Objective To determine the effects of insulin-like growth factor-1 （IGF-1） on the expression of preprotachykinin （PPT） mRNA encoding substance P （SP） and calcitonin gene-related peptide （CGRP） mRNA in cultured...Objective To determine the effects of insulin-like growth factor-1 （IGF-1） on the expression of preprotachykinin （PPT） mRNA encoding substance P （SP） and calcitonin gene-related peptide （CGRP） mRNA in cultured dorsal root ganglion （DRG） neurons with excitotoxicity induced by glutamate （Glu）. Methods DRGs were dissected from embryonic day 15 Wistar rats. DRG neurons were dissociated and cultured for 48 h and then exposed to Glu （0.2 mmol/L） or Glu （0.2 mmol/L） plus IGF- 1 （5 nmol/L, 10 nmol/L and 20 nmol/L） for 12 h. The DRG neurons in control group were exposed to only growth media throughout the experiment. After that, the living DRG neurons were observed under inverted phase contrast microscope and microphotographs were taken. The expression levels of PPT and CGRP mRNAs were detected by reverse transcriptionpolymerase chain reaction （RT-PCR）. Results IGF-1 could inhibit Glu-induced shortening of neurite. Besides, IGF-1 could significantly increase the levels ofPPT mRNA and CGRP mRNA in primary cultured DRG neurons with Glu-induced excitotoxicity, in a dose-dependent manner. Conclusion IGF-1 may exert neuroprotective effects on DRG neurons against Glu-induced excitotoxicity, probably through regulating the expression levels of PPT and CGRP mRNAs.展开更多
Objective: To investigate the effect of substance P (SP) on gene expression of transforming growth factor β-1 (TGFβ-1), transforming growth factor receptor-1 (TGFR-1) and transforming growth factor receptor-2 (TGFR-...Objective: To investigate the effect of substance P (SP) on gene expression of transforming growth factor β-1 (TGFβ-1), transforming growth factor receptor-1 (TGFR-1) and transforming growth factor receptor-2 (TGFR-2) in fibroblasts cultured in vitro from rat’s granulation tissues. Methods: The fibroblasts from the granulation tissues in the skeletal muscle of rat’s hind limbs injured by formaldehyde were cultured in vitro. When different concentrations (10 -9-10 -5 mol/L) of SP were added into the culture medium, the changes of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the cultured fibroblasts were observed with reverse transcription polymerase chain reaction at different intervals (0, 3, 6, 12 and 24 hours after incubation). Results: The gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts cultured from rat’s granulation tissues was up-regulated by SP. The peak level of the mRNA expression was found at 10 -8 mol/L SP and the up-regulation effect was not found at 10 -5 mol/L and 10 -6 mol/L. The peak levels of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts treated with SP were achieved at 6 and 12 hours, respectively. Conclusions: SP has up-regulation effect on the gene expression of TGFβ-1, TGFR-1 and TGFR-2 in fibroblasts from rat’s granulation tissues in vitro, and the effect is related to different stimulating concentrations of SP. It may be concerned with proliferation and differentiation of fibroblasts and formation of scar tissues during wound healing.展开更多
基金supported by the Natural Sciences Foundation of Shandong Province, China(No. Z2006C06)the Science and Technology Development Project of Jinan Municipality of Shandong Province,China (No. 200705083, 200807046)
文摘Objective To determine the effects of insulin-like growth factor-1 （IGF-1） on the expression of preprotachykinin （PPT） mRNA encoding substance P （SP） and calcitonin gene-related peptide （CGRP） mRNA in cultured dorsal root ganglion （DRG） neurons with excitotoxicity induced by glutamate （Glu）. Methods DRGs were dissected from embryonic day 15 Wistar rats. DRG neurons were dissociated and cultured for 48 h and then exposed to Glu （0.2 mmol/L） or Glu （0.2 mmol/L） plus IGF- 1 （5 nmol/L, 10 nmol/L and 20 nmol/L） for 12 h. The DRG neurons in control group were exposed to only growth media throughout the experiment. After that, the living DRG neurons were observed under inverted phase contrast microscope and microphotographs were taken. The expression levels of PPT and CGRP mRNAs were detected by reverse transcriptionpolymerase chain reaction （RT-PCR）. Results IGF-1 could inhibit Glu-induced shortening of neurite. Besides, IGF-1 could significantly increase the levels ofPPT mRNA and CGRP mRNA in primary cultured DRG neurons with Glu-induced excitotoxicity, in a dose-dependent manner. Conclusion IGF-1 may exert neuroprotective effects on DRG neurons against Glu-induced excitotoxicity, probably through regulating the expression levels of PPT and CGRP mRNAs.
基金ThisworkwassupportedbyMajorStateBasicResearchDevelopmentProgramofChina (No .G19990 5 4 2 0 4 )
文摘Objective: To investigate the effect of substance P (SP) on gene expression of transforming growth factor β-1 (TGFβ-1), transforming growth factor receptor-1 (TGFR-1) and transforming growth factor receptor-2 (TGFR-2) in fibroblasts cultured in vitro from rat’s granulation tissues. Methods: The fibroblasts from the granulation tissues in the skeletal muscle of rat’s hind limbs injured by formaldehyde were cultured in vitro. When different concentrations (10 -9-10 -5 mol/L) of SP were added into the culture medium, the changes of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the cultured fibroblasts were observed with reverse transcription polymerase chain reaction at different intervals (0, 3, 6, 12 and 24 hours after incubation). Results: The gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts cultured from rat’s granulation tissues was up-regulated by SP. The peak level of the mRNA expression was found at 10 -8 mol/L SP and the up-regulation effect was not found at 10 -5 mol/L and 10 -6 mol/L. The peak levels of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts treated with SP were achieved at 6 and 12 hours, respectively. Conclusions: SP has up-regulation effect on the gene expression of TGFβ-1, TGFR-1 and TGFR-2 in fibroblasts from rat’s granulation tissues in vitro, and the effect is related to different stimulating concentrations of SP. It may be concerned with proliferation and differentiation of fibroblasts and formation of scar tissues during wound healing.
