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目的:探讨脊髓损伤(SCI)后丹参注射液促进大鼠脊髓神经功能恢复的机制。方法:大鼠(清洁级)随机分为正常组、模型组、给药组、信号通路阻断组和空白组(n=10)。除正常组外,其余各组大鼠均建立重力打击SCI模型。模型组伤后常规饲养;给药组伤后1 h腹腔注射丹参注射液(1 m L·kg~(-1));信号通路阻断组伤后1 h腹腔注射丹参注射液(1 m L·kg~(-1),含雷帕霉素3 mg·kg~(-1));空白组伤后给予腹腔注射生理盐水(1 m L·kg~(-1))。上述各组均按1次/d注射。14 d后处死动物,处死前采用联合行为评分法(CBS)评价大鼠脊髓神经功能恢复情况,并取损伤节段上下1 cm的脊髓组织。分别采用免疫组化SP法,蛋白质免疫印迹(Western blot)和逆转录-聚合酶链式反应(RT-PCR)技术检测各实验组大鼠PI3K,Akt和m TOR蛋白及mRNA的表达水平。结果:与模型组、信号通路阻断组和空白组大鼠比较,给药组大鼠CBS评分显著降低(P<0.05);与正常组比较,模型组大鼠PI3K,Akt,m TOR mRNA的含量,以及PI3K,Akt,mTOR蛋白免疫阳性细胞数和表达量均升高(P<0.05);与模型组比较,给药组大鼠的上述指标均显著升高(P<0.05);与给药组比较,信号通路阻断组大鼠的上述指标均显著降低(P<0.05);上述指标在模型组与空白组大鼠之间的差异不具有统计学意义。结论:丹参注射液有助于大鼠SCI后脊髓神经功能的恢复,其机制可能与升高PI3K/Akt/mTOR信号转导通路的活性有关。
Objective: To investigate the mechanism of Salviae miltiorrhizae injection on spinal cord nerve function recovery after spinal cord injury (SCI). Methods: Rats (clean grade) were randomly divided into normal group, model group, administration group, blockade of signal pathway and blank group (n = 10). Except for the normal group, the other groups of rats were established gravity-hit SCI model. The rats in the model group were routinely housed after injury. The rats in the drug-treated group were intraperitoneally injected with Salviae Miltiorrhizae (1 m L · kg -1) 1 h after injury, and the Salvia Miltiorrhiza injection (1 m L · Kg -1, including rapamycin 3 mg · kg -1). The rats in the blank group were intraperitoneally injected with normal saline (1 m L · kg -1). The above groups were 1 / d injection. Animals were sacrificed on day 14 after sacrifice. Neurological functional recovery was evaluated by joint behavior score (CBS) before sacrifice and the spinal cord tissue was collected at 1 cm above and below the injured segment. The expression of PI3K, Akt and mTOR protein and mRNA in each experimental group were detected by immunohistochemical SP method, Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Results: Compared with the model group, the signal blockade group and the blank control group, the CBS score of the treated group was significantly decreased (P <0.05). Compared with the normal group, the expression of PI3K, Akt, m TOR mRNA (P <0.05). Compared with the model group, the above indexes in the treated group were significantly increased (P <0.05), and the levels of PI3K, Akt and mTOR protein were significantly increased Compared with the control group, the above indexes in the signal blockade group were significantly decreased (P <0.05). There was no significant difference in the above indexes between the model group and the blank group. Conclusion: Salvia miltiorrhiza injection can help the recovery of spinal nerve function after SCI in rats, which may be related to the activity of PI3K / Akt / mTOR signal transduction pathway.