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Change of UGBP in Lung Fibroblasts and Human Bronchial Epithelial and Primary Mechanism

Author: CaoShengTian
Tutor: LuoZiQiang
School: Central South University
Course: Physiology
Keywords: pulmonary fibrosis BLM TGF-β1 UGBP SIS3
CLC: R563
Type: Master's thesis
Year: 2011
Downloads: 10
Quote: 0
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Abstract

uteroglobin (UG) is a highly conserved, multi-functional small molecular secreting protein, secreted by the Clara cells. Its biological effects may depend on mediation of the uterus globin binding protein (Uteroglobin-binding protein, UGBP). But now, UGBP are still poorly understood at home and abroad and its research lags far behind its natural ligand of UG. In this study, we first observed the expression of UGBP on the mice model of pulmonary fibrosis at the overall level.Then, we examined the expression of UGBP in NIH3T3 and HBE induced by TGF-β1and explored its intracellular signal transduction to provide new clues for further exploration of mechanisim of UGBP expression in the process of pulmonary fibrosis.Methods:①The model of pulmonary fibrosis in mice was made by intratracheal delivery of bleomycin to mice and virify it by HE and Masson staining in morphology and by real-time PCR at the gene level. Then UGBP expression was detected by real-time PCR and tissue Microarray immunohistochemical.②UGBP mRNA expression in NIH3T3 and HBE cells was detected by real-time PCR;③Real-timePCR assay on UGBP mRNA Expression in NIH3T3 and HBE cells after application of smad3 protein inhibitor (SIS3). Results:1. HE staining showed:Fibroblast increased obviously after intratracheal delivery of bleomycin for 21th day and fibroblast foci was formed. Massson staining displayed:Collagen in section 7 Days showed almost no difference compared to control group, to14,21daysincreased significantly compared to control group. Real-timePCR results:mRNA expressions of TGF-beta increased significantly compared to control group on 7,14,21 days (p< 0.05), mRNA expression of typeⅢcollagen increased slightly in the first seven days,increased significantly on 14,21 days (p< 0.05).2. In the whole lung fibrosis model, real-timePCR results showed UGBP mRNA expression increased on 7 days compared to control group (p <0.05); to 14 days, UGBP mRNA expression increased,but no significant compared to control group(p>0.05); to 21 days, UGBP mRNA expression increased,but no significant compared to control group(p>0.05). Tissue Microarray immunohistochemical results showed that:UGBP protein has strong positive expression in bronchial epithelial cells, inflammatory cells and alveolare pithelialcells,selectivitystrong positive expression in fibroblast.UGBP protein expression in bronchial epithelial cells has stronger positive than alveolare pithelialcells and fibroblast. 3. BLM(1μg/ml) applied to NIH3T3 cells for 8h and 12h, the real-timePCR results suggested that BLM could decrease UGBP mRNA expression (8h,compared with control group p<0.05; 12h, compared with control group, p<0.01)4. TGF-β1(10ng/ml) applied to NIH3T3 cells for 8h and 12h, the real-timePCR results suggested that TGF-β1 could down-regulate UGBP mRNA expression(8h,compared with control group p<0.05; 12h, compared with control group, p<0.01); compared with serum-free group, serum could down-regulate UGBP mRNA expression (p<0.01).5. TGF-β1(lOng/ml) in HBE cells for 12h, real-timePCR results: TGF-β1 could upregulate UGBP mRNA expression (p<0.01)6. SIS3 (3μmol/L) can reduce the TGF-β1 on UGBP mRNA down-regulation expression in NIH3T3,compared with TGF-β1 (10ng/ml) group (p<0.01); SIS3 (3μmol/L) also can reduce the effect of TGF-β1 on UGBP mRNA up-regulation expression in HBE,compared with TGF-β1 (10ng/ml) group (p<0.05)Conclusion:1. In lung fibrosis, UGBP mRNA expression is a transient increase. 2.TGF-β1 has different effect on UGBP mRNA expression in NIH3T3 and HBE in vitro conditions.It inhibits and promot UGBP mRNA expression in NIH3T3 and HBE respectively.3. Regulation of UGBP mRNA expression in NIH3T3 and HBE cells by TGF-β1 was mediated by the smad signaling pathway.

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CLC: > Medicine, health > Internal Medicine > Respiratory system and chest diseases > Pulmonary disease
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