中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
4期
787-789
,共3页
吡咯喹啉醌%骨关节炎%软骨细胞%线粒体
吡咯喹啉醌%骨關節炎%軟骨細胞%線粒體
필각규람곤%골관절염%연골세포%선립체
Pyrroloquinoline quinone%Osteoarthritis%Chondrocyte%Mitochondrion
目的 观察吡咯喹啉醌(PQQ)对软骨细胞线粒体超微结构及膜电位的影响.方法 体外原代培养大鼠膝关节软骨细胞,用白细胞介素(IL)-1β 10 μg/L刺激软骨细胞,同时加入不同浓度的PQQ(0、1、10、100、1000、10 000 nmol/L)共同处理24 h后观察线粒体形态学及超微结构的改变；JC-1荧光探针标记线粒体,用荧光显微镜检测线粒体膜电位的变化.结果 经IL-1β作用后,软骨细胞线粒体截面面积、截面周长和直径[(90.28±27.28) μm2、(70.47±22.88) μm、(16.83±1.13) μm]均较正常对照[(50.26±24.35) μm2、(35.28±16.67) μm、(12.16±1.51) μm]增加,线粒体出现肿胀受损表现.100 nrmol/L PQQ可拮抗IL-1β的损害作用[(74.79 ±21.54) μm2、(55.43±21.56) μm、(15.03 ±0.69) μm]；10 000 nmol/L PQQ拮抗作用减弱[(89.16±28.56) μm2、(67.39±21.77) μm、(16.51±1.51) μm]；CCP阳性对照组及A～F组橘红色线粒体荧光百分率(％)分别为(1.1±3.3、88.4±4.1、20.6 ±2.8、26.4±3.8、34.7±3.6、57.8±4.4、31.4±3.8、16.5 ±3.2),PQQ 浓度在1、10、100、1000 nmol/L时橘红色线粒体荧光百分率较IL-1β组均明显升高(P＜0.05)；PQQ 浓度为100 nmol/L时橘红色线粒体荧光百分率最高；PQQ浓度为10 000 nmol/L时对线粒体膜电位表现抑制作用.结论 PQQ对软骨细胞线粒体的结构及膜电位具有保护作用.
目的 觀察吡咯喹啉醌(PQQ)對軟骨細胞線粒體超微結構及膜電位的影響.方法 體外原代培養大鼠膝關節軟骨細胞,用白細胞介素(IL)-1β 10 μg/L刺激軟骨細胞,同時加入不同濃度的PQQ(0、1、10、100、1000、10 000 nmol/L)共同處理24 h後觀察線粒體形態學及超微結構的改變；JC-1熒光探針標記線粒體,用熒光顯微鏡檢測線粒體膜電位的變化.結果 經IL-1β作用後,軟骨細胞線粒體截麵麵積、截麵週長和直徑[(90.28±27.28) μm2、(70.47±22.88) μm、(16.83±1.13) μm]均較正常對照[(50.26±24.35) μm2、(35.28±16.67) μm、(12.16±1.51) μm]增加,線粒體齣現腫脹受損錶現.100 nrmol/L PQQ可拮抗IL-1β的損害作用[(74.79 ±21.54) μm2、(55.43±21.56) μm、(15.03 ±0.69) μm]；10 000 nmol/L PQQ拮抗作用減弱[(89.16±28.56) μm2、(67.39±21.77) μm、(16.51±1.51) μm]；CCP暘性對照組及A～F組橘紅色線粒體熒光百分率(％)分彆為(1.1±3.3、88.4±4.1、20.6 ±2.8、26.4±3.8、34.7±3.6、57.8±4.4、31.4±3.8、16.5 ±3.2),PQQ 濃度在1、10、100、1000 nmol/L時橘紅色線粒體熒光百分率較IL-1β組均明顯升高(P＜0.05)；PQQ 濃度為100 nmol/L時橘紅色線粒體熒光百分率最高；PQQ濃度為10 000 nmol/L時對線粒體膜電位錶現抑製作用.結論 PQQ對軟骨細胞線粒體的結構及膜電位具有保護作用.
목적 관찰필각규람곤(PQQ)대연골세포선립체초미결구급막전위적영향.방법 체외원대배양대서슬관절연골세포,용백세포개소(IL)-1β 10 μg/L자격연골세포,동시가입불동농도적PQQ(0、1、10、100、1000、10 000 nmol/L)공동처리24 h후관찰선립체형태학급초미결구적개변；JC-1형광탐침표기선립체,용형광현미경검측선립체막전위적변화.결과 경IL-1β작용후,연골세포선립체절면면적、절면주장화직경[(90.28±27.28) μm2、(70.47±22.88) μm、(16.83±1.13) μm]균교정상대조[(50.26±24.35) μm2、(35.28±16.67) μm、(12.16±1.51) μm]증가,선립체출현종창수손표현.100 nrmol/L PQQ가길항IL-1β적손해작용[(74.79 ±21.54) μm2、(55.43±21.56) μm、(15.03 ±0.69) μm]；10 000 nmol/L PQQ길항작용감약[(89.16±28.56) μm2、(67.39±21.77) μm、(16.51±1.51) μm]；CCP양성대조조급A～F조귤홍색선립체형광백분솔(％)분별위(1.1±3.3、88.4±4.1、20.6 ±2.8、26.4±3.8、34.7±3.6、57.8±4.4、31.4±3.8、16.5 ±3.2),PQQ 농도재1、10、100、1000 nmol/L시귤홍색선립체형광백분솔교IL-1β조균명현승고(P＜0.05)；PQQ 농도위100 nmol/L시귤홍색선립체형광백분솔최고；PQQ농도위10 000 nmol/L시대선립체막전위표현억제작용.결론 PQQ대연골세포선립체적결구급막전위구유보호작용.
Objective To investigate the effect of pyrroloquinoline quinine (PQQ) on mitochondria ultrastructure and mitochondrial membrane potential (△Ψm) of chondrocytes.Methods The chondrocytes derivel from the knee of rats were subjected to primary culture in vitro.After identification by toluidine blue staining,10 μg/L interleukin (IL)-1 β in the presence or absence of PQQ with varied concentrations (0,1,10,100,1000 and 10 000 nmol/L) was added into culture medium and co-cultured for 24 h.The morphology of mitochondria was observed under the transmission electron microscopy (TEM) and was quantitatively analyzed by a MiVnt image analysis system,and /Ψm was assessed by JC-1 staining under fluorescent microscope.Results After IL-1β treatment,the mitochondrial areas,perimeters and diameters of chondrocytes [(90.28 ±27.28) μm2,(70.47 ±22.88) μm,(16.83 ± 1.13) μm] were increased as compared with control group [(50.26 ±24.35) μm2,(35.28 ± 16.67) μm,(12.16 ± 1.51) μm],which presented a damage appearance.100 nmol/L PQQ could antagonist the effect [(74.79±21.54) μm2,(55.43 ±21.56) μm,(15.03 ±0.69) μm],but the protective effect decreased at 10 000 nmol/L PQQ [(89.16 ±28.56) μm2,(67.39 ±21.77) μm,(16.51 ± 1.51) μm] ;the mean of fluorescence intensity of JC-1 with the chondrocyte mitochondria and the percentage of orangered colored mitochondria in different concentrations of PQQ (0,1,10,100,1000 and 10 000 nmol/L)were increased significantly (1.1 ± 3.3,88.4 ± 4.1,20.6 ± 2.8,26.4 ± 3.8,34.7 ± 3.6,57.8 ± 4.4,31.4 ±3.8,16.5 ±3.2).The maximal effect occurred on 100 nmol/L PQQ,and 10 000 nmol/L PQQ exhibited the depressed effect on morphology of mitochondria on cultured chondrocytes (P ＜ 0.05).Conclusion The mitochondrial structure and △ Ψm of the chondrocytes were protected by PQQ.
