Basic & Clinical Medicine ›› 2020, Vol. 40 ›› Issue (3): 328-333.

• Original Articles • Previous Articles     Next Articles

Down-regulation of Notch1 inhibits proliferation and induces apoptosis of fibroblasts from pathological scars

SU Jiang-wei, YU Hui*, DU Kun, WANG Jie, LIU Lin, CAI Jian   

  1. Department of Dermatology, Ezhou Central Hospital, Ezhou 436000, China
  • Received:2019-04-22 Revised:2019-11-18 Online:2020-03-05 Published:2020-03-02
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Abstract: Objective To investigate the effect of down-regulation of Notch1 expression mediated by lentivirus on the activation of caspase-9 and mitochondrial membrane potential of fibroblast from pathological scars(PSF). Methods PSFs were isolated and cultured and then infected by Notch1 siRNA lentivirus and negative control lentivirus respectively.The level of Notch1 mRNA and protein was detected by real-time PCR and Western blot,respectively.MTT assay was used to detect cell proliferation; PI monochrome staining was used to detect cell cycle changes. Annexin V-FITC/PI assay was used to detect apoptotic changes; Western blot was used to detect the level of cyclin D1, CDK4, c-caspase-3, c-caspase-9 and cytochrome C protein levels in cytoplasm and mitochondria; JC-1 method was used to detect the mitochondrial membrane potential. Results The expression of Notch1 in PSFs transfected with Notch1 siRNA lentivirus was significantly lower than that in the negative control lentiviruses and the non-transfected PSFs(P<0.05).The proliferation of PSFs decreased after Notch1 was down-regulated (P<0.05);the percen-tage of G0/G1 phase increased(P<0.05); the apoptotic rate increased(P<0.05); the expression of c-caspase-3 and c-caspase-9 protein increased(P<0.05); the mitochondrial membrane potential of the cells decreased significantly(P<0.05); the level of cyclin D1 and CDK4 protein in cells decreased(P<0.05); cytochrome C protein increased in cytoplasm and decreased in mitochondria(P<0.05). Conclusions Down-regulation of Notch1 inhibits proliferation and induces apoptosis of PSF.

Key words: Notch1, fibroblast from pathological scars, mitochondrial membrane potential, caspase

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