中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2783-2785
,共3页
张华伟%李皓桓%彭飞%孙维国
張華偉%李皓桓%彭飛%孫維國
장화위%리호환%팽비%손유국
骨关节炎%软骨细胞%线粒体%胰岛素样生长因子结合蛋白-3%吡咯喹啉醌
骨關節炎%軟骨細胞%線粒體%胰島素樣生長因子結閤蛋白-3%吡咯喹啉醌
골관절염%연골세포%선립체%이도소양생장인자결합단백-3%필각규람곤
Osteoarthritis%Chondrocytes%Mitochondrion%Insulin-like growth factor binding protein-3%Pyrroloquinoline quinone
目的 观察胰岛素样生长因子结合蛋白-3(IGFBP-3)介导线粒体途径致软骨细胞凋亡机制及吡咯喹啉醌(PQQ)的保护作用.方法 不同浓度IGFBP-3或PQQ对SD乳鼠膝软骨细胞干预处理.A组为对照组;B、C、D组分别加0.5、1.0、2.0 mg/L IGFBP-3;E组:2.0 mg/L IGFBP-3+100 nmol/L PQQ.透射电镜观察线粒体超微结构,测量线粒体体密度(Vv)、表面积密度(Sv)及数密度(Nv).荧光显微镜观察线粒体膜电位.流式细胞仪检测各组细胞凋亡率.结果 透射电镜观察线粒体见A组结构正常,B、C、D组出现肿胀受损表现,E组出现局灶性空泡,但较D组明显减少.Vv、Sv和Nv发生相应改变,差异有统计学意义(P<0.05).荧光显微镜下A组呈橘红色荧光;B、C组既呈橘红色也呈绿色荧光;D组80%以上呈绿色荧光;E组可见橘红色和绿色两种荧光.A~E组橘红色荧光百分率分别为(89.7±3.4)%、(61.1±2.7)%、(46.4±3.1)%、(15.3±4.2)%和(57.8±3.4)%.细胞凋亡率A组为(2.67±0.29)%,B、C、D组分别为(7.91±0.41)%、(12.33±1.71)%和(19.56±2.62)%,与A组比较,差异有统计学意义(P<0.05);E组为(9.14±1.83)%,与D组比较差异有统计学意义(P<0.05).结论 IGFBP-3可通过线粒体途径,造成线粒体形态和功能改变,最终导致软骨细胞凋亡;而PQQ对软骨细胞线粒体具有保护作用.
目的 觀察胰島素樣生長因子結閤蛋白-3(IGFBP-3)介導線粒體途徑緻軟骨細胞凋亡機製及吡咯喹啉醌(PQQ)的保護作用.方法 不同濃度IGFBP-3或PQQ對SD乳鼠膝軟骨細胞榦預處理.A組為對照組;B、C、D組分彆加0.5、1.0、2.0 mg/L IGFBP-3;E組:2.0 mg/L IGFBP-3+100 nmol/L PQQ.透射電鏡觀察線粒體超微結構,測量線粒體體密度(Vv)、錶麵積密度(Sv)及數密度(Nv).熒光顯微鏡觀察線粒體膜電位.流式細胞儀檢測各組細胞凋亡率.結果 透射電鏡觀察線粒體見A組結構正常,B、C、D組齣現腫脹受損錶現,E組齣現跼竈性空泡,但較D組明顯減少.Vv、Sv和Nv髮生相應改變,差異有統計學意義(P<0.05).熒光顯微鏡下A組呈橘紅色熒光;B、C組既呈橘紅色也呈綠色熒光;D組80%以上呈綠色熒光;E組可見橘紅色和綠色兩種熒光.A~E組橘紅色熒光百分率分彆為(89.7±3.4)%、(61.1±2.7)%、(46.4±3.1)%、(15.3±4.2)%和(57.8±3.4)%.細胞凋亡率A組為(2.67±0.29)%,B、C、D組分彆為(7.91±0.41)%、(12.33±1.71)%和(19.56±2.62)%,與A組比較,差異有統計學意義(P<0.05);E組為(9.14±1.83)%,與D組比較差異有統計學意義(P<0.05).結論 IGFBP-3可通過線粒體途徑,造成線粒體形態和功能改變,最終導緻軟骨細胞凋亡;而PQQ對軟骨細胞線粒體具有保護作用.
목적 관찰이도소양생장인자결합단백-3(IGFBP-3)개도선립체도경치연골세포조망궤제급필각규람곤(PQQ)적보호작용.방법 불동농도IGFBP-3혹PQQ대SD유서슬연골세포간예처리.A조위대조조;B、C、D조분별가0.5、1.0、2.0 mg/L IGFBP-3;E조:2.0 mg/L IGFBP-3+100 nmol/L PQQ.투사전경관찰선립체초미결구,측량선립체체밀도(Vv)、표면적밀도(Sv)급수밀도(Nv).형광현미경관찰선립체막전위.류식세포의검측각조세포조망솔.결과 투사전경관찰선립체견A조결구정상,B、C、D조출현종창수손표현,E조출현국조성공포,단교D조명현감소.Vv、Sv화Nv발생상응개변,차이유통계학의의(P<0.05).형광현미경하A조정귤홍색형광;B、C조기정귤홍색야정록색형광;D조80%이상정록색형광;E조가견귤홍색화록색량충형광.A~E조귤홍색형광백분솔분별위(89.7±3.4)%、(61.1±2.7)%、(46.4±3.1)%、(15.3±4.2)%화(57.8±3.4)%.세포조망솔A조위(2.67±0.29)%,B、C、D조분별위(7.91±0.41)%、(12.33±1.71)%화(19.56±2.62)%,여A조비교,차이유통계학의의(P<0.05);E조위(9.14±1.83)%,여D조비교차이유통계학의의(P<0.05).결론 IGFBP-3가통과선립체도경,조성선립체형태화공능개변,최종도치연골세포조망;이PQQ대연골세포선립체구유보호작용.
Objective To evaluate the mechanism of insulin-like growth factor binding protein-3 (IGFBP-3)-mediated mitochondrial pathway in chondrocytes apoptosis and protective effect of pyrroloquinoline quinone (PQQ).Methods Chondrocytes were derived from knee of SD rats.Different concentrations of IGFBP-3 with or withnot PQQ were added into culture medium.Group A served as control group.Groups B, C, and D were given 0.5, 1.0, 2.0 mg/L IGFBP-3, respectively.Group E was given 2.0 mg/L IGFBP-3 + 100 nmol/L PQQ.The morphology of mitochondria was observed under transmission electron microscopy, then volume density (Vv), surface density (Sv) and numerical density (Nv) were measured.Mitochondrial membrane potential was assessed by fluorescent microscope.The apoptosis rate of chondrocytes was detected by flow cytometry.Results Under the electron microscope, the mitochondria morphology was normal in group A.In groups B, C and D, the swelled and vacuolated mitochondria were observed.In group E, mitochondria were irregular, and mildly variant mitochondria shown as compared with group D.The three parameters (Vv, Sv and Nv) were correspondingly changed in the different groups, and there were significant differences among the groups.Under the fluorescent microscope, orange fluorescence presented in the group A.Both orange and green fluorescence were seen in groups B and C.More than 80% green fluorescence presented in group D.Both orange and green fluorescence presented in group E.The ratio of mitochondrial orange fluorescence in groups A, B, C, D and E was (89.7 ± 3.4)%, (61.1 ±2.7)%, (46.4±3.1)%, (15.3 ±4.2)% and (57.8 ±3.4)%, respectively.Apoptosis rate in group A was (2.67 ± 0.29) %, and as compared with group A, apoptosis rate in groups B, C and D was (7.91 ± 0.41) %, (12.33 ± 1.71) % and (19.56 ± 2.62) % respectively, but that was (9.14 ± 1.83) % in group E.All differences were statistically significant.Conclusion IGFBP-3 can cause morphological and functional changes of mitochondria through mitochondrial pathway, eventually leading to cartilage cell apoptosis.The protective effects of PQQ on mitochondria in cartilage cells was verified.