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目的:通过检测丹参注射液对大鼠脊髓损伤(SCI)后少突胶质细胞(Olig-2)和Ⅲ型补体受体(OX-42)系转录因子-2表达的影响,探讨其促进脊髓神经功能恢复的机制。方法:SPF大鼠50只随机分为正常组、模型组、丹参治疗组、丹参加雷帕霉素组和甲泼尼龙琥珀酸钠组(n=10)。采用Allen’s法建立SCI模型(正常组除外)后,模型组:腹腔注射生理盐水(1 m L·kg~(-1)),1次/d;丹参治疗组:腹腔注射丹参注射液(1 m L·kg~(-1)),1次/d;丹参加雷帕霉素组:腹腔注射同等剂量丹参注射液(含雷帕霉素3 mg·kg~(-1)),1次/d;甲泼尼龙琥珀酸钠组:尾静脉推注甲泼尼龙琥珀酸钠(30 mg·kg~(-1)),1次/d。伤后1,3,7,14 d时采用联合行为评分法(CBS)评价大鼠脊髓神经功能恢复情况。伤后14 d处死动物,采用免疫荧光和蛋白免疫印迹法(Western blot)检测各组大鼠脊髓内Olig-2和OX-42的表达。结果:至伤后14 d时,与模型组、丹参加雷帕霉素组比较,丹参治疗组和甲泼尼龙琥珀酸钠组大鼠CBS评分显著降低(P<0.05);与正常组比较,模型组大鼠Olig-2表达(免疫阳性细胞数及蛋白相对表达量)降低,而OX-42表达升高(P<0.05);与模型组比较,丹参治疗组和甲泼尼龙琥珀酸钠组Olig-2表达升高,而OX-42表达降低(P<0.05);与丹参治疗组和甲泼尼龙琥珀酸钠组比较,丹参加雷帕霉素组大鼠Olig-2表达降低,而OX-42表达升高(P<0.05);上述指标在丹参治疗组和甲泼尼龙琥珀酸钠组之间的差异不具有统计学意义。结论:丹参注射液可通过升高磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/m TOR信号转导通路的活性以调节Olig-2和OX-42的表达,从而表明其参与大鼠脊髓神经功能恢复的机制可能与通过参与少突胶质细胞和小胶质细胞增殖有关。
Objective: To investigate the effect of Danshen injection on the expression of transcription factor-2 in oligodendrocyte (Olig-2) and type Ⅲ complement receptor (OX-42) after spinal cord injury (SCI) Mechanism of neural function recovery. Methods: Fifty SPF rats were randomly divided into normal group, model group, Salvia miltiorrhiza group, Danshen plus rapamycin group and methylprednisolone sodium succinate group (n = 10). After establishing SCI model (except normal group) by Allen’s method, the model group was injected intraperitoneally with normal saline (1 m L · kg -1) L · kg ~ (-1)) once a day; Danshen plus rapamycin group: intraperitoneal injection of the same dose of Salvia injection (including rapamycin 3 mg · kg -1) d; methylprednisolone sodium succinate group: methylprednisolone sodium succinate (30 mg · kg -1) was injected through tail vein once a day. At 1, 3, 7 and 14 days after injury, the joint behavior score (CBS) was used to evaluate the recovery of spinal cord function in rats. The animals were sacrificed 14 days after injury, and the expression of Olig-2 and OX-42 in the spinal cord of each group was detected by immunofluorescence and Western blot. Results: Compared with the model group and Danshen plus rapamycin group, the CBS score of Salvia miltiorrhiza group and methylprednisolone sodium succinate group was significantly decreased (P <0.05) at the 14th day after injury. Compared with the normal group, Olig-2 expression (the number of immunopositive cells and protein relative expression) in the model group decreased, while the expression of OX-42 increased (P <0.05). Compared with the model group, the Salvia miltiorrhiza treatment group and methylprednisolone sodium succinate group Olig-2 expression increased, while OX-42 expression decreased (P <0.05); compared with the Salvia miltiorrhiza treatment group and the methylprednisolone sodium succinate group, Olig-2 expression in the Salvia plus rapamycin group decreased, while OX -42 (P <0.05). There was no significant difference in the above parameters between Salvia miltiorrhiza group and methylprednisolone sodium succinate group. Conclusion: Salvia miltiorrhiza injection can regulate the expression of Olig-2 and OX-42 by increasing the activity of PI3K / Akt / m TOR signal transduction pathway, The mechanism involved in the recovery of spinal cord nerve function may be related to the proliferation of microglia and microglia through their involvement.