In 1995, the
Male Wistar rats weighing
In accordance with the model reference utilized [
Rats of the restrained group were only fastened to the operating table and were not injected with any chemicals or solution. Rats of the blank-control group were neither fastened to the operating table nor injected with any chemicals or solution; these rats were not interfered with in any way.
The first-dimensional chromatographic column of Strong Cation Exchange (SCX) was Agilent ZORBAX Bio-SCX Series II (50 × 0.8 mm, 3
Nanoliter analytical RPLC’s second-dimensional column: the C18 column was Agilent ZORBAX SB (150 × 0.5 mm, 5
SPSS 11.0 statistical software was used to process the data, which was expressed as mean ± SD. One-way ANOVA was used in the simultaneous comparison of the 3 groups. LSD method was used for instances of equal variances, and Dunnett T3 was used for any unequal variances. A result of
In order to identify the different peaks and those enzymatic hydrolysates displaying varying peak areas of common peak shapes, the peak area of the enzymatic hydrolysates from the asthma model group was compared with the two other groups (as shown in Figures
Areas of serum peptide peaks comparison in 3 groups (mean ± SD).
Number | Ret. time (min) | A (blank-control group) | B (restrained group) | C (asthma model group) |
---|---|---|---|---|
1 | 77.489 | 177.3077 ± 4.8170 | 64.64110 ± 7.1792(2) | |
2 | 78.418 | 86.82484 ± 3.2402(2) | ||
3 | 78.804 | 65.57249 ± 48.1971 | ||
4 | 79.398 | 37.60763 ± 8.8873(2) | 230.9545 ± 124.0502 | |
5 | 80.533 | 61.07169 ± 7.8572(2) | ||
6 | 81.154 | 225.9217 ± 173.4557 | 120.6287 ± 4.3967 | 254.2249 ± 136.8851 |
7 | 81.824 | 128.3332 ± 41.5655(1) | 55.8214 ± 7.7610 | |
8 | 85.358 | 1591.588 ± 1014.9047 | 614.3952 ± 162.0699(1) | 1242.374 ± 320.6863 |
9 | 86.992 | 154.6185 ± 53.7288 | 124.6411 ± 15.6364 | 123.3418 ± 47.1595 |
10 | 89.047 | 76.36887 ± 16.2800 | 68.38598 ± 52.6746 | 89.51789 ± 29.1391 |
11 | 92.066 | 1056.652 ± 406.739 | 1033.819 ± 412.7315 | 1017.987 ± 794.5467 |
12 | 93.113 | 208.6571 ± 127.1982 | 49.48719 ± 6.9568(1) | 174.6457 ± 67.7304(3) |
13 | 94.731 | 48.7882 ± 7.4086(4) | ||
14 | 101.436 | 95.77694 ± 15.2855 | 88.06084 ± 14.1367 | 107.5445 ± 20.0045 |
15 | 103.410 | 596.3253 ± 442.7393 | 267.3705 ± 61.2447 | 430.9182 ± 107.1599 |
The peaks of peptides in the serum of 3 groups. (A) Blank-control group, (B) restrained group, and (C) asthma model group.
The peaks of peptides in the serum of blank-control group.
The peaks of peptides in the serum of restrained group.
The peaks of peptides in the serum of asthma model group.
(1) Three groups share 8 common peaks: peak number 6 (81.154 min), peak number 8 (85.358 min), peak number 9 (86.992 min), peak number 10 (89.047 min), peak number 11 (92.066 min), peak number 12 (93.113 min), peak number 14 (101.436 min), and peak number 15 (103.410 min).
(2) Comparing the restrained group with the blank-control group, 4 new peaks appeared: peak number 2 (78.418 min), peak number 4 (79.398 min), peak number 5 (80.533 min), and peak number 7 (81.824 min). This suggests that there might be a correlation between these 4 polypeptides and the binding-induced stress stimuli. Among the 9 common peaks, peak number 1 displayed a significant difference in the peak area (
(3) Comparing the asthma model group with the restrained group, 1 new specific peak appeared: peak number 13 (94.731 min). Three peaks lost their specificity: peak number 1 (77.489 min), peak number 2 (78.418 min), and peak number 5 (80.533 min), signifying that these polypeptides are related to the pathogenesis of allergic asthma.
There were a total of 10 common peaks, among which peak number 12 displayed significant variation on the peak area (
The deciphering of the human genetic code opened the next chapter in the scientific study of human beings; nonetheless, current knowledge of the human genome is still unable to explain the mechanism of the genome’s compiled protein [
Nanoscaled two-dimensional capillary liquid chromatography is characterized by a straightforward operational procedure, efficient separation effect, high resolution, stability, and repeatability; consequently, it currently has wide application in the separation, purification, and preparation of polypeptides. As the flow rate is controlled at the nL/min level, the sample volume is significantly reduced, the mass spectrometry data acquisition time is extended, and the detection sensitivity is improved, which makes it more suitable for the separation and identification of peptide mixtures with only a few samples. Nano-2D-LC was thus used in this research to detect the low-abundance protein in the serum of rats with allergic asthma. Results indicated that the peptide peak at 79.398 min was the peak of asthma-related protein. Further mass spectrometric identification of retention-time effluent materials may lead to the identification of several enzymatic hydrolysate-related proteins associated with the onset of asthma in the serum.
Yuan and his team [
At present, the study of asthma’s mechanism is still a major obstacle in the medical field; compared to tumor research, it does not contain easily available target cells. The pathogenesis mechanism of asthma is complex. In etiology, it is closely related to genetics and the environment. In pathogenesis, it is related to the immune system, nervous system, and their mutual interactions. Finally, it is related to eosinophils, neutrophils, mast cells, and lymphocytes in pathophysiology. Through an analysis of the available literature, we found that asthma proteins are mostly associated with redox, airway’s remodeling, anti-inflammation, and signaling pathways. Some types of proteins can be important biomarkers and therapeutic targets for asthma. Today, there are still many issues associated with the current application of proteomics research on bronchial asthma. However, through the combination of capillary electrophoresis, chromatography, and mass spectrometry and with the recent rapid development of protein Chip-TOF-MS technology, it is now possible to advance the study of asthma’s biological mechanism from the perspective of proteomics and obtain new clinical diagnoses and treatment targets.
The authors have no financial disclosures or conflicts of interest to declare.
This study was supported by Henan Shao’s Acupuncture School Inheritance Studio & Gansu University of Chinese Medicine Workstation Project of State Administration of Traditional Chinese Medicine (no. LP0116036-z8), the Basic Research Foundation of Higher Education Institutions of Gansu Provincial Department of Finance (no. 2011-4), and the Introduction of Talent Scientific Research Start-Up Foundation of Gansu University of Chinese Medicine (no. 2012-7).