Journal, genetic engineering & biotechnology · 2025
Profiling of proteases involved in SARS-CoV-2 pathogenesis in human saliva: Influence of age and gender.
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Abdellatif Mentalla Motaz, El Amir Azza, Shalaby Said, Kirschfink Michael, Nadeem Rola, Abd-Elshafy Dina Nadeem, Bahgat Mahmoud Mohamed
Аннотация
BACKGROUND: SARS-CoV-2 enters human cells via angiotensin-converting enzyme 2 (ACE2). In silico experiments also demonstrated possible SARS-CoV-2 binding to dipeptidyl peptidase 4 (DPP-4) during cell entry. Moreover, matrix metalloproteinase 2 (MMP-2) is involved in the SARS-CoV-2 cell fusion process.
AIMS: To study expression and activity profiles of implicated enzymes in SARS-CoV-2 infection in various age groups of both genders and discuss the possible impact of this on COVID-19 outcome.
METHODS: Saliva from different age groups was used to profile MMP-2 and gelatinase enzymes by zymography. Enzyme activities resulting from the cleavage of the H-Arg-Pro-pNA substrate were measured using a microplate reader. ACE2 transcription and translation were quantified using qRT-PCR and ELISA. Statistical analysis was used to identify the presence of significant differences.
RESULTS: Zymography demonstrated different gelatinase activities in a range of ∼180-30 kDa and showed distinct bands at ∼96, 72, 66, 60, and 40 kDa. Comparing MMP-2, DPP-4, DPP-4/ACE2 activities, ACE2 protein, and mRNA levels between genders showed p-values of 0.0142, 0.1056, 0.1859, 0.6411, and 0.4797, whereas correlations with age showed correlation coefficient (r) values of 0.1377, 0.4185, 0.0597, -0.0993, and -0.0073, respectively. The r values between ACE2 protein and ACE2 mRNA with DPP-4 activity were -0.0668 and 0.1935, respectively. The values between both DPP-4 activity and ACE2 protein with MMP-2 activity were -0.0506 and 0.0898, respectively. Results reflected (1) MMP-2 activity is higher among males. (2) DPP-4 activity positively correlates with age. (3) Gender differences in DPP-4 activity, ACE2 protein, and mRNA levels were not statistically significant. (4) Age differences in MMP-2 activity, ACE2 expression were not significant. (5) Inter-enzyme correlations were not observed.
CONCLUSIONS: The differential activity of both MMP-2 and DPP-4 highlights their potential as risk factors for susceptibility to SARS-CoV-2 infection and COVID-19 severity in males and older groups.
AIMS: To study expression and activity profiles of implicated enzymes in SARS-CoV-2 infection in various age groups of both genders and discuss the possible impact of this on COVID-19 outcome.
METHODS: Saliva from different age groups was used to profile MMP-2 and gelatinase enzymes by zymography. Enzyme activities resulting from the cleavage of the H-Arg-Pro-pNA substrate were measured using a microplate reader. ACE2 transcription and translation were quantified using qRT-PCR and ELISA. Statistical analysis was used to identify the presence of significant differences.
RESULTS: Zymography demonstrated different gelatinase activities in a range of ∼180-30 kDa and showed distinct bands at ∼96, 72, 66, 60, and 40 kDa. Comparing MMP-2, DPP-4, DPP-4/ACE2 activities, ACE2 protein, and mRNA levels between genders showed p-values of 0.0142, 0.1056, 0.1859, 0.6411, and 0.4797, whereas correlations with age showed correlation coefficient (r) values of 0.1377, 0.4185, 0.0597, -0.0993, and -0.0073, respectively. The r values between ACE2 protein and ACE2 mRNA with DPP-4 activity were -0.0668 and 0.1935, respectively. The values between both DPP-4 activity and ACE2 protein with MMP-2 activity were -0.0506 and 0.0898, respectively. Results reflected (1) MMP-2 activity is higher among males. (2) DPP-4 activity positively correlates with age. (3) Gender differences in DPP-4 activity, ACE2 protein, and mRNA levels were not statistically significant. (4) Age differences in MMP-2 activity, ACE2 expression were not significant. (5) Inter-enzyme correlations were not observed.
CONCLUSIONS: The differential activity of both MMP-2 and DPP-4 highlights their potential as risk factors for susceptibility to SARS-CoV-2 infection and COVID-19 severity in males and older groups.