Inhibitory effect of recombinant human endostatin on the proliferation of hypertrophic scar fibroblasts in a rabbit ear model.
Hypertrophic scar (HS) is a pathological scar that particularly occurs after traumatic injuries, surgical procedures and burning. Abnormal activation of hypertrophic scar fibroblasts (HSFs) intensifies fibrosis during wound healing. Our previous studies demonstrated that recombinant human endostatin (rhEndostatin) prevented synovial thickening in adjuvant arthritis (AA) rats via inhibition of proliferation and enhancement of apoptosis in synovial fibroblasts. However, the effect of this protein on HSF proliferation is not known. This study investigated the inhibitory effect of rhEndostatin on the proliferation of cultured HSFs in a rabbit ear model. MTT assay and flow cytometric detection were performed to investigate HSF proliferation and cell cycle progression, respectively. The expression levels of p53, p21, cyclinD1, cyclin-dependent kinase 4 (CDK4) and proliferating cell nuclear antigen (PCNA) in HSFs were detected using real-time PCR and Western blotting. Our data revealed that HSFs treated with rhEndostatin were significantly inhibited in a concentration-dependent manner with an IC50value of 100mg/L. Also, rhEndostatin (100mg/L) primarily induced G0/G1and partially G2/M cell cycle arrest of HSFs. There were significant decreases in the expression levels of p53, p27, CDK4, cyclinD1and PCNA in HSFs treated with rhEndostatin. In conclusion, rhEndostatin inhibited HSF proliferation via G0/G1and/or G2/M phase arrest of the cell cycle, which was partially due to the down-regulation of cyclinD1, CDK4and PCNA. These findings suggest that rhEndostatin may reduce scar hypertrophy in vivo via inhibition of HSF proliferation and may be a novel agent for HS treatment.
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: gyfanhui@126.com.
Yong-Fang
YF
Gong
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: zhxm_1919@126.com.
Xiao-Ming
XM
Zhang
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: liufei1016@163.com.
Fei
F
Liu
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: wangzz190@gmail.com.
Zhen-Zhen
ZZ
Wang
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: knowou@163.com.
Xue-Fei
XF
Deng
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: jiaoyi7721@aliyun.com.
Yi
Y
Jiao
Department of Plastic Surgery, The First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei 230022, China. Electronic address: ahzyangel@sina.com.
Xiao-Jing
XJ
Li
Department of Anatomy, Anhui Medical University, 81 Meishan Road, Hefei 230032, China. Electronic address: hxy8676@126.com.
Xue-Ying
XY
Huang
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D017439:Cicatrix, Hypertrophic / Q000188:drug therapy / Q000473:pathology*; D004195:Disease Models, Animal; D004423:Ear / Q000473:pathology*; D043169:Endostatins / Q000494:pharmacology* / Q000627:therapeutic use; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology* / Q000627:therapeutic use
D043169:Endostatins; D011994:Recombinant Proteins
Fibroblast; Hypertrophic scar; Proliferation; Recombinant human endostatin