中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2011年
2期
130-133
,共4页
张铁山%王文波%宋玉文%刘贾昆
張鐵山%王文波%宋玉文%劉賈昆
장철산%왕문파%송옥문%류가곤
氟%软骨细胞%基因,bcl-2%bcl-2相关X蛋白质
氟%軟骨細胞%基因,bcl-2%bcl-2相關X蛋白質
불%연골세포%기인,bcl-2%bcl-2상관X단백질
Fluorine%Chondrocytes%Genes,bcl-2%bcl-2-associated X protein
目的 观察氟对体外培养软骨细胞中凋亡相关因子bcl-2和Bax蛋白表达的影响,探讨氟在致软骨细胞凋亡中的作用机制.方法 采用软骨细胞体外培养方法,原代培养乳鼠关节软骨细胞,传第3代后按染氟剂量不同分为0(对照)、5、20、40mg/L组,培养10 d后,透射电镜下观察软骨细胞的超微结构改变,采用Western印迹法检测软骨细胞bcl-2和Bax蛋白表达.结果 透射电镜下,对照组和5 mg/L组软骨细胞呈球形,粗面内质网发达,线粒体膜性结构完整;20、40 mg/L组软骨细胞内可见脂滴增多,胞质内出现大量空泡类物质,细胞内膜结构不清,部分细胞出现核固缩.20、40 mg/L组软骨细胞bcl-2蛋白表达(0.626±0.042、0.531±0.039)较对照组(0.876±0.035)明显降低(P均＜0.01);而Bax蛋白表达(0.966±0.047、1.289±0.156)较对照组(0.642±0.050)明显增加(P均＜0.01).5 mg/L组软骨细胞bcl-2、Bax蛋白表达(0.885±0.065、0.657±0.045)与对照组比较,差异无统计学意义(P均＞0.05).此外,40 mg/L组与20 mg/L组比较,bcl-2、Bax蛋白表达差异有统计学意义(P均＜0.01).结论 染氟20、40 mg/L可对软骨细胞超微结构造成损伤,其通过减少抑凋亡因子bcl-2的表达和增加促凋亡因子Bax的表达,从而产生促进软骨细胞凋亡的作用.
目的 觀察氟對體外培養軟骨細胞中凋亡相關因子bcl-2和Bax蛋白錶達的影響,探討氟在緻軟骨細胞凋亡中的作用機製.方法 採用軟骨細胞體外培養方法,原代培養乳鼠關節軟骨細胞,傳第3代後按染氟劑量不同分為0(對照)、5、20、40mg/L組,培養10 d後,透射電鏡下觀察軟骨細胞的超微結構改變,採用Western印跡法檢測軟骨細胞bcl-2和Bax蛋白錶達.結果 透射電鏡下,對照組和5 mg/L組軟骨細胞呈毬形,粗麵內質網髮達,線粒體膜性結構完整;20、40 mg/L組軟骨細胞內可見脂滴增多,胞質內齣現大量空泡類物質,細胞內膜結構不清,部分細胞齣現覈固縮.20、40 mg/L組軟骨細胞bcl-2蛋白錶達(0.626±0.042、0.531±0.039)較對照組(0.876±0.035)明顯降低(P均＜0.01);而Bax蛋白錶達(0.966±0.047、1.289±0.156)較對照組(0.642±0.050)明顯增加(P均＜0.01).5 mg/L組軟骨細胞bcl-2、Bax蛋白錶達(0.885±0.065、0.657±0.045)與對照組比較,差異無統計學意義(P均＞0.05).此外,40 mg/L組與20 mg/L組比較,bcl-2、Bax蛋白錶達差異有統計學意義(P均＜0.01).結論 染氟20、40 mg/L可對軟骨細胞超微結構造成損傷,其通過減少抑凋亡因子bcl-2的錶達和增加促凋亡因子Bax的錶達,從而產生促進軟骨細胞凋亡的作用.
목적 관찰불대체외배양연골세포중조망상관인자bcl-2화Bax단백표체적영향,탐토불재치연골세포조망중적작용궤제.방법 채용연골세포체외배양방법,원대배양유서관절연골세포,전제3대후안염불제량불동분위0(대조)、5、20、40mg/L조,배양10 d후,투사전경하관찰연골세포적초미결구개변,채용Western인적법검측연골세포bcl-2화Bax단백표체.결과 투사전경하,대조조화5 mg/L조연골세포정구형,조면내질망발체,선립체막성결구완정;20、40 mg/L조연골세포내가견지적증다,포질내출현대량공포류물질,세포내막결구불청,부분세포출현핵고축.20、40 mg/L조연골세포bcl-2단백표체(0.626±0.042、0.531±0.039)교대조조(0.876±0.035)명현강저(P균＜0.01);이Bax단백표체(0.966±0.047、1.289±0.156)교대조조(0.642±0.050)명현증가(P균＜0.01).5 mg/L조연골세포bcl-2、Bax단백표체(0.885±0.065、0.657±0.045)여대조조비교,차이무통계학의의(P균＞0.05).차외,40 mg/L조여20 mg/L조비교,bcl-2、Bax단백표체차이유통계학의의(P균＜0.01).결론 염불20、40 mg/L가대연골세포초미결구조성손상,기통과감소억조망인자bcl-2적표체화증가촉조망인자Bax적표체,종이산생촉진연골세포조망적작용.
Objective To study the effect of fluoride on the expression of bcl-2 and Bax in chondrocyte in vitro, and investigate the mechanism of action of chondrocyte apoptosis induced by fluoride. Methods Articular chondrocytes of neonate rat were cultured in vitro and treated with 0(control),5,20,40 mg/L of fluoride,respectively, for 10 days. Then observed the u]trastructure of chondrocytes under eletronicmicroscope, and tested the expression of bcl-2 and Bax in chondrocyte in different groups by Western blotting. Results Abundant rough endoplasmic reticulums (RERs) and complete structure of mitochondria membranes were presented in globular chondrocytes in the control group and 5 mg/L group; but more lipid droplets and vacuoles were seen in the cytoplasm, and the structure of intracellular membranes became incomplete, and some shrieked chromatin and pyknosis were seen in the chondrocytes of the 20,40 mg/L groups. The expression of bcl-2 markedly decreased in 20 mg/L group(0.626 ± 0.042) and 40 mg/L group(0.531± 0.039) compared to the control group(0.876 ± 0.035,all P ＜ 0.01 ). And the expression of Bax significantly increased in 20 mg/L group(0.966 ± 0.047) and 40 mg/Lgroup ( 1 .289 ± 0.156) compared to the control group(0.642 ± 0.050, all P ＜ 0.01). But there was no statistical significant difference of the expression of bcl-2 or Bax between 5 mg/L group(0.885 ± 0.065,0.657 ± 0.045) and control group (all P ＞ 0.05 ). However there were statistical differences of expressions of bcl-2 and Bax between 20 and 40 mg/L groups(all P ＜ 0.01 ). Conclusions Twenty and 40 mg/L fluoride can cause damage to the ultrastructure of chondrocyte, and fluoride possibly promotes chondrocyte apoptosis by reducing the expression of antiapoptotic factor bcl-2 and increasing the expression of Bax.
