Effects Of Cigarette Smoke Extract On Skin Fibroblast Damage And Collagen Synthesis In Vitro
Keywords
Cigarette Smoke Extract, Fibroblasts, Collagen, Skin
Abstract
Abstract: Objective To explore the damaging effect of cigarette smoke extract (CSE) on skin fibroblasts cultured in vitro, and to observe the effect of CSE on the synthesis of type ¢ñ collagen in fibroblasts. Methods The changes in cell morphology and structure after exposure to the poison were observed under a light microscope; the tetramethylazolium salt (MTT) colorimetric method was used to detect the effects of different concentrations of CSE on the viability of skin fibroblasts in vitro at different times. And draw the growth curve of cells at different concentrations; ¦Â-galactosidase staining method detects the changes of ¦Â-galactosidase in fibroblasts after 5 generations of CSE; flow cytometry detects the changes of ¦Â-galactosidase in fibroblasts under different concentrations of CSE. Cell cycle changes at different exposure times; RT-PCR detection of the effects of different concentrations of CSE on the expression of type I procollagen mRNA in fibroblasts. Results After exposure, the cells showed typical damaged cell morphology under the light microscope. As the concentration of CSE exposure increased and the exposure time prolonged, cell viability decreased significantly. The cell growth curve showed that the growth rate of cells in each exposure group was significantly slower than that in the control group. ;After 5 passages of cells exposed to the virus, ¦Â-galactosidase staining was obviously positive, while the staining of the control group was negative; cell cycle kinetics showed that with the increase of the concentration and the prolongation of the time, the G1 and G2 phases The number of cells increased and the number of cells in S phase decreased; after CSE, the expression of type ¢ñ procollagen mRNA in fibroblasts decreased. Conclusion CSE has obvious damaging effects on fibroblasts in vitro and has a certain inhibitory effect on cell proliferation and growth; the accumulation of long-term effects of low-concentration CSE makes fibroblasts that have been passaged multiple times have the characteristics of senescent cells; CSE can inhibit Cell activity causes cell DNA replication and mitosis disorders, with varying degrees of blockage in each phase. CSE has a certain inhibitory effect on collagen synthesis in fibroblasts
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Original research was done by Qian Ge; Jiang Faxing; Zhou Wuqing; Wang Qianqiu; Li Xiaowu; Zhong Mingying; Zheng Bo
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