Methodological Studies Of Serum Lipoprotein Electrophoretogram Detection: Basic Validation. Particle Properties Of Serum Lipoproteins And Subfractions In Normolipidemic Humen

Numerous studies have shown a closed correlation between dyslipid-emia and cardiocerebral vascular diseases. However, there are lots of questions not to be figured out yet, chiefly attributed to limited recognition about the association between whole lipoprotein particles and their functions, and the neglect of the importance of their dynamic balance. The problems of synchronous separation and uniform reference have not been solved effectively, and clinical detection has failed to express the state of their dynamic balance.We had estabolished the method of serum lipoprotein electrophore-togram (SLPG) which could synchronously separate serum lipoproteins into 9~11 bands including albumin-nonesterified fatty acids (AL-NEFA), five subfractions of alpha-lipoproteins ( a Lpi-s), beta-lipoproteins (?Lp), inter-beta-lipoproteins (int?Lp), two subfractions of pre-beta-lipoproteins (pre P Lp1-2) and chylomicrons (CM). This method applied the quantitative analysis of unique reference and expressed the state of serum lipoprotein dynamic balance completely and concretely, which effective performance is the only one in the similar worldwide field. Previous studies have demonstrated its obvious clinical significance and applying prospects. Objectives:1. To investigate whether SLPG is scientific or not, verify the connotation of each electrophoretic peak in SLPG, and accomplish the basic validation of SLPG dectection.2. To observe the physical and chemical properties of serum lipoproteins and subfractions involving particle morphology, size and the components of apolipoproteins (apo) and cholesterol (Chol).3. To observe the particle sizes and distributions of normolipidemic serum lipoproteins and subfractions.4. To investigate the validity and confidence of small and dense low density lipoproteins (sLDL). Materials and Methods:1. Compound sera came from 134 adult blood samples. The concentrations of routine blood lipids including triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were all in the recommended suitable or borderline ranges.2. Serum lipoproteins were separated by the method of double gradient polyacrylamide gel electrophoresis (DGPAGE). They were collected after elution from electrophoretic gels respectively.3. The morphologies of serum lipoproteins and subfractions were observed applying transmission electron microscopy and negative staining technique. Lipoprotein particles in the electron micrographs were counted and measured by the softwares of Photoshop 7.0 and Image Pro 4.5.4. The amounts of apo in lipoprotein solutions collected after elution were detected by the immuno-turbidimetry, and those of Chol by the enzymatic method.Results:1. The morphologies of normolipidemic serum lipoproteins were predominant spheroidal particles presented by electron microscopy.2. The spheroidal particle sizes of serum lipoproteins and subfractions changed with a certain sequence. The spheroidal particle diameter of AL-NEFA was smallest (3.15+0.40nm), a Lp subfractions from 5.85 + 0.81nm to 13.32?.02nm, ?Lp 25.75?.40nm, int週p 34.83?.74nm, and pre週p subfractions 39.82?.09nm and 39.2612.57nm. They had significant differences (P<0.05) between each other except for the last ones. The size distributions of spheroidal particles presented single peak in AL-NEFA and a Lp subfractions, and double peaks in the others. The particles with a diameter of fewer than 25.5nm occupied 57.66% in the ?Lp spheroidal ones.3. The weight ratios of apo and Chol in various lipoproteins and subfractions changed with a certain sequence.(1) The ratio of apo A I Chol (WAV) was the highest in AL-NEFA (4.80 0.54). the means of that were in the range of 4.21-1.45 in a Lp subfractionsand gradually decreased with a sequence of their electrophoretic mobilities. They had significant differences (P<0.05) between each other except for a Lp3 and a Lp4.(2) The ratios of apo B/Chol (WAV) in P Lp, int P Lp and pre P Lpi (0.85 ?0.22, 0.96?.30 and 1.08 +...