内毒素脂多糖促进人牙髓成纤维细胞凋亡及其对caspase-3活性的影响  

Experimental research of apoptosis in dental pulp cells induced by LPS and effects on activity of caspase-3

作　　者：戴秀峰[1] 吴平[2] 何惠娟[2] 杨展[2] 王旭光[2] 陈坤[1] 

机构地区：[1]广东医学院附属医院口腔科,湛江524001 [2]广东医学院附属医院中心实验室,湛江524001

出　　处：《口腔医学》2008年第7期358-360,363,共4页Stomatology

摘　　要：目的观察内毒素脂多糖(lipopolysaccharide,LPS)对体外培养的人牙髓成纤维细胞的致凋亡作用及其对凋亡相关的半胱氨酸蛋白酶3(caspase-3)活性的影响。方法不同浓度(0μg、100μg、200μg、400μg/mL)的LPS作用细胞,采用噻唑蓝比色法检测LPS对细胞增殖的影响;Hoechst33258荧光染色法观察细胞凋亡;流式细胞术检测细胞周期及凋亡;化学比色法检测细胞凋亡过程中caspase-3活性的变化。结果不同浓度LPS组细胞的生长与对照组相比均受到显著的抑制(P<0.01),细胞增殖抑制率呈剂量依赖关系;Hoechst33258荧光染色显示,给药组部分细胞呈典型的凋亡表现;流式细胞术检测结果显示,不同浓度的LPS作用细胞48h,细胞的凋亡率分别为9.2%,23.8%,31.4%;caspase-3活性检测显示,其活性呈剂量依赖性增高,LPS浓度达到400μg/mL时,其活性为对照组的3.4倍。结论LPS可显著地抑制人牙髓成纤维细胞增殖、诱导细胞凋亡、升高caspase-3活性,并呈明显的量效关系。Objective To observe the induced apoptosis effect of Lipopolysaccharide（LPS） on human dental pulp body fibroblast（HDF） cultured in vitro and the activity of caspase-3. Method Different concentrations of LPS（0μg/mL, 100μg/mL, 200μg/mL, 400μg/mL） were used to treat the cells. Thiazole blue chromatomentry was employed to detect the effects of LPS on cellpmliferation. When Hoechst.33258 fluorescent staining cell apoptosis was observed, cytometry was flown to detect cell cycle and apoptosis. Chemistry chromatometry was adopted to detect the change of caspase-3＇s activity during the process of cell apoptosis. Result Compared with the control group, the cell growth in the experimental group was depressed significantly（ P 〈 0.01 ）, and the cellproliferation depressed rate showed the dependent relation to the dosage. Hoechst33258 fluorescent staining displayed that cells in the experimental group partly showed typical apoptosis. Flow cytometrie analyses showed that the apoptosis rates were 9.2 %, 23.8 % and 31.4 %, respectively, after the cells were treated with different concentrations of LPS for 48h. The dection of caspase-3 activity showed that the activity increased dependently on dosage. The activity was 3.4 times of the control group when the concentration of LPS was 400μg/mL. Conclusion LPS can depress the proliferation of PDF, induce cell apoptosis, increase the activity of caspase - 3, and shows apparent relation with dosaqes.

关 键 词：内毒素脂多糖 牙髓细胞 凋亡 半胱氨酸蛋白酶-3 

分 类 号：R349.53[医药卫生—基础医学]