PMID- 38028520
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231201
IS  - 2222-3959 (Print)
IS  - 2227-4898 (Electronic)
IS  - 2222-3959 (Linking)
VI  - 16
IP  - 11
DP  - 2023
TI  - In vitro protective effect of recombinant prominin-1 combined with microRNA-29b 
      on N-methyl-D-aspartate-induced excitotoxicity in retinal ganglion cells.
PG  - 1746-1755
LID - 10.18240/ijo.2023.11.03 [doi]
AB  - AIM: To determine the in vitro protective effect of recombinant prominin-1 
      (Prominin-1)+microRNA-29b (P1M29) on N-methyl-D-aspartate (NMDA)-induced 
      excitotoxicity in retinal ganglion cells (RGCs). METHODS: RGC-5 cells were 
      cultured, and NMDA-induced excitotoxicity at the range of 100-800 micromol/L was 
      assessed using the MTT assay. NMDA (800 micromol/L) was selected as the appropriate 
      concentration for preparing the cell model. To evaluate the protective effect of 
      P1M29 on the cell model, Prominin-1 was added at the concentration of 1-6 ng/mL 
      for 48h, and the cell survival was investigated with/without microRNA-29b. After 
      obtaining the appropriate concentration and time of P1M29 at 48h, real-time 
      polymerase chain reaction (PCR) was utilized to detect the relative mRNA 
      expression of vascular endothelial growth factor (VEGF) and transforming growth 
      factor (TGF)-beta2. Western blot detection was applied to measure the 
      phosphorylation levels of protein kinase B (AKT) and extracellular regulated 
      protein kinases (ERK) in RGC-5 cells after treatment with Prominin-1. Apoptosis 
      study of the cell model was conducted by flow cytometry for estimating the 
      anti-apoptotic effect of P1M29. Immunofluorescence analysis was used to analyze 
      the expression levels of VEGF and TGF-beta2. RESULTS: MTT cytotoxicity assays 
      demonstrated that P1M29 group had significantly higher cell survival rate than 
      Prominin-1 group (P<0.05). Real-time PCR data indicated that the expression 
      levels of VEGF were significantly increased in both Prominin-1 and P1M29 groups 
      compared NMDA and microRNA-29b group (P<0.05), while TGF-beta2 were significantly 
      decreased in both microRNA-29b and P1M29 groups compared NMDA and Prominin-1 
      group (P<0.05). Western blot results showed that both Prominin-1 and P1M29 groups 
      significantly increased the phosphorylation levels of AKT and ERK compared to 
      NMDA and microRNA-29b groups (P<0.05). Flow cytometry analysis revealed that 
      P1M29 could prevent RGC-5 cell apoptosis in the early stage of apoptosis, while 
      immunofluorescence results showed that P1M29 group had higher expression of VEGF 
      and lower expression of TGF-beta2 with a stronger green fluorescence than NMDA 
      group. CONCLUSION: Prominin-1 combined with microRNA-29b can provide a suitable 
      therapeutic option for ameliorating NMDA-induced excitotoxicity in RGC-5 cells.
CI  - International Journal of Ophthalmology Press.
FAU - Li, Jun-Hua
AU  - Li JH
AD  - Eye Hospital and School of Ophthalmology and Optometry, Wenzhou Medical 
      University, Wenzhou 325027, Zhejiang Province, China.
AD  - Eye Hospital of Wenzhou Medical University Hangzhou Branch, Hangzhou 310020, 
      Zhejiang Province, China.
FAU - Yu, Guan-Shun
AU  - Yu GS
AD  - Eye Hospital and School of Ophthalmology and Optometry, Wenzhou Medical 
      University, Wenzhou 325027, Zhejiang Province, China.
AD  - Eye Hospital of Wenzhou Medical University Hangzhou Branch, Hangzhou 310020, 
      Zhejiang Province, China.
FAU - Wang, Yu-Da
AU  - Wang YD
AD  - Eye Hospital and School of Ophthalmology and Optometry, Wenzhou Medical 
      University, Wenzhou 325027, Zhejiang Province, China.
AD  - Eye Hospital of Wenzhou Medical University Hangzhou Branch, Hangzhou 310020, 
      Zhejiang Province, China.
FAU - Li, Tian-Kun
AU  - Li TK
AD  - Tianjin Eye Hospital, Tianjin 300020, China.
LA  - eng
PT  - Journal Article
DEP - 20231118
PL  - China
TA  - Int J Ophthalmol
JT  - International journal of ophthalmology
JID - 101553860
PMC - PMC10626362
OTO - NOTNLM
OT  - Prominin-1
OT  - microRNA-29b
OT  - transforming growth factor-beta2
OT  - vascular endothelial growth factor
EDAT- 2023/11/29 18:42
MHDA- 2023/11/29 18:43
PMCR- 2023/11/18
CRDT- 2023/11/29 17:14
PHST- 2022/11/18 00:00 [received]
PHST- 2023/08/15 00:00 [accepted]
PHST- 2023/11/29 18:43 [medline]
PHST- 2023/11/29 18:42 [pubmed]
PHST- 2023/11/29 17:14 [entrez]
PHST- 2023/11/18 00:00 [pmc-release]
AID - ijo-16-11-1746 [pii]
AID - 10.18240/ijo.2023.11.03 [doi]
PST - epublish
SO  - Int J Ophthalmol. 2023 Nov 18;16(11):1746-1755. doi: 10.18240/ijo.2023.11.03. 
      eCollection 2023.