Tetrandrine suppresses activation of human subconjunctival fibroblasts in vitro.
OBJECTIVE
To investigate the effect of tetrandrine on activation of human subconjunctival fibroblasts (SCFs) in vitro.


METHODS
Effects of tetrandrine on proliferation, matrix expression, myofibroblast generation, and cell migration of SCF were examined in SCF cultures. Tetrandrine effect on TGFbeta 1/Smad2 signal and Smad7 expression was examined. Migration was evaluated by in vitro defect closure in a monolayer cell culture. Western blotting and immunocytochemistry were used.


RESULTS
Tetrandrine suppressed wound-induced cell migration (defect closure) and myofibroblast generation, but not cell proliferation in SCF cultures. It also decreased expression of fibronectin, collagen I, and alpha SMA. Adding tetrandrine increased protein level of Smad7 and suppressed Smad2 signal.


CONCLUSIONS
Tetrandrine suppresses Smad2 signal and fibrogenic responses in SCFs in association with Smad7 up-regulation, suggesting its potential therapeutic value to prevent excess scarring/fibrosis in conjunctiva following trabeculectomy or in patients with severe conjunctival inflammation.
Kitano
Ai
A
Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan. aikitano@wakayamamed.ac.jp
Yamanaka
Osamu
O
Ikeda
Kazuo
K
Ishida-Nishikawa
Iku
I
Okada
Yuka
Y
Shirai
Kumi
K
Saika
Shizuya
S
D000199:Actins / Q000378:metabolism; D044182:Benzylisoquinolines / Q000494:pharmacology*; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism; D003228:Conjunctiva / Q000166:cytology*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D051899:Smad2 Protein / Q000378:metabolism; D051906:Smad7 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
D000199:Actins; D044182:Benzylisoquinolines; D024042:Collagen Type I; D024061:Collagen Type III; D004365:Drugs, Chinese Herbal; D005353:Fibronectins; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; C497782:SMAD2 protein, human; C497806:SMAD7 protein, human; D051899:Smad2 Protein; D051906:Smad7 Protein; D053773:Transforming Growth Factor beta1; C009438:tetrandrine