北部战区总医院骨科,辽宁 沈阳 110001
Published:25 January 2025,
Received:26 September 2023,
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WANG LING, WANG SHUANG, LIU YING, et al. Platelet lysate improving mitochondrial function of astrocytes after spinal cord injury. [J]. Journal of regional anatomy and operative surgery, 2025, 34(1): 16-20.
WANG LING, WANG SHUANG, LIU YING, et al. Platelet lysate improving mitochondrial function of astrocytes after spinal cord injury. [J]. Journal of regional anatomy and operative surgery, 2025, 34(1): 16-20. DOI: 10.11659/jjssx.09E023114.
目的
2
探讨血小板裂解液对脊髓损伤星形胶质细胞线粒体功能的影响。
方法
2
在星形胶质瘤细胞SVGp12中使用不同浓度H
2
O
2
(100 µmol/L、400 µmol/L、1 000 µmol/L、2 000 µmol/L)孵育,CCK-8检测OD值,选取细胞抑制率约50%的H
2
O
2
浓度进行后续实验。使用不同浓度血小板裂解液(血小板裂解液与培养基体积比分别为1∶10、1∶40、1∶80、1∶160、1∶320)孵育细胞24 h,测定OD值,根据细胞存活率筛选最佳血小板裂解液浓度。将SVGp12细胞分为正常组(正常培养,无特殊处理)、模型组(加入1 000 μmol/L H
2
O
2
孵育3 h)、治疗组(加入1 000 μmol/L H
2
O
2
孵育3 h后,加入体积比为1∶80的血小板裂解液处理),采用线粒体活性染色、线粒体膜电位检测、线粒体通透性转换孔评估线粒体功能。
结果
2
1 000 μmol/L H
2
O
2
孵育细胞3 h后,细胞抑制率为48%,OD值显著降低(
P
<
0.01),证实星形胶质细胞损伤模型成功建立。经不同浓度血小板裂解液处理后细胞OD值及细胞存活率均高于模型组(
P<
0.05),选取细胞存活率为20%的血小板裂解液浓度(体积比1∶80)进行后续实验。血小板裂解液能改善损伤后星形胶质细胞的形态。治疗组细胞线粒体活性显著高于模型组,模型组细胞线粒体活性显著低于正常组,差异均有统计学意义(
P
<
0.05)。模型组线粒体膜电位较正常组减少,治疗组线粒体膜电位较模型组增加,差异均有统计学意义(
P
<
0.01)。模型组线粒体膜通透性较正常组增加,治疗组线粒体膜通透性较模型组降低,差异均具有统计学意义(
P
<
0.01)。
结论
2
血小板裂解液能够提高脊髓损伤星形胶质细胞的存活率,增强细胞线粒体活性,改善线粒体膜电位和线粒体通透性转换孔开放情况。
Objective
2
To investigate the effect of platelet lysate on mitochondrial function of astrocytes after spinal cord injury.
Methods
2
Astrocytoma cells SVGp12 were incubated with different concentrations
of H
2
O
2
(100 µmol/L,400 µmol/L,1 000 µmol/L,2 000 µmol/L),and the OD value was determined by CCK-8 assay. The H
2
O
2
concentration with a cell inhibition rate of about 50% was selected for follow-up experiments. The cells were incubated with different concentrations of platelet lysate (the volume ratios of platelet lysate to culture medium were 1∶10,1∶40,1∶80,1∶160 and 1∶320) for 24 hours,and the OD value was determined,thereby selecting the optimal concentration of platelet lysate based on the cell survival rate. SVGp12 cells were divided into the normal group (cultured normally without special treatment),the model group (incubated with 1 000 µmol/L of H
2
O
2
for 3 hours),and the treatment group (incubated with 1 000 µmol/L of H
2
O
2
for 3 hours and then treated with platelet lysate at a volume ratio of 1∶80). The mitochondrial function was evaluated by mitochondrial activity staining,mitochondrial membrane potential detection,and mitochondrial permeability transition pore detection.
Results
2
After incubating cells with 1 000 µmol/L of H
2
O
2
for 3 hours,the cell inhibition rate was 48%,and the OD value decreased significantly (
P
<
0.01),confirming the successful establishment of a model of astrocytes with spinal cord injury. After treated with different concentrations of platelet lysate,the OD values and cell survival rate of the cells were higher than those in the model group (
P
<
0.05). The concentration of platelet lysate (volume ratio of 1∶80) with cell viability of 20% was selected for follow-up experiments. Platelet lysate could improve the morphology of injured astrocytes. The mitochondrial activity of the treatment group was significantly higher than that of the model group,and the mitochondrial activity of the model group was significantly lower than that of the normal group,with statistically significant differences (
P
<
0.05). The mitochondrial membrane potential in the model group was lower than that in the normal group,and the mitochondrial membrane potential in the treatment group was higher than that in the model group,with statistically significant differences (
P
<
0.01). The mitochondrial permeability in the model group was greater than that in the normal group,and the mitochondrial permeability in the treatment group was smaller than that in the model group,with statistically significant differences (
P
<
0.01).
Conclusion
2
Platelet lysate can improve the survival rate of astrocytes after spinal cord injury,enhance the mitochondrial activity of cells,and improve the mitochondrial membrane potential and mitochondrial permeability transition pore opening.
血小板裂解液脊髓损伤星形胶质细胞线粒体活性线粒体膜电位线粒体通透性转换孔
platelet lysatespinal cord injuryastrocytesmitochondrial activitymitochondrial membrane potentialmitochondrial permeability transition pore
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