hrpe arpe 19 cells  (ATCC)

Journal: Experimental and therapeutic medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling.

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Figure 1. Complement C3a and C5a promote cell viability and inflammation of HRPE cells. (A) Complement C3a and (B) C5a protein concentrations in patients with RRDCD (n=20) and idiopathic epimacular membrane as a control (n=20). HRPE cells were treated with different concentrations of recombinant human complement component C3a or C5a, and the cell viability following (C) C3a or (D) C5a treatment, and the release of (E) TNF‑α, (F) IL‑1β, (G) IL‑6, (H) PGE2 and (I) IL‑10 were measured. *P<0.05, **P<0.01, ***P<0.001 compared with control or 0 µg/ml. OD450, optical density at 450 nm; PGE2, prosta‑ glandin E2; RRDCD, rhegmatogenous retinal detachment associated with choroidal detachment; HRPE, human retinal pigment epithelium.

Article Snippet: Human RPE (HRPE) cells (ARPE‐19; CRL‐2302) purchased from the American Type Culture Collection were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a humidified incubator with 5% CO2 at 37 ̊C.

Techniques: Membrane, Control, Recombinant

Journal: Experimental and therapeutic medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling.

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Figure 2. Complement C3a and C5a enhance C3aR and C5aR expression and activate NF‑κB signaling. The mRNA expression levels of (A) C3aR and (B) C5aR in HRPE cells treated with different doses of recombinant human complement component C3a or C5a. (C) The protein expression levels of C3aR, phosphorylation of NF‑κB (p‑NF‑κB), and expression of NF‑κB in HRPE cells treated with different doses of recombinant human complement component C3a. (D) Protein expression levels of C5aR, phosphorylation of NF‑κB and expression of NF‑κB in HRPE cells treated with different doses of recombinant human complement component C5a. *P<0.05, **P<0.01, ***P<0.001 relative to 0 µg/ml. p‑NF‑κB, phosphorylated NF‑κB; C5aR, C5a receptor; HRPE, human retinal pigment epithelium.

Article Snippet: Human RPE (HRPE) cells (ARPE‐19; CRL‐2302) purchased from the American Type Culture Collection were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a humidified incubator with 5% CO2 at 37 ̊C.

Techniques: Expressing, Recombinant, Phospho-proteomics

Journal: Experimental and therapeutic medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling.

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Figure 3. Complement C3a and C5a aggravate inflammation in HRPE cells via the NF‑κB signaling pathway. HRPE cells were treated with recombinant human complement component C3a (2 µg/ml) or C5a (1 µg/ml) with or without NF‑κB inhibitor PDTC (10 µM), and the release of (A) TNF‑α, (B) IL‑1β, (C) IL‑6, (D) PGE2 and (E) IL‑10 was measured by ELISA. ***P<0.001 relative to control; ###P<0.001 relative to C3a or C5a treatment. PGE2, prostaglandin E2; HRPE, human retinal pigment epithelium.

Article Snippet: Human RPE (HRPE) cells (ARPE‐19; CRL‐2302) purchased from the American Type Culture Collection were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a humidified incubator with 5% CO2 at 37 ̊C.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Control

Journal: Experimental and therapeutic medicine

Article Title: The complement C3a-C3aR and C5a-C5aR pathways promote viability and inflammation of human retinal pigment epithelium cells by targeting NF-κB signaling.

doi: 10.3892/etm.2022.11420

Figure Lengend Snippet: Figure 4. Complement C3aR and C5aR antagonist inhibit inflammation and NF‑κB signaling in HRPE cells challenged with complement C3a and C5a. HRPE cells were treated with recombinant human complement component C3a (2 µg/ml) and with or without C3aR antagonist SB290157 (20 µM), and the release of (A) TNF‑α, (B) IL‑1β, (C) IL‑6, (D) PGE2 and (E) IL‑10 was determined by ELISA. HRPE cells were treated with recombinant human complement component C5a (1 µg/ml) and with or without C5aR antagonist CCX168 (2 µM), and the release of (F) TNF‑α, (G) IL‑1β, (H) IL‑6, (I) PGE2 and (J) IL‑10 was determined by ELISA. (K) The phosphorylation of NF‑κB and expression of NF‑κB in HRPE cells treated with recombinant human complement component C3a (2 µg/ml) and with or without C3aR antagonist SB290157 (20 µM) were determined by western blot. (L) The phosphorylation of NF‑κB and expression of NF‑κB in HRPE cells treated with recombinant human complement component C5a (1 µg/ml) and with or without C5aR antagonist CCX168 (2 µM) were determined by western blot. ***P<0.001 relative to control; ###P<0.001 relative to C3a or C5a treatment. p‑NF‑κB, phosphorylated NF‑κB; C5aR, C5a receptor; HRPE, human retinal pigment epithelium; CCX, CCX168; SB, SB290157.

Article Snippet: Human RPE (HRPE) cells (ARPE‐19; CRL‐2302) purchased from the American Type Culture Collection were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a humidified incubator with 5% CO2 at 37 ̊C.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Expressing, Western Blot, Control