Figure 1.
Establishment of Leprdb/dbSmp30Y/− mice.
(A) Generation of Leprdb/dbSmp30Y/− mice. F1 hybrid mice (in green boxes) were crossed to produce the experimental Leprdb/+Smp30Y/+, Leprdb/+Smp30Y/−, Leprdb/dbSmp30Y/+ and Leprdb/dbSmp30Y/− mice (in red boxes). (B) Appearance of Leprdb/dbSmp30Y/− mice at 24 weeks of age. (C) Western blot analysis of SMP30 protein levels in livers from each experimental group at 24 weeks of age. (D) Vitamin C concentration in livers from each experimental group of mice at 24 weeks of age. Values are given as means ± SEM of five animals.
Table 1.
Primer sequences for use in real-time quantitative RT-PCR.
Figure 2.
Decrease in hepatic SMP30 levels in Leprdb/db mice.
(A) SMP30 mRNA and (B) protein levels in livers from Leprdb/+Smp30Y/+ and Leprdb/dbSmp30Y/+ mice. Values are given as means ± SEM of five animals. †P<0.01.
Table 2.
Physiological, blood and biochemical parameters in four experimetal groups of mice.
Figure 3.
Increase in small dense LDL-cho and decrease in HDL-cho in plasma from Leprdb/dbSmp30Y/− mice.
(A) HPLC lipoprotein profile with cholesterol reagent and (B–F) cholesterol content in four subfractions according particle size, VLDL; Fr. 3–7, LDL; Fr. 8–11, small dense LDL; Fr. 12–13, HDL; Fr. 14–20 and LDL/HDL ratio in the four experimental groups. Blue dashed line and lane 1, Leprdb/+Smp30Y/+ mice; Red dashed line and lane 2, Leprdb/+Smp30Y/− mice; Blue line and lane 3, Leprdb/dbSmp30Y/+ mice; Red line and lane 4; Leprdb/dbSmp30Y/−mice. Values are given as means ± SEM of five animals. †P<0.01 and §P<0.001 versus Leprdb/+Smp30Y/+, ¶P<0.001 versus Leprdb/+Smp30Y/−, **P<0.05 and ††P<0.001 versus Leprdb/dbSmp30Y/+.
Figure 4.
Increase in TBARS in livers from Leprdb/dbSmp30Y/− mice.
(A) Triglyceride, (B) Cholesterol, (C) Phospholipid and (D) TBARS content in livers from the four experimental groups. Lane 1, Leprdb/+Smp30Y/+ mice. Lane 2, Leprdb/+Smp30Y/− mice. Lane 3, Leprdb/dbSmp30Y/+ mice. Lane 4, Leprdb/dbSmp30Y/− mice. †P<0.01 and §P<0.001 versus Leprdb/+Smp30Y/+, ¶P<0.001 versus Leprdb/+Smp30Y/−, **P<0.05 and ††P<0.001 versus Leprdb/dbSmp30Y/+. Values are given as means ± SEM of five animals.
Figure 5.
Increased steatosis, inflammation and oxidative stress in liver sections from Leprdb/dbSmp30Y/− mice.
Representative images of (A) hematoxylin/eosin staining and (B) 4-HNE immunostaining in liver sections from Leprdb/+Smp30Y/+, Leprdb/+Smp30Y/−, Leprdb/dbSmp30Y/+ and Leprdb/dbSmp30Y/− mice. Scale bar is 100 µm. *P<0.05, †P<0.01 and §P<0.001 versus Leprdb/+Smp30Y/+, #P<0.05, ‡P<0.01 and ¶P<0.001 versus Leprdb/+Smp30Y/−, **P<0.05 versus Leprdb/dbSmp30Y/+. Values are given as mean ± SEM of five animals.
Figure 6.
Altered hepatic expression of lipid/lipoprotein metabolism-related genes in Leprdb/dbSmp30Y/− mice.
Gene expression levels of (A) ACC, (B) FAS, (C) SREBP1c, (D) SREBP2, (E) HMGCoAR, (F) PPARα, (G) MCAD, (H) MTP, (I) ApoB100, (J) LDLR (K) VLDLR, and (L) CD36 in the livers. Lane 1: Leprdb/+Smp30Y/+, lane 2: Leprdb/+Smp30Y/−, lane 3: Leprdb/dbSmp30Y/+ and lane 4: Leprdb/dbSmp30 Y/− mice. mRNA for each gene was measured using real time RT-PCR and normalized to β-actin. The values from Leprdb/+ Smp30Y/+ mice were assigned a relative value of 1.0. Values are given as means ± SEM of five animals. *P<0.05, †P<0.01 and §P<0.001 versus Leprdb/+Smp30Y/+, #P<0.05, ‡P<0.01 and ¶P<0.001 versus Leprdb/+Smp30Y/−, **P<0.05 versus Leprdb/dbSmp30Y/+.
Figure 7.
Increase in endoplasmic reticulum (ER) stress in livers from Leprdb/dbSmp30Y/− mice.
(A) Spliced XBP1 mRNA expression, (B) Phosphorylated eIF2α protein and (C) CHOP protein levels in livers from the four experimental groups. Values are given as means ± SEM of five animals. *P<0.05, †P<0.01 and §P<0.001 versus Leprdb/+Smp30Y/+, #P<0.05 versus Leprdb/+Smp30Y/−, **P<0.05 versus Leprdb/dbSmp30Y/+.