地塞米松诱导的危重病性肌病大鼠骨骼肌Beclin1、LC3的表达
Expression of Beclin1 and LC3 in dexamethasone-induced rat critical illness myopathy
目的 观察地塞米松对大鼠危重病性肌病（CIM）比目鱼肌细胞自噬相关因子Beclin1和LC3表达的影响，探讨地塞米松诱导的大鼠危重病性肌病的可能机制。 方法 将健康SD大鼠分为对照组和实验组；实验组又分为7,9,11d 3个时相点（n=10）。实验组采用5mg/kg地塞米松连续腹腔注射，每天1次，对照组腹腔注射等量的生理盐水。采用足迹法判定肌功能缺损情况。利用免疫组化和Western blot检测比目鱼肌细胞自噬相关因子Beclin1和LC3的表达情况。 结果 与对照组相比，实验组大鼠出现不同程度肌肉功能缺损症状，以11d时大鼠缺损程度最为严重。Western blot结果显示，对照组或可见微弱的Beclin1、LC3阳性表达，随着时间延长，实验组可见明显的Beclin1、LC3阳性表达，以11d时相点表达最为明显。免疫组化结果也证实了同一趋势。 结论 地塞米松诱导的大鼠CIM可能通过激活Beclin1和LC3的表达从而发挥对细胞自噬的调节作用。
Objective To investigate the expression of critical illness myopathy on autophagy-related factors Beclin1 and LC3 in rat skeletal muscle. Method SD rats were divided into control and experimental groups; Experimental groups were divided into 3 time points, i.e., 7, 9 and 11d（n=10）.Experimental groups underwent intraperitoneal injection of dexamethasone 5 mg/kg once per day, and the healthy control group was injected with normal saline. The muscle function defect was determined by footprint method. The expression of skeletal muscle autophagy-related factors Beclin1 and LC3 was examined by immunohistochemistry and western blot analysis. Results Compared with the control group, rats in the experimental groupdemonstrated muscle function impairment symptoms of varying degrees ,which was most serious at 11d time point. Western blot results showed that Beclin1 and LC3 positive expression in the control group, or Beclin1 and LC3 expression were significantly higher in the experimental group than that in the control group. Immunohistochemistry confirmed the same trend. Conclusion Beclin1 and LC3 may play an important role in cell autophagy of skeletal muscle in critical illness myopathy in rats.
[1] Glance LG, Li Y, Osler TM, et al. Impact of patient volume on the mortality rate of adult intensive care unit patients[J]. Crit Care Med, 2006, 34(7):1925-1934.
[2] de Jonghe B, Lacherade JC, Sharshar T, et al. Intensive care unit-acquired weakness: risk factors and prevention[J]. Crit Care Med, 2009, 37(10): S309-315.
[3] Weber-Carstens S, Deja M, Koch S, et al. Risk factors in critical illness myopathy during the early course of critical illness: a prospective observational study[J]. Crit Care, 2010,14(3):R119-120.
[4] Mariño G, López-Otín C. Autophagy: molecular mechanisms, physiological functions and relevance in human pathology [J]. Cell Mol Life Sci, 2004, 61(12): 1439-1454.
[5] 秦正红, 乐卫东. 自噬-生物学与疾病[M]. 北京:科学出版社, 2011:182-199.
[6] Tsuchihara K, Fujii S, Esumi H. Autophagy and cancer: Dynamism of the metabolism of tumor cells and tissues[J]. Cancer Lett, 2009, 278(2): 130-138.
[7] 袁静, 梁佳, 赵颂, 等. 地塞米松诱导的危重病性肌病的细胞死亡方式[J]. 第三军医大学学报, 2012, 34(5):453-455.
[8] Chen N, Karantza-Wadsworth V. Role and regulation of autophagy in cancer[J]. Biochim Biophys Acta, 2009, 1793(9): 1516-1523.
[9] Kang R, Zeh HJ, Lotze MT, et al. The beclin-1 network regulates autophagy and apoptosis[J]. Cell Death Differ, 2011, 18(4): 571-580.
[10]Deretic V, Saitoh T, Akira S. Autophagy in infection, inflammation and immunity[J]. Nat Rev Immunol, 2013,13(10): 722-737.
[11]Baerga R, S Jin, M Kulkum, et al. Dentification and characterization of Beclin1 interacting proteins and Beclin1 post-translational modifications[J]. AACR, 2005,2005(1): 586-587．
[12]Yue ZY, Zhong Y. From a global view to focused examination: undersdanding cellular function of lipid kinase VPS34-Beclin-1 complex inautophagy[J]. J Mol Cell Biol, 2010, 2(6): 305-307.
[13]宋丽萍, 李翠兰, 王子介, 等. 自噬与卵巢癌研究进展[J]. 中华临床医师杂志, 2015, 9(3):464- 467.
[14]Wirth M, Joachim J, Tooze SA. Autophagosome formation the role of ULK1 and Beclin1-PI3KC3 complexes in setting the stage[J]. Semin Cancer Biol, 2013, 23(5): 301-309.
[15]沈扬, 梁立治, 洪明晃, 等. 微管相关蛋白LC3和自噬基因Beclin1在上皮性卵巢癌中的表达及其意义[J]. 癌症, 2008, 27(6):595-599.
[16]Mizushima N. Methods for monitoring autophagy[J]. Int J Biochem Cell Biol, 2004, 36(12): 2491-2502.
[17]Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing[J]. EMBO J, 2000, 19(21): 5720-5728.
[18]Yewdell JW, Antón LC, Bennink JR. Defective ribosomal products (DRiPs): a major source of antigenic peptides for MHC classⅠmolecules [J]? J Immunol, 1996, 157(5): 1823-1826.
[19] Myeku N, Figueiredo-Pereira ME. Dynamics of the degradation of ubiquitinated proteins by proteasomes and autophagy: association with sequestosome 1/p62 [J]. J Biol Chem, 2011, 286(25): 22426-22440.
[20]张创杰,张连峰,周琳.自噬相关蛋白Beclin1、LC3和P62在进展期胰腺癌中的表达及临床意义[J].世界华人消化杂志,2015,23(2):318-323.
[21]靳航.自噬与泛素-蛋白酶体系统在实验性脑出血灶周组织损伤中作用的研究[D]. 吉林：吉林大学, 2013.
[22] Tung YT, Hsu WM, Lee H, et al. The evolutionarily conserved interaction between LC3 and p62 selectively mediates autophagy-dependent degradation of mutant huntingtin[J]. Cell Mol Neurobiol, 2010, 30(5):795-806.
[23]屈惠莹,包翠芬,于迪,等.诱导的危重病性肌病大鼠骨骼肌p62和泛素的表达[J].中国临床解剖学杂志, 2014,32(5):599-603.
国家自然科学基金（31170930）；
京ICP备08002354号-3/
| 〈 |
|
〉 |
