Perhaps blocking the rapid clearance of TRL remnants through LDLR and LRP1 has an adaptive role in shunting fatty acids liberated from
plasma TGs to the heart and skeletal muscle when food is less plentiful.
We used a previously described tamoxifen - inducible, Lpl - deficient mouse model (Lplfl / fl Actb - MerCreMer +, referred to herein as iLpldf mice) to further assess the in vivo contribution of LPL to the reduction of
plasma TGs by ApoC - III ASO (22).
(A and B)
Plasma TGs were measured before and after heparin injection in WT, Ldlr — / — Ndst1fl / fl Alb - Cre +, and Ldlr — / — Lrp1fl / fl Alb - Cre + mice treated for 4 weeks with control or ApoC - III ASO (A) after an overnight fast or (B) 3 hours after a fat challenge.
Repetitive treatment with the ApoC - III ASO progressively decreased
plasma TGs (Supplemental Figure 6B) and increased TG clearance after oral corn oil gavage (Supplemental Figure 6C) or after injection of [3H] triolein - labeled Liposyn particles (Figure 8E).
Under these conditions, administration of the ApoC - III ASO reduced hepatic Apoc3 mRNA expression in the various mutants by 74 % to 99 % (Figure 5B) and reduced
plasma TGs by 35 % to 57 % (Figure 5, A and C).
Treatment with ApoC - III ASO reduced postprandial
plasma TGs in all of the mutants except in Ldlr — / — Lrp1fl / fl Alb - Cre + mice (Figure 4, A and B).
Treatment of iLpldf mice with ApoC - III ASO decreased Apoc3 mRNA levels (Figure 8A) and protein expression (Supplemental Figure 6A) and reduced
plasma TGs (Figure 8B) and TGs in CR / VLDL particles (Figure 8C).
ApoC - III ASO treatment lowered
plasma TGs in mice lacking lipoprotein lipase (LPL), hepatic heparan sulfate proteoglycan (HSPG) receptors, LDLR, or LRP1 and in animals with combined deletion of the genes encoding HSPG receptors and LDLRs or LRP1.
Antisense oligonucleotides (ASOs) for ApoC - III reduce
plasma TGs in primates and mice, but the underlying mechanism of action remains controversial.
We show that an ApoC - III ASO — mediated reduction of
plasma TGs is mediated predominantly through inhibition of hepatic TRL clearance via the LDL / LRP1 axis.
Simultaneous treatment with Apoc3 ASO significantly reduced hepatic Apoc3 mRNA in all models (Supplemental Figure 1C) and reduced
plasma TGs by 20 % to 35 % in mice treated with saline, LDLR ASO, or LRP1 ASO (Supplemental Figure 1, D and E).
These results are consistent with the recent observation that targeting of ApoC - III effectively reduces
plasma TGs in LPL - deficient patients (16).
Not exact matches
Of note, the
plasma glucose values obtained for our untreated
Tg - hIAPP animals were a bit lower than those reported by Janson et al. (1996).
Total cholesterol and
TG levels in
plasma were determined using kits from Genzyme or Thermo Fisher Scientific.
This residual risk has shifted attention to elevated
plasma triglyceride (
TG) levels, which constitute an independent risk factor for coronary artery disease.
To measure total lipase activity,
plasma samples were incubated with 10 % Intralipid / [3H]
TG emulsion (Hospira) as a substrate and human serum as the source of ApoC - III (24).
Hypertriglyceridemia is an independent risk factor for cardiovascular disease, and
plasma triglycerides (
TGs) correlate strongly with
plasma apolipoprotein C - III (ApoC - III) levels.
(C) Lipoprotein
TG profiles in
plasma from pooled, fasted iLpldf mice treated with tamoxifen to induce inactivation of LPL expression (iLpldf)(n = 3 pooled mice / group).
Lipoprotein fractions were prepared from
plasma by sequential ultracentrifugation.18 Cholesterol and
TG contents were determined by automated chemical analysis.
Hypertriglyceridemia, defined as a fasting
plasma Triglyceride (
TG) concentration of more than 150 mg / dL, is a common condition in the United States.
The mice without an adipocyte clock and shifted feeding rhythms developed obesity, as well as changes in the relative amounts of various
TGs stored in adipocytes, circulating in the
plasma, and present in hypothalamic neurons associated with energy balance (31).