目的探讨在增龄过程中SD大鼠髓核组织中Beclin-1和微管相关蛋白轻链3(MAPLC3)的表达及其意义。方法3个月龄、12个月龄、24个月龄SD大鼠各8只,建立青年组、中年组和老龄组大鼠模型,苏木素-伊红(HE)染色检测椎间盘组织的退变;透射电镜检测髓核细胞中自噬体的表达;免疫组织化学染色和逆转录-聚合酶链反应(RT-PCR)法测定髓核组织中Beclin-1和MAPLC3的表达。结果大鼠增龄过程可较好地体现椎间盘的退变变化;不同年龄组大鼠髓核细胞中均有自噬体的表达;Beclin.1和MAPLC3在不同年龄组sD大鼠髓核细胞中均有表达(0.42±0.05,0.47±0.06,0.52±0.05;0.34±0.06,0.39±0.05,0.46±0.09),且老龄组表达均较青年组高(P〈0.01)。Beclin-1和MAPLC3mRNA在不同年龄组SD大鼠髓核组织中均有表达(0.86±0.12,0.93±0.14,1.01±0.13;1.09±0.06,1.15±0.07,1.33±0.11),且老龄组表达较青年组表达明显增高(分别为P〈0.05和P〈0.01)。结论自噬存在于椎间盘退变过程中,且MAPLC3与Beclin-1在椎间盘退变过程中的表达增加,提示自噬活性的增加可能与椎间盘退变的发展进程有关。
Objective To evaluate the expression and significance of Beclin-1 and microtubule-as- sociated protein 1 light chain 3 ( MAPLC3 ) in nucleus pulpous of rats during the aging process. Methods The models of youth, middle-aged and old rats were made with the 3-month, 12-month and 24-months SD rats respectively. The grades of intervertebral disc degeneration were measured with HE staining. The ex- pression of autophagosome in nucleus pulpous was detected under a transmission electron microscope (TEM). The changes of Beclin-1 and MAPLC3 in nucleus pulpous were assayed by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results The aging process of rats may reflect the process of intervertebral disc degeneration. There was expression of autophagosome in nucleus pulpous in every group. Beclin-1 and MAPLC3 were expressed in nucleus pulpous cells in each group ( average absorbance values were 0. 42 ± 0. 05, 0. 47 ± 0. 06, 0. 52 ± 0. 05 and 0. 34 ± 0. 06, 0. 39 ± 0. 05, 0.46 ± 0. 09 respectively) and both of them were higher in old group (P 〈 0. 01 ). Bcclin-1 and MAPLC3 mRNA was expressed in nucleus pulpous in each group (0. 86 ± 0. 12, 0. 93 ± 0. 14, 1.01 ± 0. 13 and 1. 09 ± 0. 06 ,1.15 ± 0. 07, 1.33 ±0. 11 respectively) and both of them were higher in old group (P 〈 0. 05 and P 〈 0.01 respectively). Conclusion Autophagy resides in the process of intervertebral disc degeneration. The expression of MAPLC3 and Beclinl was up-regulated in nucleus pulpous in different groups and autophagy. Up-regulation of MAPLC3 and Beclinl may be related to the process of the lumbar disc degeneration.