Role of electronegative LDL in coronary hyperpermeability

陰電性低密度脂蛋白在冠狀動脈高通透性所扮演的角色

Abstract
 The relationship between high cholesterol level and cardiovascular disease has been recognized, but in recent years, some studies found that not all kinds of high level of “bad cholesterol”, whose formal name is “low density lipoprotein (LDL)” , will lead to cardiovascular occlusion, in fact, electronegative LDL  (L5) is the culprit of inducing apoptosis of epithelial cells and myocytes. Therefore, this study will focus on exploring the relationship between L5 and coronary epithelial cells.
 This study also investigate the relationship between spinal cord injury and LDL. We constructed two groups of animal model – the surgery group (damage the spinal nerve of mice by surgery) and the control group, and then we took out their hearts and analyzed the changes of the proteins in the hearts. Finally, we found that the level of the expression of LDL in spinal cord injury group increased, and this result also indicate that there is a relationship between coronary hyperpermeability and spinal cord injury.

Materials

1. Animal sample
2. sodium dodecyl sulfate polyacrylamide gel
3. isopropanol
4. BCA protein assay kit
5. transfer kit
6. protein marker
7. buffer
1. running buffer
2. transfer buffer
3. blocking buffer
4. washing buffer：TBST
8. primary antibody: SirT1, beta tubulin, Lox-1
9. secondary antibody: mouse, rabbit
10. oxidizing reagent＋enhanced luminal reagent

Method
Bradford protein assay
Western blot

Result

Figure 1. BCA standard curve

Figure 2. The gel has 300 seconds of exposure. The samples of surgery group and the control group are on different gel, and “m” means “marker”.

Figure 3. The gel has 600 seconds of exposure. There are both samples of surgery group and control group on one gel.

Discussion

1. Sirtulin 1
    In the case of  300 seconds of exposure,  we found that the level of the expression of SirT1 in surgery group is not obvious, but in the case of 600 seconds of exposure, there is no obvious difference between surgery group and control group.
    Conjecturing that because the surgery group sample and the control group sample in the case of 300 seconds of exposure are on different gels, that cause extra variables, so the comparability decline.
2. Beta tubulin
    Beta tubulin is one of the important part of microtubule, so we used it to be our “internal control”, and the usefulness of internal control is making sure that the amount of protein in every sample are same, so if the sample is from cell or tissue, the expression of beta tubulin would be very obvious.
    However, neither in the case of 300 nor 600 seconds of exposure, the expression of beta tubulin aren’t obvious,  so in this study, beta tubulin can’t be our internal control.
3. Lox-1（lectin-type oxidized LDL receptor 1）
    In Figure 3, we can easily find that the level of the expression of Lox-1 in surgery group is more than control group, whether color depth or the width of the dark band.
   This result is contrary to the conclusion of D Cardús in 1992, they considered that spinal cord injury would not affect the amount of LDL and HDL.
    Perhaps there are species variation between animal model and human, whatever, the result of our study may provide some improvement to follow-up researchers.