Zhonghua yi xue za zhi · 2009
[Function of STAT1 in proliferation and differentiation of human hypertrophic scar fibroblast induced by connective tissue growth factor].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Tao Ling, Liu Jian-Yi, Li Shi-Rong, Dai Xia, Li Zhe
Аннотация
OBJECTIVE: To validate whether STAT1 participated in the process of CTGF-induced proliferation and differentiation of human hypertrophic scar fibroblast (hHSF) on account of past research.
METHODS: To cultivate hHSF with 6 patients' hypertrophic scar specimens together. Electrophoretic mobility shift assay (EMSA) was used to verify binding ability between DNA and STAT1 with the stimulus of different concentration CTGF (0, 5, 7.5, 10, 15 ng/ml) at 45th min and the stimulus of 10 ng/ml CTGF at different phase point (0, 10, 20, 30, 45, 60, 90, and 120 min). We divided cells into CTGF group, STAT1 ASODN group. STAT1 ASODN + CTGF group. control group. And MTT was used to detect the proliferation of hHSF on days1, 2 and 3, and RT-PCR to detect alpha-smooth muscle actin mRNA to follow the differentiation.
RESULTS: EMSA showed that the binding ability between STAT1 and DNA depended on the concentration of CTGF and peaked with the stimulation of 10ng/ml CTGF. And at the same time, it peaked at 45 - 60 min with 10 ng/ml CTGF. MTT showed that cell proliferation of CTGF group was much higher than that of control group (all P < 0.05). And those of STAT1 ASODN group and STAT1 ASODN + CTGF group were much lower than those of control group and CTGF group (all P < 0.05). RT-PCR showed that differentiation activation from fibroblast to myofibroblast of CTGF group, STAT1 ASODN group, STAT1 ASODN + CTGF group and control group were 0.78 +/- 0.08, 0.38 +/- 0.09, 0.76 +/- 0.10, and 0.40 +/- 0.12, respectively. Differentiation activation of STAT1 ASODN group and control group were much lower than those of CTGF group and STAT1 ASODN + CTGF group (all P < 0.05).
CONCLUSION: STAT1 ASODN is important in the process of proliferation of hHSF and it blocks the stimulation of CTGF on hHSF proliferation. The above result revealed that STAT1 participates in the process of hHSF proliferation induced by connective tissue growth factor.
METHODS: To cultivate hHSF with 6 patients' hypertrophic scar specimens together. Electrophoretic mobility shift assay (EMSA) was used to verify binding ability between DNA and STAT1 with the stimulus of different concentration CTGF (0, 5, 7.5, 10, 15 ng/ml) at 45th min and the stimulus of 10 ng/ml CTGF at different phase point (0, 10, 20, 30, 45, 60, 90, and 120 min). We divided cells into CTGF group, STAT1 ASODN group. STAT1 ASODN + CTGF group. control group. And MTT was used to detect the proliferation of hHSF on days1, 2 and 3, and RT-PCR to detect alpha-smooth muscle actin mRNA to follow the differentiation.
RESULTS: EMSA showed that the binding ability between STAT1 and DNA depended on the concentration of CTGF and peaked with the stimulation of 10ng/ml CTGF. And at the same time, it peaked at 45 - 60 min with 10 ng/ml CTGF. MTT showed that cell proliferation of CTGF group was much higher than that of control group (all P < 0.05). And those of STAT1 ASODN group and STAT1 ASODN + CTGF group were much lower than those of control group and CTGF group (all P < 0.05). RT-PCR showed that differentiation activation from fibroblast to myofibroblast of CTGF group, STAT1 ASODN group, STAT1 ASODN + CTGF group and control group were 0.78 +/- 0.08, 0.38 +/- 0.09, 0.76 +/- 0.10, and 0.40 +/- 0.12, respectively. Differentiation activation of STAT1 ASODN group and control group were much lower than those of CTGF group and STAT1 ASODN + CTGF group (all P < 0.05).
CONCLUSION: STAT1 ASODN is important in the process of proliferation of hHSF and it blocks the stimulation of CTGF on hHSF proliferation. The above result revealed that STAT1 participates in the process of hHSF proliferation induced by connective tissue growth factor.