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Comparative study of serum proteomes in Legg-Calve-Perthes disease.

Liu R, Fan L, Yin L, Wang K, Miao W, Song Q, Dang X, Gao H, Bai C - BMC Musculoskelet Disord (2015)

Bottom Line: Age- and sex-matched serum samples from 10 control subjects and 10 patients with LCPD were compared using the isobaric tags for relative and absolute quantification (iTRAQ) technique.Gene ontology analyses, KEGG pathway and functional network analyses were performed.The alpha-1-acid glycoprotein 1 and haptoglobin increases, and apolipoprotein E and S100-A8 decreases were confirmed by western blot.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic, the Second Hospital Affilicated to Medical College, Xi'an Jiaotong University, No.157, Xiwu Road, Xi'an, Shaanxi, 710004, P. R China. liuryu@126.com.

ABSTRACT

Background: Legg-Calve-Perthes Disease (LCPD) is an idiopathic osteonecrosis of the developing femoral head complicated by pain and disability of the hip joint. To date, the pathological mechanisms of LCPD are not well-known. This study screened the changes in serum protein expression in patients with LCPD.

Methods: Age- and sex-matched serum samples from 10 control subjects and 10 patients with LCPD were compared using the isobaric tags for relative and absolute quantification (iTRAQ) technique. Gene ontology analyses, KEGG pathway and functional network analyses were performed. Proteins of interest with large differences in expression, S100-A8, alpha-1-acid glycoprotein 1, haptoglobin and apolipoprotein E, were compared by western blotting.

Results: The disease/control ratios showed 26 proteins were significantly differentially expressed (all p < 0.05). Including higher abundances of complement factor H (1.44), complement C4-B (1.45), isocitrate dehydrogenase [NAD] subunit alpha (2.7) alpha-1-acid glycoprotein 1 (1.87), heptoglobin (1.53) and Ig lambda-2 chain C regions (1.46), and lower levels of apolipoprotein E (0.50), apolipoprotein F (0.60), apolipoprotein C-III (0.69), S100-A8 (0.73), S100-A9 (0.75) and prothrombin (0.77) in LCPD than in controls. The alpha-1-acid glycoprotein 1 and haptoglobin increases, and apolipoprotein E and S100-A8 decreases were confirmed by western blot. KEGG pathway analysis revealed these proteins were related to the complement and coagulation cascades, Staphylococcus aureus infection, PPAR signaling, fat digestion and absorption, and vitamin digestion and absorption. Functional network analysis suggested that the proteins were involved in lipid regulation.

Conclusions: The complement and coagulation cascades, and abnormal lipid metabolism may be involved in the pathogenesis of LCPD.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of four differentially expressed proteins. Relative intensities of the positively identified proteins are shown by the histograms. Patients with Legg-Calve-Pathes disease (LCPD) displayed higher levels of a orosomucoid 1 (ORM1) and b haptoglobin (HP), and lower levels of c apolioprotein E (APOE) and d S100-A8 than healthy controls (HC). The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. *indicates significant difference from the healthy volunteer group, p < 0.05
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Fig4: Western blot analysis of four differentially expressed proteins. Relative intensities of the positively identified proteins are shown by the histograms. Patients with Legg-Calve-Pathes disease (LCPD) displayed higher levels of a orosomucoid 1 (ORM1) and b haptoglobin (HP), and lower levels of c apolioprotein E (APOE) and d S100-A8 than healthy controls (HC). The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. *indicates significant difference from the healthy volunteer group, p < 0.05

Mentions: Western blot analysis of four differentially expressed proteins (S100-A8, haptoglobin, alpha-1-acid glycoprotein 1 and apolipoprotein E) was used to verify our mass spectrometry results. They were selected for investigation based on their interesting biological functions and high fold-changes, as well as the availability of commercial antibodies. ORM1 and HP were increased in patients with LCPD, APOE and S100-A8 were decreased in patients with LCPD (shown in Fig. 4). Thus, western blotting was consistent with the results of the mass spectrometry analysis.Fig. 4


Comparative study of serum proteomes in Legg-Calve-Perthes disease.

Liu R, Fan L, Yin L, Wang K, Miao W, Song Q, Dang X, Gao H, Bai C - BMC Musculoskelet Disord (2015)

Western blot analysis of four differentially expressed proteins. Relative intensities of the positively identified proteins are shown by the histograms. Patients with Legg-Calve-Pathes disease (LCPD) displayed higher levels of a orosomucoid 1 (ORM1) and b haptoglobin (HP), and lower levels of c apolioprotein E (APOE) and d S100-A8 than healthy controls (HC). The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. *indicates significant difference from the healthy volunteer group, p < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4595068&req=5

Fig4: Western blot analysis of four differentially expressed proteins. Relative intensities of the positively identified proteins are shown by the histograms. Patients with Legg-Calve-Pathes disease (LCPD) displayed higher levels of a orosomucoid 1 (ORM1) and b haptoglobin (HP), and lower levels of c apolioprotein E (APOE) and d S100-A8 than healthy controls (HC). The relative density was calculated by dividing the density of matched spot by the density of all the matched spots in the respective gel. *indicates significant difference from the healthy volunteer group, p < 0.05
Mentions: Western blot analysis of four differentially expressed proteins (S100-A8, haptoglobin, alpha-1-acid glycoprotein 1 and apolipoprotein E) was used to verify our mass spectrometry results. They were selected for investigation based on their interesting biological functions and high fold-changes, as well as the availability of commercial antibodies. ORM1 and HP were increased in patients with LCPD, APOE and S100-A8 were decreased in patients with LCPD (shown in Fig. 4). Thus, western blotting was consistent with the results of the mass spectrometry analysis.Fig. 4

Bottom Line: Age- and sex-matched serum samples from 10 control subjects and 10 patients with LCPD were compared using the isobaric tags for relative and absolute quantification (iTRAQ) technique.Gene ontology analyses, KEGG pathway and functional network analyses were performed.The alpha-1-acid glycoprotein 1 and haptoglobin increases, and apolipoprotein E and S100-A8 decreases were confirmed by western blot.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic, the Second Hospital Affilicated to Medical College, Xi'an Jiaotong University, No.157, Xiwu Road, Xi'an, Shaanxi, 710004, P. R China. liuryu@126.com.

ABSTRACT

Background: Legg-Calve-Perthes Disease (LCPD) is an idiopathic osteonecrosis of the developing femoral head complicated by pain and disability of the hip joint. To date, the pathological mechanisms of LCPD are not well-known. This study screened the changes in serum protein expression in patients with LCPD.

Methods: Age- and sex-matched serum samples from 10 control subjects and 10 patients with LCPD were compared using the isobaric tags for relative and absolute quantification (iTRAQ) technique. Gene ontology analyses, KEGG pathway and functional network analyses were performed. Proteins of interest with large differences in expression, S100-A8, alpha-1-acid glycoprotein 1, haptoglobin and apolipoprotein E, were compared by western blotting.

Results: The disease/control ratios showed 26 proteins were significantly differentially expressed (all p < 0.05). Including higher abundances of complement factor H (1.44), complement C4-B (1.45), isocitrate dehydrogenase [NAD] subunit alpha (2.7) alpha-1-acid glycoprotein 1 (1.87), heptoglobin (1.53) and Ig lambda-2 chain C regions (1.46), and lower levels of apolipoprotein E (0.50), apolipoprotein F (0.60), apolipoprotein C-III (0.69), S100-A8 (0.73), S100-A9 (0.75) and prothrombin (0.77) in LCPD than in controls. The alpha-1-acid glycoprotein 1 and haptoglobin increases, and apolipoprotein E and S100-A8 decreases were confirmed by western blot. KEGG pathway analysis revealed these proteins were related to the complement and coagulation cascades, Staphylococcus aureus infection, PPAR signaling, fat digestion and absorption, and vitamin digestion and absorption. Functional network analysis suggested that the proteins were involved in lipid regulation.

Conclusions: The complement and coagulation cascades, and abnormal lipid metabolism may be involved in the pathogenesis of LCPD.

No MeSH data available.


Related in: MedlinePlus