Abdellatif Mentalla Motaz, El Amir Azza, Shalaby Said, Kirschfink Michael, Nadeem Rola, Abd-Elshafy Dina Nadeem, Bahgat Mahmoud Mohamed
Department of Therapeutic Chemistry, Institute of Pharmaceutical and Drug Industries Research, the National Research Centre, Dokki, Giza 12311, Egypt; Research Group Immune- and Biomarkers for Infection, the Centre of Excellence for Advanced Sciences, the National Research Centre, Giza 12311, Egypt.
Zoology Department, Faculty of Science, Cairo University, Giza 12613, Egypt.
J Genet Eng Biotechnol. 2025 Sep;23(3):100509. doi: 10.1016/j.jgeb.2025.100509. Epub 2025 Jun 17.
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.
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.
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.
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.
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.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）通过血管紧张素转换酶2（ACE2）进入人体细胞。计算机模拟实验还表明，SARS-CoV-2在细胞进入过程中可能与二肽基肽酶4（DPP-4）结合。此外，基质金属蛋白酶2（MMP-2）参与SARS-CoV-2细胞融合过程。
研究SARS-CoV-2感染中涉及的酶在不同年龄组和性别的表达及活性谱，并讨论其对2019冠状病毒病（COVID-19）结局的可能影响。
使用来自不同年龄组的唾液，通过酶谱法分析MMP-2和明胶酶。使用酶标仪测量H-Arg-Pro-pNA底物裂解产生的酶活性。使用定量逆转录聚合酶链反应（qRT-PCR）和酶联免疫吸附测定（ELISA）对ACE2的转录和翻译进行定量。采用统计分析确定是否存在显著差异。
酶谱法显示在约180 - 30 kDa范围内存在不同的明胶酶活性，并在约96、72、66、60和40 kDa处显示出明显条带。比较不同性别之间的MMP-2、DPP-4、DPP-4/ACE2活性、ACE2蛋白和mRNA水平，p值分别为0.0142、0.1056、0.1859、0.6411和0.4797，而与年龄的相关性显示相关系数（r）值分别为0.1377、0.4185、0.0597、-0.0993和-0.0073。ACE2蛋白和ACE2 mRNA与DPP-4活性之间的r值分别为-0.0668和0.1935。DPP-4活性和ACE2蛋白与MMP-2活性之间的值分别为-0.0506和0.0898。结果表明：（1）男性的MMP-2活性较高。（2）DPP-4活性与年龄呈正相关。（3）DPP-4活性、ACE2蛋白和mRNA水平的性别差异无统计学意义。（4）MMP-2活性、ACE2表达的年龄差异不显著。（5）未观察到酶之间的相关性。
MMP-2和DPP-4的差异活性突出了它们作为男性和老年群体中SARS-CoV-2感染易感性和COVID-19严重程度风险因素的潜力。
