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Altered Expression Levels of MMP1, MMP9, MMP12, TIMP1, and IL-1β as a Risk Factor for the Elevated IOP and Optic Nerve Head Damage in the Primary Open-Angle Glaucoma Patients.

Markiewicz L, Pytel D, Mucha B, Szymanek K, Szaflik J, Szaflik JP, Majsterek I - Biomed Res Int (2015)

Bottom Line: In vivo promoter activity assays were carried on HTM cells using dual luciferase assay.All studied subjects underwent ophthalmic examination, including BCVA, intraocular pressure, slit-lamp examination, gonioscopy, HRT, and OCT scans.Allele -1607 1G of MMP1 gene possesses only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possesses only 21,86% of the -1562 T allele.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-647 Lodz, Poland.

ABSTRACT
The aim of presented work was to analyze the impact of particular polymorphic changes in the promoter regions of the -1607 1G/2G MMP1, -1562 C/T MMP9, -82 A/G MMP12, -511 C/T IL-1β, and 372 T/C TIMP1 genes on their expression level in POAG patients. Blood and aqueous humor samples acquired from 50 patients with POAG and 50 control subjects were used for QPCR and protein levels analysis by ELISA. In vivo promoter activity assays were carried on HTM cells using dual luciferase assay. All studied subjects underwent ophthalmic examination, including BCVA, intraocular pressure, slit-lamp examination, gonioscopy, HRT, and OCT scans. Patients with POAG are characterized by an increased mRNA expression of MMP1, MMP9, MMP12, and IL-1β genes as compared to the control group (P < 0.001). Aqueous humor acquired from patients with POAG displayed increased protein expression of MMP1, MMP9, MMP12, and IL-1β compared to the control group (P < 0.001). Allele -1607 1G of MMP1 gene possesses only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possesses only 21,86% of the -1562 T allele. Increased expression levels of metalloproteinases can be considered as a risk factor for the development of POAG.

No MeSH data available.


Related in: MedlinePlus

MMPs promoter activity. Plasmid constructs harboring: MMP1 2G: insertion of additional guanine in position -1607 (2G) in MMP1 gene (pGL 4.11 MMP1 2G); MMP9 T: change in position -1562 from C to T in MMP9 gene. pGL 4.11 nc: negative control (promoterless); pGL 4.13: positive control (CMV promoter). Data presented as relative luciferase units RLU calculated as FLA/RLA of the sample. Error bars represent standard deviations (SD). ∗∗∗P < 0.001. Data from three independent experiments with independent transfections (n = 3).
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fig3: MMPs promoter activity. Plasmid constructs harboring: MMP1 2G: insertion of additional guanine in position -1607 (2G) in MMP1 gene (pGL 4.11 MMP1 2G); MMP9 T: change in position -1562 from C to T in MMP9 gene. pGL 4.11 nc: negative control (promoterless); pGL 4.13: positive control (CMV promoter). Data presented as relative luciferase units RLU calculated as FLA/RLA of the sample. Error bars represent standard deviations (SD). ∗∗∗P < 0.001. Data from three independent experiments with independent transfections (n = 3).

Mentions: We utilized dual luciferase assay to assess the influence of the -1607 1G to -1607 2G allele change on the level of MMP1 and -1562 C to -1562 T allele on MMP9 expression. Results showed an increase in relative luciferase units (RLU) for the construct harboring allele -1607 2G of MMP1 gene compared to -1607 1G (15.21 RLU and 6.52 RLU resp.) and for the allele -1562 T of MMP9 gene compared to -1562 C (20.69 RLU and 4.52 RLU, resp.) (Figure 3). The allele -1607 1G of MMP1 gene possess only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possess only 21,86% of the -1562 T allele transcriptional activity.


Altered Expression Levels of MMP1, MMP9, MMP12, TIMP1, and IL-1β as a Risk Factor for the Elevated IOP and Optic Nerve Head Damage in the Primary Open-Angle Glaucoma Patients.

Markiewicz L, Pytel D, Mucha B, Szymanek K, Szaflik J, Szaflik JP, Majsterek I - Biomed Res Int (2015)

MMPs promoter activity. Plasmid constructs harboring: MMP1 2G: insertion of additional guanine in position -1607 (2G) in MMP1 gene (pGL 4.11 MMP1 2G); MMP9 T: change in position -1562 from C to T in MMP9 gene. pGL 4.11 nc: negative control (promoterless); pGL 4.13: positive control (CMV promoter). Data presented as relative luciferase units RLU calculated as FLA/RLA of the sample. Error bars represent standard deviations (SD). ∗∗∗P < 0.001. Data from three independent experiments with independent transfections (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4442285&req=5

fig3: MMPs promoter activity. Plasmid constructs harboring: MMP1 2G: insertion of additional guanine in position -1607 (2G) in MMP1 gene (pGL 4.11 MMP1 2G); MMP9 T: change in position -1562 from C to T in MMP9 gene. pGL 4.11 nc: negative control (promoterless); pGL 4.13: positive control (CMV promoter). Data presented as relative luciferase units RLU calculated as FLA/RLA of the sample. Error bars represent standard deviations (SD). ∗∗∗P < 0.001. Data from three independent experiments with independent transfections (n = 3).
Mentions: We utilized dual luciferase assay to assess the influence of the -1607 1G to -1607 2G allele change on the level of MMP1 and -1562 C to -1562 T allele on MMP9 expression. Results showed an increase in relative luciferase units (RLU) for the construct harboring allele -1607 2G of MMP1 gene compared to -1607 1G (15.21 RLU and 6.52 RLU resp.) and for the allele -1562 T of MMP9 gene compared to -1562 C (20.69 RLU and 4.52 RLU, resp.) (Figure 3). The allele -1607 1G of MMP1 gene possess only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possess only 21,86% of the -1562 T allele transcriptional activity.

Bottom Line: In vivo promoter activity assays were carried on HTM cells using dual luciferase assay.All studied subjects underwent ophthalmic examination, including BCVA, intraocular pressure, slit-lamp examination, gonioscopy, HRT, and OCT scans.Allele -1607 1G of MMP1 gene possesses only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possesses only 21,86% of the -1562 T allele.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-647 Lodz, Poland.

ABSTRACT
The aim of presented work was to analyze the impact of particular polymorphic changes in the promoter regions of the -1607 1G/2G MMP1, -1562 C/T MMP9, -82 A/G MMP12, -511 C/T IL-1β, and 372 T/C TIMP1 genes on their expression level in POAG patients. Blood and aqueous humor samples acquired from 50 patients with POAG and 50 control subjects were used for QPCR and protein levels analysis by ELISA. In vivo promoter activity assays were carried on HTM cells using dual luciferase assay. All studied subjects underwent ophthalmic examination, including BCVA, intraocular pressure, slit-lamp examination, gonioscopy, HRT, and OCT scans. Patients with POAG are characterized by an increased mRNA expression of MMP1, MMP9, MMP12, and IL-1β genes as compared to the control group (P < 0.001). Aqueous humor acquired from patients with POAG displayed increased protein expression of MMP1, MMP9, MMP12, and IL-1β compared to the control group (P < 0.001). Allele -1607 1G of MMP1 gene possesses only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possesses only 21,86% of the -1562 T allele. Increased expression levels of metalloproteinases can be considered as a risk factor for the development of POAG.

No MeSH data available.


Related in: MedlinePlus