目的 观察针康法对脑缺血大鼠缺血半暗区细胞凋亡及X连锁凋亡抑制蛋白(XIAP)和cleaved-caspase-9蛋白表达的影响。方法 选取180只雄性Sprague-Dawley大鼠,通过随机数字表法随机分为假手术组、模型组、针刺组、康复组和针康组。每组再分为3 d、7 d和14 d三个亚组(n=12)。采用改良Longa线栓法进行造模,假手术组和模型组不予治疗,针刺组进行头穴丛刺治疗,康复组进行跑台康复训练,针康组进行针康法治疗。术后各时间点对大鼠进行改良神经功能缺损评分(mNSS)后取材,采用TUNEL染色观察大鼠脑缺血半暗区细胞凋亡率,Western blotting检测脑缺血半暗区XIAP和cleaved-caspase-9蛋白表达。结果 术后各时间点,与模型组比较,各治疗组mNSS评分均降低(P<0.05),细胞凋亡率均降低(P<0.05),XIAP蛋白表达上调(P<0.05),cleaved-caspase-9蛋白表达下调(P<0.05);与针刺组、康复组比较,针康组细胞凋亡率进一步降低(P<0.05),XIAP蛋白表达进一步上调(P<0.05)。术后7 d、14 d,针康组mNSS评分进一步降低(P<0.05),针康组cleaved-caspase-9蛋白表达进一步下调(P<0.05)。结论 针康法能降低脑缺血大鼠神经功能缺损,优于单独的头穴丛刺和康复训练,其机制可能与促进XIAP蛋白表达,抑制caspase-9蛋白活化,从而减少细胞凋亡有关。
唐强
,
田源
,
叶涛
,
朱路文
,
阮野
,
赵彬
,
陈慧杰
,
王雪
. 针康法对脑缺血大鼠缺血半暗区细胞凋亡及X连锁凋亡抑制蛋白和cleaved-caspase-9蛋白表达的影响[J]. 中国康复理论与实践, 2017
, 23(12)
: 1365
-1371
.
DOI: 10.3969/j.issn.1006-9771.2017.12.001
Objective To explore the effect of acupuncture-rehabilitation therapy on apoptosis and the expression of X-linked inhibitor of apoptosis protein (XIAP) and cleaved-caspase-9 in ischemic penumbra of rats with cerebral ischemia. Methods A total of 180 male Sprague-Dawley rats were randomly divided into sham operation group, model group, acupuncture group, rehabilitation group and acupuncture-rehabilitation therapy group. Each group was divided into three days, seven days and 14 days subgroups (n=12). The model was established with the modified Longa suture method. The model group and the sham operation group received no treatment. The acupuncture group received cluster needling of scalp acupuncture, the rehabilitation group received treadmill training, and the acupuncture-rehabilitation therapy group received both acupuncture and treadmill training. On the third, seventh and 14th days after modeling, the neurological function was assessed with modified Neurological Severity Score (mNSS). The apoptosis was observed by TUNEL staining and the expression of XIAP and cleaved-caspase-9 protein was determined by Western blotting in cerebral ischemic penumbra, respectively. Results Compared with the model group, the mNSS scores decreased, the rate of apoptosis decreased, the expression of XIAP protein increased, and the expression of cleaved-caspase-9 decreased in each treatment group at each time point (P<0.05). Compared with the other two treatment groups, the rate of apoptosis further decreased at each time points (P<0.05), the expression of XIAP protein further increased at each time points (P<0.05), the mNSS score further decreased (P<0.05) and the expression of cleaved-caspase-9 protein further decreased (P<0.05) seven days and 14 days after modeling in the acupuncture-rehabilitation therapy group. Conclusion Acupuncture and rehabilitation therapy could reduce the neurological deficit in rats with cerebral ischemia, which was superior to the simple acupuncture treatment and rehabilitation training. The mechanism may be related to th reduction of apoptosis by promoting XIAP protein expression and inhibiting caspase-9 protein activation.
[1] 魏琰,张维文,王晓莉,等. 无症状性脑梗死患者首发症状性急性脑卒中的临床特征及相关危险因素[J]. 脑与神经疾病杂志, 2016, 24(11): 681-684.
[2] 关莹,张立,邢艳丽,等. 针康法对脑卒中后痉挛状态的影响[J]. 中国康复理论与实践, 2011, 17(4): 325-327.
[3] 唐强,吴北峰,朱肖菊. 针康法对缺血性脑卒中患者肢体运动功能的影响[J]. 针灸临床杂志, 2005, 21(2): 46-47.
[4] 唐强,李宏玉,朱路文,等. 针康法对脑卒中后非流畅性失语患者言语功能的影响[J]. 针灸临床杂志, 2016, 32(7): 35-37.
[5] 唐强,朱路文. 脑卒中康复新策略&#x#x02014;&#x#x02014;针康法[J]. 中国康复医学杂志, 2015, 30(10): 1071-1073.
[6] Russell JC, Whiting H, Szuflita N, et al. Nuclear translocation of X-linked inhibitor of apoptosis (XIAP) determines cell fate after hypoxia ischemia in neonatal brain [J]. J Neurochem, 2008, 106(3): 1357-1370.
[7] Shiozaki EN, Chai J, Rigotti DJ, et al. Mechanism of XIAP-mediated inhibition of caspase-9 [J]. Mol Cell, 2003, 11(2): 519-527.
[8] Zumbr&#x#x000e4;gel FK, Machtens DA, Curth U, et al. Survivin does not influence the anti-apoptotic action of XIAP on caspase-9 [J]. Biochem Biophys Res Commun, 2016, 482(4): 530-535.
[9] Yang LX, Zhang X, Zhao G. Ginsenoside Rd attenuates DNA damage by increasing expression of DNA glycosylase endonuclease VIII-like proteins after focal cerebral ischemia [J]. Chin Med J (Engl), 2016, 129(16): 1955-1962.
[10] 唐强,叶涛,朱路文,等. 针康法对脑缺血大鼠神经功能和细胞外信号调节激酶1/2信号通路的影响[J]. 中国康复理论与实践, 2017, 23(1): 27-31.
[11] 唐强,姜云飞,朱路文,等. 针康法对局灶性脑缺血大鼠神经功能缺损及缺血区碱性成纤维细胞生长因子、血管抑制素蛋白表达的影响[J]. 中国康复理论与实践, 2015, 21(10): 1151-1155.
[12] 孔妍. 针康法对局灶性脑缺血大鼠胶质细胞环境和细胞凋亡的影响[D]. 哈尔滨:黑龙江中医药大学, 2013.
[13] 叶涛,唐强,朱路文,等. 针康法对脑缺血大鼠神经功能和缺血半暗区皮层血管内皮生长因子受体表达的影响[J]. 中国康复理论与实践, 2017, 23(5): 520-524.
[14] 崔乃松. 针康法对脑缺血大鼠运动功能及缺血区皮质Nogo-A表达的影响[D]. 哈尔滨:黑龙江中医药大学, 2013.
[15] 陈慧杰,唐强,朱路文,等. 针康法对脑缺血大鼠脑组织微血管内皮超微结构及aFGF、PF4蛋白表达影响[J]. 针灸临床杂志, 2017, 33(5): 65-69.
[16] Zhang X, Tang N, Hadden TJ, et al. Akt, FoxO and regulation of apoptosis [J]. Biochim Biophys Acta, 2011, 1813(11): 1978-1986.
[17] Hermeking H. The miR-34 family in cancer and apoptosis [J]. Cell Death Differ, 2010, 17(2): 193-199.
[18] 赵捷,王静,吴精霞. 五价钒致神经细胞凋亡中细胞色素C和天冬氨酸特异性的半胱氨酸蛋白水解酶9及3蛋白的变化[C]. 2015中国金属学会冶金安全与健康分会预防医学专业委员会学术交流会, 2015.
[19] 彭伟,骆泓洁,元小冬. 死亡受体介导的细胞凋亡研究进展[J]. 生命的化学, 2016, 36(5): 629-632.
[20] 刘楠,李岩,牛振华,等. 内质网应激介导的神经细胞凋亡通路[J]. 中风与神经疾病, 2013, 30(8): 757-759.
[21] 陈琳,吴巧凤,杨明晓,等. 基于细胞凋亡探讨针刺作用机理的研究述评[J]. 上海针灸杂志, 2016, 35(10): 1143-1146.
[22] Huang C, Zeng X, Jiang G, et al. XIAP BIR domain suppresses miR-200a expression and subsequently promotes EGFR protein translation and anchorage-independent growth of bladder cancer cell [J]. J Hematol Oncol, 2017, 10(1): 6.
[23] Holcik M, Gibson H, Korneluk RG. XIAP: apoptotic brake and promising therapeutic target [J]. Apoptosis, 2001, 6(4): 253-261.
[24] Ch&#x#x000e1;vez-Gal&#x#x000e1;n L, Sada-Ovalle I, Baez-Salda&#x#x000f1;a R, et al. Monocytes from tuberculosis patients that exhibit cleaved caspase 9 and denaturalized cytochrome C are more susceptible to death mediated by Toll-like receptor 2 [J]. Immunology, 2012, 135(4): 299-311.
[25] 曾芳,何宇恒,彭静,等. 电针对SAMP8小鼠海马神经元凋亡蛋白Caspase-3、Caspase-9及凋亡抑制蛋白XIAP表达的影响[J]. 中国老年学, 2008, 28(6): 521-523.
[26] 赵雯,马艳红,韩咏竹,等. 肝豆汤改良方对TX小鼠脑组织XIAP、Caspase-9和Caspase-3蛋白及其mRNA表达的影响[J]. 安徽中医药大学学报, 2015, 34(4): 58-64.
