Fig 2: Plasma Obese-EVs induce renal tubule injury and RAS activation in PTECs via transport of miR-6869-5p. (A–D) After in vitro transfection of miR-6869-5p mimic and miR-NC-mimic, the protein levels of AGT, ACE, ACE2, AT1 (A,B), KIM-I, and NAGAL (C,D) were analyzed by Western blotting (n = 3). (E,F) After transfection with miR-6869-5p inhibitor or miR-NC-inhibitor for 48 h, PTECs were treated with PBS, Lean-EVs, or Obese-EVs. The protein levels of AGT, ACE, ACE2, AT1 (E,F), KIM-I, and NGAL (G,H) were analyzed by Western blotting (n = 3). Data are presented as mean ± SD; **P <0.001 vs. PBS, *P <0.001 vs. Lean-EVs. EVs, extracellular vesicles; PTECs, Proximal tubular epithelial cells; AGT, angiotensinogen; ACE, angiotensin-converting enzyme; ACE2, angiotensin-converting enzyme 2; AT1, angiotensin 1; KIM-1, kidney injury molecule-1; NGAL, neutrophil gelatinase-associated lipocalin.

Fig 3: Tissue renin–angiotensin systems in endometrial cancer. In tumour tissue, upregulation of cathepsin D and (P)RR allows for greater activation of prorenin, potentially leading to increased enzymatic cleavage of Ang I from AGT. Additionally, increased ACE1 abundance may lead to increased production of the biologically active Ang II. Together, upregulation of the pro-angiogenic/pro-proliferative components may lead to increased activation of both (P)RR- and AGTR1-mediated intracellular signalling cascades to stimulate the production of TGFB1 and PI3KR1 and promote tumourigenesis. The role of increased ACE2 and presence of MAS1 and AGTR2 may enhance tumour development by stimulating vascularisation; however, this is unknown.

Fig 4: Obesity leads to renal tubule injury and RAS activation in humans and mice. (A–C) Representative images (A), body weight curves (B), and bodyweight levels after 18 weeks (C), of obese and lean mice. (D,E) Representative immunohistochemical staining of AGT, AT1, KIM1, and NGAL in kidney sections of lean and obese mice (Original magnification 200×). AGT, angiotensinogen; AT1, angiotensin 1; KIM-1, kidney injury molecule-1; NGAL, neutrophil gelatinase-associated lipocalin. *P < 0.001; ***P < 0.05.

Fig 5: Effects of plasma EVs on renal tubule injury and RAS activation in obesity. (A–C) After treating PTECs with PBS(negative control), Lean-EVs, or Obese-EVs, the relative mRNA and protein levels of AGT, ACE, ACE2, and AT1 were analyzed by qRT-PCR (A) and Western blotting (B,C) (n = 3 per group). (D,E) Levels of KIM1 and NGAL protein in PTECs treated with PBS, Lean-EVs, or Obese-EVs. Data are presented as mean ± SD; **P < 0.001 vs. PBS, *P < 0.001 vs. Lean-EVs. EVs, extracellular vesicles; PTECs, Proximal tubular epithelial cells; KIM-1, kidney injury molecule-1; NGAL, neutrophil gelatinase-associated lipocalin; AGT, angiotensinogen; ACE, angiotensin-converting enzyme; ACE2, angiotensin-converting enzyme 2; AT1, angiotensin 1.