title: [Effects of exogenous TGF-β3 on the expression of endogenous TGF-β3 in hepatic stellate cell-T6 (HSC-T6)].
abstract: OBJECTIVE
  To investigate the effects of exogenous TGF-β3 on the expression of endogenous TGF-b3 in hepatic stellate cell (HSC).
  
  
  METHODS
  HSCs were cultured and divided into two groups: TGF-β3 group and blank control group, the cells of TGF-β3 group were exposed to TGF-b3 (10 ng/ml), whereas the blank control group was not treated. The cells were incubated in the presence of exogenous TGF-β3 and then (1) were harvested at 0h, 1h, 2h, 4h, 12h, 24h, and real time PCR was performed to detect the mRNA expression of endogenous TGF-β3. (2) The cells were collected at 0h, 1h, 6h, 12h, and western-blot was used to detect the protein synthesis of endogenous TGF-β3 in HSC; (3) The cell culture supernatant was harvested at 0h, 1h, 2h, 4h, 8h, 14h, 24h, and ELISA was performed to measure the total protein of extracellular TGF-β3; HSCs were treated with TGF-β3 (10 ng/ml) for 2h. The cells were then incubated in serum-free medium and the cell culture supernatant was harvested at 2.25h, 2.5h, 3h, 4h, 6h, 10h and 14h. ELISA was used to detect the extracellular secret ion of endogenous TGF-β3 by HSCs.
  
  
  RESULTS
  (1) Exogenous TGF-β3 treatment induced a marked increase in TGF-β3 mRNA expression. By 2h of exogenous TGF-β3 treatment, maximal TGF-β3 mRNA expression levels (2.796 ± 0.518) of 2.74 fold above control values (1.022 ± 0.038) was reached (P < 0.05). Thereafter, TGF-β3 mRNA expression level declined, and the expression level was maintained at level of 1.45-fold for at least 10h and was 1.18-fold above control values by 24h TGF-β3 treatment (P < 0.05); (2) No significant difference about the intracellular protein expression level of endogenous TGF-β3 was found between two groups. (P > 0.05); (3) The total expression level of TGF-β3 reached a peak [(18.931 ± 2.904) ng/ml] at 4h after TGF-β3 treatment (1.89-fold higher than basic TGF-β3 (10 ng/ml). After that, it slowly declined. The expression peak [(0.835 ± 0.027) ng/ml] induction of extracellular secreted TGF-β3 was at 3h (32.12-fold higher than control [(0.026 ± 0.022) ng/ml], (P < 0.05). Thereafter, TGF-β3 slowly decreased after the peak time, and their expressions were still statistically significant as compared to the control (P < 0.05).
  
  
  CONCLUSIONS
  Exogenous TGF-β3 could increase the expression of endogenous TGF-β3 mRNA and extracellular secreted TGF-β3 protein obviously.
authors.affiliation: Department of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Liang
authors.initials: L
authors.lastname: Deng
authors.forename: Wei
authors.initials: W
authors.lastname: Qian
authors.forename: Jian-ning
authors.initials: JN
authors.lastname: Zhou
authors.forename: Ke-shu
authors.initials: KS
authors.lastname: Xu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism*; D051381:Rats; D053782:Transforming Growth Factor beta3 / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D053782:Transforming Growth Factor beta3