Implication of the PI3K/Akt/mTOR Pathway in the Process of Incompetent Valves in Patients with Chronic Venous Insufficiency and the Relationship with Aging

Abstract

Chronic venous insufficiency (CVI) is a multifactorial disease, commonly caused by valvular incompetence (clinically diagnosed by venous reflux) and venous hypertension. The incidence of these factors clearly increases with patient age, and aging is one of the risk factors involved. The activity of the PI3K/Akt/mTOR pathway is considered fundamental in vascular pathologies, and understanding its involvement would help in the development of possible therapeutic targets. This is an observational, analytical, and prospective cohort study that reviewed 110 patients with CVI scheduled to undergo stratified saphenectomy. They were distributed according to the presence (R = 81) or absence (NR = 29) of valvular incompetence (venous reflux) diagnosed clinically. Each of the groups was further divided according to age, with a cutoff point of 50 years (NR < 50 = 13, NR ≥ 50 = 16, R < 50 = 32, and R ≥ 50 = 49). The involvement of the PI3K/Akt/mTOR pathway, as well as that of HIF-1α and HIF-2α and of CD4+, CD8+, and CD19+ cells and mastocytes, was assessed. Saphenous vein tissue samples obtained during surgery were processed for RT-qPCR and immunohistochemistry. Patients with venous reflux showed a significant increase in mRNA and protein expression levels for PI3K/mTOR and HIF-1α/HIF-2α. The number of mast cells was significantly elevated in the R group. In distribution by age, PI3K/Akt/mTOR and HIF-1α were significantly higher in R < 50 patients. Furthermore, these patients had a significant increase in the number of CD4+, CD8+, and CD19+ cells and mastocytes in the saphenous vein wall. These findings provide a basis for the possible existence of changes in PI3K/Akt/mTOR pathway expression in young patients, with potential accelerated asynchronous aging that is enhanced by CVI.

Pregnancy‐associated venous insufficiency course with placental and systemic oxidative stress

“…De novo primers or primers that were specific for all genes of interest were designed (Table S1) using the online applications Primer-BLAST 39 and AutoDimer. 40 RNA was extracted using the guanidine-phenol-chloroform isothiocyanate method described by Ortega et al 41 The qPCR was performed in a StepOnePlus™ System (Applied Biosystems-Life Technologies) using the relative standard curve method. The reaction was performed as follows: 5 µl of each sample containing cDNA diluted 1/20 in nuclease-free water was mixed with 10 µL of iQ™ SYBR ® Green Supermix (Bio-Rad Laboratories), 1 µL of forward primer (6 μmol/L), 1 µL of reverse primer (6 μmol/L) and 3 µL of DNase-and RNase-free water in a MicroAmp ® 96-well plate (Applied Biosystems-Life Technologies) for a total reaction volume of 20 µL.…”

“…The complete study was performed with 114 patients, including 62 women with VI during pregnancy and 52 control women without evidence of VI during pregnancy. The mean age is 33 [22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40] VI and 34 [27][28][29][30][31][32][33][34][35][36][37][38][39][40][41] HC. Not exist significant differences in the mean age of patients (P = .06915).…”

“…Not exist significant differences in the mean age of patients (P = .06915). The gestational age (weeks) is 40.5 [39-41.5] VI and 41 [39][40][41][42] HC. The number of caesarean delivery is 12 (19.4%) in VI and 9 (17.3%) in HC.…”

Increased Angiogenesis and Lymphangiogenesis in the Placental Villi of Women with Chronic Venous Disease during Pregnancy

“…Sixty-two were clinically diagnosed with CVD, with a median age of 33 years (22-40) and median gestational age (weeks) of 40.5 (39-41.5). In parallel, 52 controls with no history of CVD (HC) were studied, with a median age of 34 years (27-41) and median gestational age (weeks) of 41 (39)(40)(41)(42). This study was conducted according to the basic ethical principles of autonomy, beneficence, non-maleficence, and distributive justice.…”

“…RNA extraction was executed according to the guanidinium thiocyanate-phenol-chloroform method described by Ortega et al [41]. RT-qPCR was performed in a StepOnePlus™ System (Applied Biosystems -Life Technologies, Waltham, MA, USA), with the standard curve method.…”