中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2010年
5期
480-484
,共5页
段琛%郭雄%张晓东%高宗强%张银刚%于曰祥
段琛%郭雄%張曉東%高宗彊%張銀剛%于曰祥
단침%곽웅%장효동%고종강%장은강%우왈상
大骨节病%硒%软骨细胞%细胞凋亡
大骨節病%硒%軟骨細胞%細胞凋亡
대골절병%서%연골세포%세포조망
Kaschin-Beck disease%Selenium%Chondrocytes%Apoptosis
目的 观察硒对体外培养大骨节病(KBD)患者和正常人关节软骨细胞增殖和凋亡的影响,探索补硒防治KBD的作用,并为硒对正常软骨细胞生长的影响提供依据.方法 依据<大骨节病临床诊断标准>(GB 16003-1995),选择Ⅱ度和Ⅲ度KBD患者5例和非病区正常人意外事故者5例的关节软骨进行体外分离、培养.KBD组和对照组分别给予不同剂量的硒(0、0.0125、0.0250、0.0500、0.1000、0.2500、0.5000、1.0000 mg/L)进行干预,采用四氮唑蓝(MTT)法、流式细胞仪和免疫组化法观察细胞生长和凋亡情况.结果 对照组第6天时各剂量组的细胞增殖率(0.086±0.025、0.077±0.012、0.073±0.027、0.071±0.017、0.058±0.028、0.052±0.028和0.046±0.037)比0 mg/L组(0.138±0.026)明显降低(P均＜0.05);0.1000～1.0000 mg/L剂量组的平均细胞增殖率为负值(-0.001±0.001、-0.003±0.000、-0.003±0.001和-0.004±0.001),显著低于0 mg/L组(0.025±0.003,P均＜0.05);KBD组,与0 mg/L组(0.115±0.011)比较,0.2500 mg/L剂量组促进细胞增殖(0.128±0.037,P＜0.05),1.0000 mg/L剂量组细胞生长受到抑制(0.071±0.019,P＜0.05).对照组0.0500～1.0000mg/L剂量组的细胞凋亡率[(18.88±0.02)%、(17.58±0.01)%、(17.09±0.04)%、(56.00±0.02)%、(57.85±0.03)%]比0 mg/L组[(13.51±0.01)%]增高(P均＜0.05);KBD组,与0 mg/L组[(25.84±0.02)%]比较,0.0250～0.2500 mg/L剂量组的细胞凋亡率[(13.69±0.02)%、(15.96±0.03)%、(16.68±0.03)%、(16.67±0.02)%]降低,0.5000、1.0000 mg/L剂量组的细胞凋亡率[(59.58±0.03)%、(73.48±0.04)%]明显增高(P均＜0.05).KBD组0.0500～0.2500 mg/L剂量组的Fas表达[(41.2±1.5)%、(40.3±2.0)%、(50.2±2.5)%]低于同剂量硒干预的对照组[(52.4±1.0)%、(67.2±4.0)%、(75.1±5.0)%,P均＜0.05],0.0500、0.1000 mg/L剂量组的Caspase-3表达[(40.8±1.1)%、(45.1±2.1)%]低于同剂量硒干预的对照组[(68.0±3.0)%、(70.6±3.5)%,P均＜0.05].结论 适宜的补硒剂量(0.1000～0.2500 mg/L)具有促进KBD软骨细胞生长的作用,降低细胞凋亡率,但补硒剂量＞0.5000 mg/L时具有损伤作用;促进KBD软骨细胞生长的硒剂量并非也能促进正常人活体软骨细胞的生长.
目的 觀察硒對體外培養大骨節病(KBD)患者和正常人關節軟骨細胞增殖和凋亡的影響,探索補硒防治KBD的作用,併為硒對正常軟骨細胞生長的影響提供依據.方法 依據<大骨節病臨床診斷標準>(GB 16003-1995),選擇Ⅱ度和Ⅲ度KBD患者5例和非病區正常人意外事故者5例的關節軟骨進行體外分離、培養.KBD組和對照組分彆給予不同劑量的硒(0、0.0125、0.0250、0.0500、0.1000、0.2500、0.5000、1.0000 mg/L)進行榦預,採用四氮唑藍(MTT)法、流式細胞儀和免疫組化法觀察細胞生長和凋亡情況.結果 對照組第6天時各劑量組的細胞增殖率(0.086±0.025、0.077±0.012、0.073±0.027、0.071±0.017、0.058±0.028、0.052±0.028和0.046±0.037)比0 mg/L組(0.138±0.026)明顯降低(P均＜0.05);0.1000～1.0000 mg/L劑量組的平均細胞增殖率為負值(-0.001±0.001、-0.003±0.000、-0.003±0.001和-0.004±0.001),顯著低于0 mg/L組(0.025±0.003,P均＜0.05);KBD組,與0 mg/L組(0.115±0.011)比較,0.2500 mg/L劑量組促進細胞增殖(0.128±0.037,P＜0.05),1.0000 mg/L劑量組細胞生長受到抑製(0.071±0.019,P＜0.05).對照組0.0500～1.0000mg/L劑量組的細胞凋亡率[(18.88±0.02)%、(17.58±0.01)%、(17.09±0.04)%、(56.00±0.02)%、(57.85±0.03)%]比0 mg/L組[(13.51±0.01)%]增高(P均＜0.05);KBD組,與0 mg/L組[(25.84±0.02)%]比較,0.0250～0.2500 mg/L劑量組的細胞凋亡率[(13.69±0.02)%、(15.96±0.03)%、(16.68±0.03)%、(16.67±0.02)%]降低,0.5000、1.0000 mg/L劑量組的細胞凋亡率[(59.58±0.03)%、(73.48±0.04)%]明顯增高(P均＜0.05).KBD組0.0500～0.2500 mg/L劑量組的Fas錶達[(41.2±1.5)%、(40.3±2.0)%、(50.2±2.5)%]低于同劑量硒榦預的對照組[(52.4±1.0)%、(67.2±4.0)%、(75.1±5.0)%,P均＜0.05],0.0500、0.1000 mg/L劑量組的Caspase-3錶達[(40.8±1.1)%、(45.1±2.1)%]低于同劑量硒榦預的對照組[(68.0±3.0)%、(70.6±3.5)%,P均＜0.05].結論 適宜的補硒劑量(0.1000～0.2500 mg/L)具有促進KBD軟骨細胞生長的作用,降低細胞凋亡率,但補硒劑量＞0.5000 mg/L時具有損傷作用;促進KBD軟骨細胞生長的硒劑量併非也能促進正常人活體軟骨細胞的生長.
목적 관찰서대체외배양대골절병(KBD)환자화정상인관절연골세포증식화조망적영향,탐색보서방치KBD적작용,병위서대정상연골세포생장적영향제공의거.방법 의거<대골절병림상진단표준>(GB 16003-1995),선택Ⅱ도화Ⅲ도KBD환자5례화비병구정상인의외사고자5례적관절연골진행체외분리、배양.KBD조화대조조분별급여불동제량적서(0、0.0125、0.0250、0.0500、0.1000、0.2500、0.5000、1.0000 mg/L)진행간예,채용사담서람(MTT)법、류식세포의화면역조화법관찰세포생장화조망정황.결과 대조조제6천시각제량조적세포증식솔(0.086±0.025、0.077±0.012、0.073±0.027、0.071±0.017、0.058±0.028、0.052±0.028화0.046±0.037)비0 mg/L조(0.138±0.026)명현강저(P균＜0.05);0.1000～1.0000 mg/L제량조적평균세포증식솔위부치(-0.001±0.001、-0.003±0.000、-0.003±0.001화-0.004±0.001),현저저우0 mg/L조(0.025±0.003,P균＜0.05);KBD조,여0 mg/L조(0.115±0.011)비교,0.2500 mg/L제량조촉진세포증식(0.128±0.037,P＜0.05),1.0000 mg/L제량조세포생장수도억제(0.071±0.019,P＜0.05).대조조0.0500～1.0000mg/L제량조적세포조망솔[(18.88±0.02)%、(17.58±0.01)%、(17.09±0.04)%、(56.00±0.02)%、(57.85±0.03)%]비0 mg/L조[(13.51±0.01)%]증고(P균＜0.05);KBD조,여0 mg/L조[(25.84±0.02)%]비교,0.0250～0.2500 mg/L제량조적세포조망솔[(13.69±0.02)%、(15.96±0.03)%、(16.68±0.03)%、(16.67±0.02)%]강저,0.5000、1.0000 mg/L제량조적세포조망솔[(59.58±0.03)%、(73.48±0.04)%]명현증고(P균＜0.05).KBD조0.0500～0.2500 mg/L제량조적Fas표체[(41.2±1.5)%、(40.3±2.0)%、(50.2±2.5)%]저우동제량서간예적대조조[(52.4±1.0)%、(67.2±4.0)%、(75.1±5.0)%,P균＜0.05],0.0500、0.1000 mg/L제량조적Caspase-3표체[(40.8±1.1)%、(45.1±2.1)%]저우동제량서간예적대조조[(68.0±3.0)%、(70.6±3.5)%,P균＜0.05].결론 괄의적보서제량(0.1000～0.2500 mg/L)구유촉진KBD연골세포생장적작용,강저세포조망솔,단보서제량＞0.5000 mg/L시구유손상작용;촉진KBD연골세포생장적서제량병비야능촉진정상인활체연골세포적생장.
Objective To investigate the effect of selenium on proliferation and apoptosis of chondrocytes of articular cartilage cultured in vitro in Kaschin-Beck disease(KBD) patients and normal person, to explore the role of selenium in control of KBD, and to provide evidence for selenium's effect on the growth of normal cartilage cells. Methods The articular cartilage samples of grade Ⅱ and Ⅲ KBD patients were selected according to the national "Clinical Diagnosis of KBD" (GB 16003-1995). Chondrocytes of 5 KBD and 5 non-endemic normal accidentswere separated and cultured in vitro. KBD group and control group were given different doses of selenium (0,0.0125,0.0250,0.0500,0.1000,0.2500,0.5000,1.0000 mg/L, respectively). Methyl thiazolyl tetrazolium (MTT),flow cytometric analysis, and immunocytochemical staining were used to observe the effect of selenium on cell growth and apoptosis in KBD and normal persons. Results MTT results showed that the cell proliferation rate in each dosage group of the control group at the 6th day(0.086 ± 0.025,0.077 ± 0.012,0.073 ± 0.027,0.071 ± 0.017,0.058 ± 0.028,0.052 ± 0.028 and 0.046 ± 0.037) was significantly lower than that of 0 mg/L group(0.138 ± 0.026,all P ＜ 0.05);the average cell proliferation rate was negative( - 0.001 ± 0.001, - 0.003 ± 0.000, - 0.003 ± 0.001and - 0.004 ± 0.001 ) in 0.1000 - 1.0000 mg/L dose group, which was significantly lower than that of the 0 mg/L group(0.025 ± 0.003, all P ＜ 0.05);compared with 0 mg/L group(0. 115 ± 0.011), the KBD 0.2500 mg/L dose group promoted cell proliferation(0.128 ± 0.037, P ＜ 0.05), the KBD 1.0000 mg/L dose group inhibited cell growth (0.071 ± 0.019, P ＜ 0.05). The apoptotic rate of 0.0500 - 1.0000 mg/L dose control group [ (18.88 ± 0.02)%,(17.58 ± 0.01)%, (17.09 ± 0.04)%, (56.00 ± 0.02)%, (57.85 ± 0.03)% ] were higher than that of the 0 mg/L group[(13.51 ± 0.01)%, all P ＜ 0.05];compared with 0 mg/L group[(25.84 ± 0.02)%], the apoptotic rate in KBD 0.0250 - 0.2500 mg/L dose group [ ( 13.69 ± 0.02) %, ( 15.96 ± 0.03 ) %, ( 16.68 ± 0.03 ) %, ( 16.67 ± 0.02) % ]were lower, and the apoptotic rate in 0.5000, 1.0000 mg/L dose group [ (59.58 ± 0.03)%, (73.48 ± 0.04)% ] were significantly higher(all P ＜ 0.05). The Fas expression in KBD 0.0500 - 0.2500 mg/L dose groups[ (41.2 ± 1.5)%,(40.3 ± 2.0)%, (50.2 ± 2.5)%] were lower than those of the same dose control group with selenium intervention [(52.4 ± 1.0)%, (67.2 ± 4.0)%, (75.1 ± 5.0)%, all P ＜ 0.05], the caspase-3 expression in KBD 0.0500,0.1000 mg/L dose groups[ (40.8 ± 1.1 )%, (45.1 ± 2.1 )%] were lower than those of the same dose control group with selenium intervention[ (68.0 ± 3.0)%, (70.6 ± 3.5)%, all P ＜ 0.05 ]. Conclusions Appropriate dose of selenium supplementation (0.1000 - 0.2500 mg/L) could promote the growth of KBD chondrocyte, decrease cell apoptosis,but have a damage when the dose of selenium ＞ 0.5000 mg/L;doses of selenium that could promote the growth of KBD chondrocyte does not mean to promote the growth of normal cartilage cells in vivo.
