Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 in vitro

2020 ◽  
Vol 1 (1) ◽  
pp. 17-23
Author(s):  
Hua Zhuang ◽  
◽  
Jing Wu ◽  
Wei Xu ◽  
Jian-Zhang Hu ◽  
...  
Keyword(s):  
Smooth Muscle Actin ◽  
Negative Control ◽  
Tissue Growth ◽  
Lens Epithelium ◽  
Cell Counting ◽  
Human Lens ◽  
Posterior Capsular Opacification ◽  
Epithelium Cell ◽  
Human Lens Epithelium

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-smooth muscle actin (α-SMA) in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated small interfering RNA (siRNA) targeting CTGF. METHODS: HLECs were transfected with siRNA targeting CTGF, labeled with 5’-fluorescein isothiocyanate (5’-FITC) and coated with lipofectamine. The transfection ratio was evaluated via fluorescence intensity. Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs. Quantitative reverse transcription-polymerase chain reaction (RT-PCR), cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection. RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine. The transfection ratio reached 95% at 24h. The proliferation of HLECs was inhibited by siRNA after 72h transfection. The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h. This effect was not found in negative control siRNA. CONCLUSION: siRNA targeting CTGF decrease CTGF and α-SMA expression in HLECs, which is a potential therapeutic strategy for posterior capsular opacification.

scholarly journals Somatic Variants in the Human Lens Epithelium: A Preliminary Assessment

2016 ◽  
Vol 57 (10) ◽  
pp. 4063 ◽  
Author(s):  
Rosana Mesa ◽  
Manoj Tyagi ◽  
George Harocopos ◽  
David Vollman ◽  
Steven Bassnett
Keyword(s):  
Lens Epithelium ◽  
Human Lens ◽  
Preliminary Assessment ◽  
Human Lens Epithelium

An Immunoperoxidase Study of Human Lens Epithelium

1990 ◽  
Vol 2 (1) ◽  
pp. 71-73 ◽  
Author(s):  
Margaret J. Jeffrey ◽  
W.T. Green ◽  
D.L. Boase ◽  
M.N. Jeffrey
Keyword(s):  
Lens Epithelium ◽  
Human Lens ◽  
Human Lens Epithelium

Store-operated Calcium Entry-associated Regulatory Factor Regulates Airway Inflammation and Airway Remodeling in Asthma Mice Models

2021 ◽  
Author(s):  
Yi-min Wang ◽  
Wen-juan Xu ◽  
Lin-li Xiang ◽  
Mei Ding ◽  
Jin-jin Zhang ◽  
...  
Keyword(s):  
Airway Inflammation ◽  
Airway Remodeling ◽  
Protective Role ◽  
Negative Control ◽  
Calcium Entry ◽  
Cell Counting ◽  
Airway Smooth Muscle Cells ◽  
Regulatory Factor ◽  
Store Operated Calcium Entry

Background: Store-operated calcium entry (SOCE) is involved in the pathogenesis of airway inflammation and remodeling in asthma. Store-operated calcium entry-associated regulatory factor (SARAF) can down-regulate SOCE. Objective: We sought to investigate the role of SARAF in the regulation of airway inflammation and remodeling in asthma mice models, as well as in the functional regulation of human airway smooth muscle cells (hASMCs). Methods: Balb/c mice were sensitized and challenged with ovalbumin to establish the asthma mice models. Mice were transfected with lentivirus, which expressed the SARAF gene + GFP or the negative control gene + GFP. Airway resistance was measured with the animal pulmonary function system. Airway inflammation and remodeling were evaluated via histological staining. In vitro cultured hASMCs were transfected with scrambled small interfering RNA(siRNA) or SARAF-specific siRNA respecitvely. The proliferation, migration rate, hypertrophy and SOCE activity of hASMCs were examined with cell counting kit 8, wound healing test, bright field imaging and Ca2+ fluorescence imaging, respectively. SARAF expression was measured by quantitative real-time-PCR. Results: Asthma mice models showed decreased SARAF mRNA expression in the lungs. SARAF overexpression attenuated airway inflammation, resistance and also remodeling. Downregulation of SARAF expression with siRNA promoted the proliferation, migration, hypertrophy and SOCE activity in hASMCs. Conclusions: SARAF plays a protective role against airway inflammation and remodeling in asthma mice models by blunting SOCE; SARAF may also be a functional regulating factor of hASMCs.

scholarly journals Ethylene glycol coated nanoceria protects against oxidative stress in human lens epithelium

RSC Advances ◽  
2019 ◽  
Vol 9 (29) ◽  
pp. 16596-16605 ◽  
Author(s):  
Belal I. Hanafy ◽  
Gareth W. V. Cave ◽  
Yvonne Barnett ◽  
Barbara Pierscionek
Keyword(s):  
Oxidative Stress ◽  
Ethylene Glycol ◽  
Lens Epithelium ◽  
Human Lens ◽  
Human Lens Epithelium

Nanoceria (<5 nm), stabilised with ethylene glycol, protects human lens epithelium from oxidative stress and exhibits multicoloured photoluminescence.

Sign in / Sign up

Export Citation Format

Share Document