糖尿病大鼠椎间盘组织中戊糖甙素与细胞凋亡的关系(英文)

Relationship between pentosidine and cellular apoptosis in the intervertebral disc of diabetic rats

背景细胞凋亡可能是引起椎间盘细胞减少的一个主要因素。但是关于诱导椎间盘细胞凋亡的生物学因素还不清楚。目的通过观察糖尿病大鼠椎间盘细胞凋亡情况,探讨可能与凋亡有关的因素。设计随机对照的实验研究。地点和对象实验地点在上海九院实验动物中心和上海第二医科大学生物物理教研室完成。36只雄性SD大鼠(4月龄),购自中科院上海实验动物中心,平均体质量244g。按随机数字表法分为2组,每组18只。干预措施处理组用腹腔单次注射链脲佐菌素溶液(streptozotocin,STZ,40mg/kg)造成糖尿病。正常对照组,仅给予等量的柠檬酸缓冲液。72h后测定大鼠的血糖,血糖在16.0mmol/L以上者确定为糖尿病。分别在第1,3,4个月时每组各取6只大鼠处死,取其腰椎间盘组织,进行测定。主要观察指标用流式细胞术检测椎间盘细胞凋亡情况。用高效液相层析法测定椎间盘组织中戊糖甙素含量变化。结果处理组大鼠在注射STZ溶液后,血糖水平均高于16.0mmol/L[(23.71±2.69)mmol/L]。在不同时期,糖尿病大鼠组椎间盘细胞凋亡率(%)分别是18.52±4.81,18.74±5.13,19.41±5.22,明显高于同期对照组4.59±2.24,9.38±2.37,10.78±3.86(P<0.05),糖尿病大鼠椎间盘组织中戊糖甙素含量(nmol/g)分别是33.53±5.88,37.03±12.77,36.53±11.95,明显高于同?

BACKGROUND:Apoptosis has been suggested to be the main factor for the loss of the cells in the intervertebral disc. But the biological causes of disc apoptosis are unclear. OBJECTIVE:To examine the apoptosis of the intervertebral disc cells of diabetic rats for exploring the possible factor related to the apoptosis.DESIGN:A randomized controlled experimental study.SETTING and PARTICIPANTS:Thirty six male SD rats of 4 months were randomly put into two groups with random number table method,and 18 rats in each group.The study was conducted in the Laboratory Animal Center of Shanghai Ninth Hospital and the Biophysics Department of Shanghai Second Medical University.INTERVENTION:The diabetes mellitus(DM)was induced by a single intraperitoneal injection of streptozotocin(STZ) solution(40 mg/kg).In control group,the rats were injected with the sodium citrate buffer of the same volume.The blood glucose level was measured 72 hours later.The standard for the DM is above 16.0 mmol/L.The rats were killed 1 month,3 months and 4 months later respectively.Each time 6 diabetic rats and 6 control rats were sacrificed respectively with the lumbar intervertebral discs for measurement.MAIN OUTCOME MEASURES:The apoptotic rate was measured with flow cytometry.The measurement of the pentosidine was performed with high performance liquid chromatography.RESULTS:The venous full blood glucose levels of all the STZ induced rats were above 16.0 mmol/L[(23.71±2.69) mmol/L],which were higher significantly than those of the control rats[(4.19±0.34)mmol/L](P< 0.01).At three time points,the percentages of apoptotic intervertebral disc cells in the diabetic rats were 18.52±4.81,18.74±5.13,19.41±5.22. They were 4.59±2.24,9.38±2.37,10.78±3.86 in control rats.There were significantly differences between the two groups(P< 0.05).The levels(nmol/g)of pentosidine in the intervertebral discs of the diabetic rats were 33.53±5.88,37.03±12.77,36.53±11.95.They were 13.87±3.30,13.60±2.56,15.33±4.04 in control rats. There were significantly differences between the two groups(P< 0.05).There was a positive correlation between the levels of pentosidine and the percentage of apoptotic intervertebral cells of all groups respectively(Pearson correlation coefficient:r=0.857-0.970,P< 0.05).CONCLUSION:The apoptosis of the intervertebral disc cells in diabetic rats increased significantly.Pentosidine and other advanced glycation end products may be related to the apoptosis of the intervertebral disc cells in rats.