摘要
目的体外模拟成骨细胞在体内的生存环境,探讨淫羊藿苷(ICA)对高重力下成骨细胞MC3T3-E1增殖与凋亡的影响。方法将成骨细胞随机分为三组:高重力组、高重力+ICA组(以下简称"联合组")和对照组,其中,高重力组予以20G高重力,联合组将20G高重力和10μg/L ICA同时应用于细胞,对照组正常培养,不进行任何附加操作。采用MTT法检测细胞增殖,流式细胞术(FCM)检测细胞凋亡,实时定量PCR(qRT-PCR)检测细胞凋亡相关基因,透射电镜(TEM)检测细胞亚细胞结构变化。结果 MTT法检测显示,高重力组吸光度值显著低于对照组(P<0.01),联合组吸光度值显著高于对照组和高重力组(P<0.01)。FCM检测显示,高重力组凋亡率显著高于对照组(P<0.01),联合组凋亡率显著低于对照组和高重力组(P<0.01)。qRT-PCR检测结果显示,与对照组比较,高重力组Bax、Bcl2、Bax/Bcl2、P53的mRNA表达均显著上调(P<0.01或P<0.05),联合组除Bax/Bcl2 mRNA表达外均显著下调(P<0.05或P<0.01),且联合组Bax、Bcl2、Bax/Bcl2、P53的mRNA表达均显著低于高重力组(均P<0.01)。TEM下可见高重力组呈中晚期凋亡和变性坏死现象,而其他两组无明显变化。结论成骨细胞可响应高重力刺激信号并发生一系列变化;高重力(20G)不仅可抑制成骨细胞增殖,促进细胞凋亡,还可导致成骨细胞发生变性坏死,但适当浓度(10μg/L)的ICA可逆转高重力加载所导致的这种不利影响,对成骨细胞起相应的保护作用。
Objective To investigate the effects of icariin （ICA） on the proliferation and apoptosis of osteoblasts MC3T3-E1 under hypergravity by simulating the survival environment of osteoblasts in vivo. Methods The osteoblasts were randomly divided into three groups： hypergravity group, bypergravity＋ICA group （＂combined group＂ for short） and control group. The hypergravity group was treated with 20G hypergravity; in combined group, 20G hypergravity and 10 μg/L ICA were applied to the cells; the control group was cultured without any additional operation. Cell proliferation was detected by MTT assay, the apoptosis was detected by flow cytometry （FCM）, the expression of apoptosis related genes was detected by quantitative real-time PCR （qRT-PCR）, and the subcellular structure was detected by transmission electron microscope （TEM）. Results The result of MTT examination showed that the absorbance value of hypergravity group was significantly lower than that of control group （P 〈 0.01）, while the absorbance value of combined group was significantly higher than those of control group and hypergravity group （P 〈 0.01）. FCM method showed that the apoptosis rate of hypergravity group was significantly higher than that of control group （P 〈 0.01）, while the apoptosis rate of combined group was significantly lower than those of control group and hypergravity group （P 〈 0.01）. qRT-PCR showed that, compared with control group, the mRNA expression of Bax, Bcl2, Bax/Bcl2, P53 in hypergravity group was significantly increased （P 〈 0.01 or P 〈 0.05）, but in combined group, the mRNA expression above was significantly decreased except Bax/Bcl2 mRNA （P 〈 0.05 or P 〈 0.01）, and the mRNA expression of Bax, Bcl2, Bax/Bcl2, P53 in combined group was significantly lower than those in hypergravity group （P 〈 0.01）. TEM indicated that the hypergravity group showed advanced apoptosis and degeneration necrosis, but there were no significant changes in the other two groups. Conclusion Osteoblasts can respond to hypergravity stimulation and appear a series of changes. Hypergravity （20G） can not only inhibit osteoblasts＇ proliferation and promote apoptosis, but also lead to degeneration and necrosis of osteoblasts. But the appropriate concentration （10 μg/L） ICA can protect osteoblasts from the adverse effect caused by hypergravity load.
作者
李军
宋光明
孙明林
黃揆
张春秋
付浩
李瑞欣
张西正
LI Jun SONG Guangming SUN Minglin HUANG Kui ZHANG Chunqiu FU Hao LI Ruixin ZHANG Xiz he ng(Department of Spine, Affiliated Hospital＂ of Logistics University of People＇s Armed Police Force, Tianjin 300162, China Staff Room of Pharmacology, Logistics University of People＇s Armed Police Force, Tianjin 300309, China Institute of Medical Equipment, the Academy of Military Medical Sciences, Tianjin 300161, China Tianjin Key Laboratory for Advanced Mechatronic System Design and Intelligent Control, Tianjin 300384, China)
出处
《中国医药导报》
CAS
2017年第6期19-23,共5页
China Medical Herald
基金
国家自然科学基金资助重点项目(11432016)
国家自然科学基金资助面上项目(31370942)
关键词
成骨细胞
淫羊藿苷
高重力
增殖
凋亡
Osteoblast
Icariin
Hypergravity
Proliferation
Apoptosis
