[电针通过下调脊髓损伤大鼠细胞磷脂酶 A2 和减少神经细胞凋亡改善肢体运动功能]

Hai-Hua Yao, You-Jiang Min, Dong-Ying Hong, Li Wang, Xiu-Yun Lu, Yi-Hua Yang
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摘要

目的观察电针(EA)对细胞膜磷脂酶 A2(cPLA2)表达的影响。对脊髓损伤(SCI)大鼠细胞膜磷脂酶 A2(cPLA2)表达和神经细胞凋亡的影响。方法:将72只雌性SD大鼠随机分为模型组、EA组、拮抗剂组和EA+拮抗剂组,每组18只,其余18只作为假手术(sham)组。组。SCI模型参照改良Allen法,用重量冲击器建立。后肢运动功能采用巴索-巴蒂-布雷斯纳汉(BBB)评分。EA组大鼠分别在 "大水"(GV14)、"窑阳关"(GV3)、双侧 "敕勒"(BL32)和 "祖山里"(BL32)处接受EA刺激。和 "足三里"(ST36)。20分钟,每天一次,共14天。拮抗剂组大鼠先接受静脉注射,然后腹腔注射花生四烯丙基三氟甲基酮(AACOCF3)(cPLA2 的拮抗剂),隔日一次。EA+拮抗剂组大鼠在接受EA治疗的同时注射拮抗剂。治疗后,大鼠被处死,收集脊髓组织,采用Western blot检测cPLA2、p-cPLA2、Bcl-2、Bax和Caspase-3的蛋白表达,采用qRT-PCR检测cPLA2、Bcl-2、Bax和Caspase-3的mRNA表达。Nissl染色法检测脊髓的形态学变化:结果:与假组相比,BBB评分、Bcl-2蛋白和mRNA的表达均明显下调(PPPPPPP结论:EA可改善小鼠的肢体运动功能:EA可改善SCI大鼠的肢体运动功能,这可能与其下调p-cPLA2、Bax和Caspase-3的表达,上调Bcl-2以减少脊髓区域神经细胞凋亡的功能有关。
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[Electroacupuncture improves limb motor function by down-regulating cytosolic phospholi-pase A2 and reducing apoptosis of nerve cells in rats with spinal cord injury].

Objective: To observe the effect of electroacupuncture(EA) on the expression of cytosolic phospholipase A2 (cPLA2) and apoptosis of nerve cells in rats with spinal cord injury (SCI), so as to explore its mechanisms underlying improvement of SCI.

Methods: Seventy-two female SD rats were randomly divided into model, EA, antagonist and EA+antagonist groups, with 18 rats in each group and other 18 rats were used as the sham operation (sham) group. The SCI model was established by referring to modified Allen's method with a weight impactor. The hindlimb motor function was assessed by using Basso-Beattie-Bresnahan (BBB) score. Rats of the EA group were subjected to EA stimulation at "Dazhui"(GV14), "Yaoyangguan"(GV3), bilateral "Ciliao"(BL32) and "Zusanli"(ST36) for 20 min, once a day for 14 days. Rats of the antagonist group received intravenous injection followed by intraperitoneal injection of arachidonyl trifluoromethyl ketone (AACOCF3, antagonist of cPLA2), once every other day. Rats of the EA+antagonist group received EA treatment combined with antagonist injection. After the treatment, the rats were sacrificed and the spinal cord tissue was collected for detecting the protein expression of cPLA2, p-cPLA2, Bcl-2, Bax and Caspase-3 by Western blot, and the mRNA expression of cPLA2, Bcl-2, Bax and Caspase-3 using qRT-PCR. The morphological changes of the spinal cord were detected by Nissl staining.

Results: In comparison with the sham group, the BBB score, expression of Bcl-2 protein and mRNA were significantly down-regulated (P<0.01), whereas the expression levels of Bax, Caspase-3 and p-cPLA2 proteins and mRNAs were considerably up-regulated in the model group (P<0.01). Compared with the model group, the BBB score, expression levels of Bcl-2 protein and mRNA were significantly up-regulated (P<0.01, P<0.05), while the expression levels of Bax, Caspase-3 and p-cPLA2 proteins in the EA, antagonist and EA+antagonist groups, Bax and cPLA2 mRNAs in both antagonist and EA+antagonist groups, and Caspase-3 mRNA in the EA+antagonist group were obviously down-regulated (P<0.01, P<0.05). The effect of EA+antagonist was significantly superior to EA in increasing BBB score and in lowering expression of Bax and cPLA2 mRNAs (P<0.01, P<0.05). Nissl staining showed reduced number of nerve cells and Nissl bodies, and striped dark blue cells in the model group, which was milder in the EA and antagonist groups, particularly in the EA+antagonist group.

Conclusion: EA may improve the limb motor function of SCI rats, which may be related to its functions in down-regulating the expression of p-cPLA2, Bax and Caspase-3 and up-regulating Bcl-2 to reduce the apoptosis of nerve cells in the regional spinal cord.

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