[热损伤角质形成细胞上清液对真皮成纤维细胞Ⅰ型胶原、Ⅲ型胶原和基质金属蛋白酶1表达的影响]
[Effects of heat injured keratinocytes supernatant on expressions of collagen type I, collagen type III, and matrix metalloproteinase 1 of dermal fibroblasts].
作者信息
Yang Xuekang, Bai Xiaozhi, Zhang Yue, Li Xiaoqiang, Shi Jihong, Tang Chaowu, Hu Dahai
机构信息
Burn Center of Chinese PLA, Xijing Hospital, the Fourth Military Medical University, Xi'an Shaanxi, 710032, P.R.China.
出版信息
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2012 Dec;26(12):1497-500.
OBJECTIVE
To investigate the effects of heat injured keratinocytes (KC) supernatant on the expressions of collagen type I, collagen type III, and matrix metalloproteinase 1 (MMP-1) of dermal fibroblasts (Fb).
METHODS
KC and Fb were isolated and cultured. Then the models of heat injured KC and Fb were reproduced in vitro, respectively. The heat injured and normal culture supernatant were collected respectively at 12 hours, and formulated as a 50% concentration of cell-conditioned medium. According to the culture medium, Fb at passage 3-5 was divided into 3 groups. Normal Fb was cultured with the conditioned medium containing 50% heat injured KC culture supernatant (group A), the conditioned medium containing 50% normal KC culture supernatant (group B), and DMEM (group C), respectively. The cells in 3 groups were collected at 24 hours. In addition, the cells in group A were collected at 0, 1, 2, 6, 12, 24, and 48 hours, respectively. Normal Fb was cultured with the conditioned medium containing 50% heat injured Fb culture supernatant. Then, the cells were collected at 0, 1, 2, 6, 12, 24, and 48 hours, respectively. The mRNA levels of the collagen type I, collagen type III, and MMP-1 of Fb were measured by real-time fluorescent quantitative PCR techniques.
RESULTS
At 24 hours after cultured with supernatant of heat injured KC, mRNA relative expression levels of collagen type I, collagen type III, and MMP-1 in group A were significantly higher than those in groups B and C (P < 0.05). The mRNA relative expression levels of collagen type I, collagen type III, and MMP-1 in group A gradually increased with time going, showing significant differences between 0 hour and 2, 6, 12, 24, and 48 hours (P < 0.05); significant differences were found between different time points after 2 hours (P < 0.05). After Fb was treated with supernatant of heat injured Fb, the mRNA relative expression levels of MMP-1 gradually decreased with time going, showing significant differences between 0 hour and 1, 2, 6, 12, 24, and 24 hours (P < 0.05); after 2 hours of culture, significant differences were found among different time points (P < 0.05).
CONCLUSION
Heat injured KC supernatant may regulate the mRNA expressions of collagen type I, collagen type III, and MMP-1 of Fb.
目的
探讨热损伤角质形成细胞（KC）上清液对真皮成纤维细胞（Fb）Ⅰ型胶原、Ⅲ型胶原及基质金属蛋白酶1（MMP-1）表达的影响。
方法
分离培养KC和Fb，分别在体外建立热损伤KC和Fb模型。于12小时分别收集热损伤组和正常培养组的上清液，配制成50%浓度的细胞条件培养基。根据培养基不同，将第3-5代Fb分为3组，正常Fb分别用含50%热损伤KC培养上清液的条件培养基（A组）、含50%正常KC培养上清液的条件培养基（B组）和DMEM（C组）培养。24小时收集3组细胞，另外，A组细胞分别在0、1、2、6、12、24和48小时收集。正常Fb用含50%热损伤Fb培养上清液的条件培养基培养，然后分别在0、1、2、6、12、24和48小时收集细胞。采用实时荧光定量PCR技术检测Fb中Ⅰ型胶原、Ⅲ型胶原及MMP-1的mRNA水平。
结果
用热损伤KC上清液培养24小时后，A组中Ⅰ型胶原、Ⅲ型胶原及MMP-1的mRNA相对表达水平显著高于B组和C组（P<0.05）。A组中Ⅰ型胶原、Ⅲ型胶原及MMP-1的mRNA相对表达水平随时间逐渐升高，在0小时与2、6、12、24和48小时之间差异有统计学意义（P<0.05）；2小时后不同时间点之间差异有统计学意义（P<0.05）。Fb用热损伤Fb上清液处理后，MMP-1的mRNA相对表达水平随时间逐渐降低，在0小时与1、2、6、12、24和48小时之间差异有统计学意义（P<0.05）；培养2小时后，不同时间点之间差异有统计学意义（P<0.05）。
结论
热损伤KC上清液可能调控Fb中Ⅰ型胶原、Ⅲ型胶原及MMP-1的mRNA表达。
Zotero 插件