论文部分内容阅读
目的研究NF-κB抑制剂吡咯烷二硫代氨基甲酸酯(pyrrolidine dithiocarbamate,PDTC)延缓失神经肌萎缩的效果及其机制。方法健康成年Wistar大鼠30只,雌雄不限,体重(200±10)g。实验动物随机分为3组,即正常对照组(A组,n=6)、失神经对照组(B组,n=12)和PDTC治疗组(C组,n=12)。A组仅暴露右侧坐骨神经并不切断;B、C组切断右侧坐骨神经0.7～1.0 cm,建立右下肢腓肠肌失神经支配模型后,C组采用PDTC 100 mg/(kg?d)腹腔注射,B组以等量生理盐水腹腔注射。A组术后当天和B、C组术后14、28 d,完整分离双侧腓肠肌检测肌肉湿重维持率,采用Western blot检测腓肠肌NF-κB p65表达水平,激光共聚焦扫描显微镜观察线粒体通透性转换孔(mitochondrial permeability transition pore,MPTP)开放情况,TUNEL法检测细胞凋亡情况。结果 A组腓肠肌湿重维持率为1.039±0.115;与A组比较,B、C组各时间点腓肠肌湿重维持率均明显下降,且C组腓肠肌湿重维持率明显高于同期B组,差异均有统计学意义(P<0.05)。A组NF-κB p65表达为0.224±0.041;与A组比较,B、C组各时间点NF-κB p65表达明显升高,且C组NF-κB p65表达明显低于同期B组,差异均有统计学意义(P<0.05)。A组MPTP荧光强度为31.582±1.754;与A组比较,B、C组各时间点MPTP荧光强度均降低,且C组MPTP荧光强度明显高于同期B组,差异均有统计学意义(P<0.05)。A组有少量细胞凋亡,细胞凋亡率为4.542%±0.722%;与A组比较,B、C组各时间点细胞凋亡率均明显增加,且C组细胞凋亡率明显低于同期B组,差异均有统计学意义(P<0.05)。结论 PDTC可有效延缓失神经肌萎缩,其作用机制与抑制NF-κB表达、降低骨骼肌细胞MPTP开放及抑制骨骼肌细胞凋亡有关。
Objective To investigate the effect and mechanism of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, on denervation and amyotrophy in rats. Methods Thirty healthy adult Wistar rats were male and female, weighing 200 ± 10 g. The experimental animals were randomly divided into three groups: normal control group (n = 6), denervation control group (n = 12) and PDTC treatment group (n = 12). In group A and B, the right sciatic nerve was not cut off. In group B and C, the right sciatic nerve was cut off by 0.7-1.0 cm. After establishing the denervated model of right lower extremity gastrocnemius muscle, group C received intraperitoneal injection of PDTC 100 mg / (kg? D) Group with the same amount of saline intraperitoneal injection. Muscle wet weight maintenance rate was measured on the postoperative day and in group B and C at 14 and 28 days after operation. The expression of NF-κB p65 in gastrocnemius muscle was detected by Western blot. The mitochondrial permeability was observed by laser scanning confocal microscopy The opening of mitochondrial permeability transition pore (MPTP) and the apoptosis were detected by TUNEL method. Results The maintenance rate of gastrocnemius wet weight in group A was 1.039 ± 0.115. Compared with group A, the maintenance rate of gastrocnemius wet weight in group B and C were significantly decreased, and the maintenance rate of gastrocnemius wet weight in group C was significantly higher than that in group B All were statistically significant (P <0.05). The expression of NF-κB p65 in group A was 0.224 ± 0.041. Compared with group A, the expression of NF-κB p65 in group B and C were significantly increased at each time point, and the expression of NF-κB p65 in group C was significantly lower than that in group B There was statistical significance (P <0.05). The MPTP fluorescence intensity of group A was 31.582 ± 1.754. Compared with group A, the MPTP fluorescence intensity of group B and C decreased at each time point, and the fluorescence intensity of MPTP in group C was significantly higher than that of group B at the same time (P < 0.05). In group A, there was a small amount of apoptosis with a rate of 4.542% ± 0.722%. Compared with group A, the rate of apoptosis was significantly increased at each time point in group B and C, and the apoptosis rate in group C was significantly lower than that in the same period B group, the difference was statistically significant (P <0.05). Conclusions PDTC can effectively delay the development of denervation and atrophy, and its mechanism is related to inhibiting the expression of NF-κB, decreasing the opening of MPTP in skeletal muscle cells and inhibiting the apoptosis of skeletal muscle cells.