【摘要】 目的观察姜黄素(curcumin,Cur)对慢性坐骨神经结扎损伤大鼠脊髓背角神经元中程序细胞死亡因子5(programmed cell death 5,PDCD5)表达的影响。方法建立慢性坐骨神经结扎损伤(chronic construction injury,CCI)模型;健康雄性SD大鼠84只,随机分为6组:空白对照组(n=4)、假手术组(Sham组)(n=16)、神经结扎组(CCI组)(n=16)和不同浓度姜黄素组[Cur 50 mg/kg组(n=16)、Cur 100 mg/kg组(n=16)及Cur 200 mg/kg组(n=16)]。Cur组行坐骨神经结扎,并腹腔注射姜黄素50、100、200 mg/(kg·d),持续至术后21 d。Sham组,CCI组和Cur组按取材时间不同分为4个亚组(n=4),即术后3 d亚组、术后7 d亚组、术后14 d亚组和术后21 d亚组。分别于术前2 d及3 d、7 d、14 d和21 d测定行为学;采用末端生脱氧核糖核苷酸转移酶介导生物素标记(terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling,TUNEL)标记法标记脊髓背角浅层凋亡细胞;免疫印迹法(Western blot,WB)法检测PDCD5蛋白的表达。结果空白组与Sham组相比,MTW和TWL差异无统计学意义(P>0.05)。CCI组术后MTW和TWL逐渐降低,在第14 d时降至最低值。给予Cur 100 mg/(kg·d)处理后,MTW和TWL逐渐增加,与CCI组在3 d、7 d、14 d及21 d比较,差异有统计学意义(P<0.05)。空白组与Sham组相比,差异无统计学意义(P>0.05)。CCI组在术后3 d即出现凋亡细胞增多,细胞凋亡高峰期出现在术后7~14 d,与Sham组相比,差异有统计学意义(P<0.05);给予不同浓度Cur处理后,凋亡细胞在术后3 d开始减少,Cur 100 mg/(kg·d)组与其他Cur组相比,差异有统计学意义(P<0.05);另可见Cur 100 mg/(kg·d)组在术后7 d、14 d及21 d,与其他Cur组、CCI组相比,差异有统计学意义(P<0.05)。空白对照组与Sham组在3 d、7 d、14 d及21 d时的PDCD5蛋白均为低水平表达,差异无统计学意义(P>0.05)。而CCI组PDCD5蛋白的表达水平,在3 d时已开始增高,在7 d及14 d时,PDCD5蛋白表达水平明显增高,14 d为PDCD5蛋白表达的高峰期,与Sham组比较,二者差异有统计学意义(P<0.05);在21 d时,PDCD5蛋白表达较7 d及14 d时降低,但仍高于3 d时的表达水平。给予低浓度Cur处理后,PDCD5蛋白表达水平较CCI组开始降低,在Cur 100 mg/(kg·d)组,PDCD5蛋白表达明显减低,与CCI组比较,二者差异有统计学意义(P<0.05)。在Cur200 mg/(kg·d)组,PDCD5蛋白表达较Cur100mg/(kg·d)组升高,二者差异有统计学意义(P<0.05)。结论姜黄素可以减轻CCI诱发的神经病理性痛行为,减少脊髓背角神经元细胞的凋亡,其机制可能与抑制PDCD5蛋白的表达有关。更多还原

【Abstract】 Objective To investigate the effects of Curcumin on expression of programmed cell death 5(PDCD5) of spinal dorsal horn in rat model of chronic construction injury(CCI) on sciatic nerve. Methods The CCI rat model was established. A total of 84 male SD rats were randomly divided into native group( n = 4),sham operation group( sham,n =16),CCI group( n = 16) and curcumin treated groups( Cur50,Cur100,Cur200,n = 16 in each group). Curcumin in the doses of 50,100,200 mg /( kg·d) was respectively given ip for 21 days after operation in group Cur50,Cur100,Cur200. Groups Sham,CCI and CUR were further divided into four subgroups according to different survival periods( 3d,7 d,14 d and 21 d). Mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL) of rats were measured on 2 days pre-operatively and 3,7,14,21days post-operatively. Apoptosis neurons in spinal superficial dorsal horn were detected using terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling( TUNEL) technique. Western blot technique was used to detect the expression of PDCD5 protein in spinal superficial dorsal horn. Results 8 There was no significant difference of MWT and TWL between sham group and native group( P > 0. 05). MWT and TWL in CCI group were significant lower than in sham group and native group( P < 0. 05) and reached the lowest at 14 d after operation. MWT and TWL in curcumin treated group( Cur100) were increased gradually and reached the highest at 14 d after operation,which were significant higher than in CCI group( P < 0. 05). There was no significant difference of apoptosis neurons between sham group and native group( P > 0. 05). Apoptosis neurons began to increase from 3 d in CCI group and reached the highest at 7 ~ 14 d after operation,which was lower than in sham group( P < 0. 05). Apoptosis neurons decreased in curcumin treated group( Cur100),which was lower than in CCI group( P < 0. 05). The level of expression of PDCD5 protein in sham group and native group was low on every post-operative day( P > 0. 05). While in CCI group,the level of expression of PDCD5 protein began to increase from 3 d and reached the highest at 7 ~ 14 d after operation,which was higher than in sham group( P < 0. 05). In curcumin treated group( Cur100),the level of expression of PDCD5 protein was significant lower than in CCI group( P < 0. 05). On the contrary,there was higher expression of PDCD5 protein in curcumin treated group( Cur200). Conclusion Curcumin could relieve the enhancement of the neuropathic pain behavior and reduce apoptosis of neurons in spinal superficial dorsal horn after the sciatie nerve ligation,which may be related with its effects of inhabiting the expression of PDCD5 protein.更多还原