Salivary ceruloplasmin ferroxidase & oxidase activities in celiac patients

Abstract

The aim of the current study was to evaluate salivary ferroxidase ceruloplasmin activities in celiac patients with different histopathological severity. This study included 75 celiac patients with different mean age (18.68 ± 11.13) year, who had positive screen for celiac antibodies, and who had gastrointestinal symptoms. In order to simplify the comparison with the healthy control group, celiac patients were divided into two groups according to their histopathological severity: severe (marsh IIIa, b, c) & less severe (marsh 0, I). All these patients have been evaluating for salivary ceruloplasmin (Cp) concentration and Cp ferroxidase activities. To confirm the presence of the enzymatic activity of this protein, polyacrylamide gel electrophoresis was carried out and then stained for Cp ferroxidase, as well as for Cp oxidase activity. Furthermore, the concentrations of salivary total protein, albumin, and globulin were measured in the studied groups. A significant increase (p<0.05) in salivary concentration of ceruloplasmin was found in all above mentioned patients groups in comparison to that of the control group, except for total villous atrophy (marsh IIIc) patients subgroup. Salivary Cp ferroxidase activity revealed statistically significant decrease among the patient groups as well as between them and the control group. The result of salivary total protein and globulin showed presence a significant increase (p<0.05) in comparison to that of the control group. Meanwhile albumin levels was found to increase non-significantly (p=0.186).

Keywords: celiac disease; ceruloplasmin; electrophoresis; mucosal histopathological damage; saliva.

Figure 1

Marsh distribution among study groups.

Figure 2

Conventional polyacrylamide gel electrophoresis (PAGE) 7.5%, using Tris- glycine buffer, pH 8.9 as electrode buffer. Electrophoresis was carried out for 3 hours at 4°C by using a constant current of 40 mA & voltage of 15 v/cm. The gel was stained for protein. Concentrated pooled saliva samples(with a protein concentration of 1.8 mg/ml ) were applied as follows: 1 p ooled c rude s aliva (marsh I IIa), 2 p ooled c rude saliva(r IIIb), 3 pooled crude saliva (marsh IIIc), 4 pooled crude saliva (control), 5 pooled crude saliva (mash 0), 6 pooled crude saliva (marsh I).

Figure 3

Discontinuous Polyacrylamide tubular Gel-Electrophoresis. (Lammeli system using 4% stacking gel and 7.5% separating gel) was carried out using Pharmacia Gel Electrophoresis Apparatus GE-2/4 LS. And Tris- glycine buffer, pH 8.2 as electrode buffer. Electrophoresis was carried out for 2.5 hours at 4°C with a constant current of 38 mA & a voltage of 15 v/cm. The gel was stained for CP ferroxidase activity. The applied concentrated samples (4.2 mg/ml) were as follows from up to down: pooled crude saliva (marsh IIIa), pooled crude saliva (marsh IIIb), pooled crude saliva (marsh IIIc), pooled crude saliva (marsh I), pooled crude saliva (marsh 0), and pooled crude saliva (control). (a) In the presence of sodium azide and (b) in the absence of sodium azide.

Figure 4

Conventional polyacrylamide slab gel electrophoresis (PAGE) 7.5%, using Tris- glycine buffer, pH 8.9 as electrode buffer. Electrophoresis was carried out for 3 hours at 4°C by using a constant current of 40 mA & voltage of 15 v/cm. The gel was stained for Cp oxidase activity. The applied samples (1.8 mg/ml) were as follows: 1- pooled crude saliva (control), 2 - pooled crude saliva (marsh IIIa), 3 - pooled crude saliva (marsh IIIb), 4 - pooled crude saliva (marsh IIIc), 5- pooled crude saliva (marsh 0), 6- pooled crude saliva (marsh I). (a) In the presence of sodium azide and (b) in the absence of sodium azide