TRPC1在缺氧缺血性脑损伤新生大鼠脑皮层内的表达及头孢曲松钠的干预作用

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目的研究TRPC1在缺氧缺血性脑损伤(HIBD)新生大鼠脑皮层内的表达,初步探讨头孢曲松钠(CTX)对其影响及可能机制。方法将7日龄新生SD大鼠随机分为假手术组(S组)、生理盐水组(C组)及头孢曲松钠治疗组(CTX组)。C组、CTX组按照Rice法制备HIBD模型,每组均依术后观察时间点不同分为6h、12h、24h、3d、5d、7d共6个亚组,每个亚组5只,在相应时间点断头取脑采用Western印迹和免疫组化法检测各组病变侧脑组织TRPC1蛋白的表达变化。结果 Western印迹结果:C组TRPC1蛋白的表达水平在各时间点均较S组明显升高(P<0.05),TRPC1在HIBD后1 d达到高峰,随后开始下降。CTX组在每个观察时间点TRPC1的水平均低于C组(P<0.05),但高于S组(P<0.05)。免疫组化染色结果:S组在各时间点脑皮质中可见少量棕黄色颗粒阳性细胞,C组在HIBD后阳性细胞数明显高于S组,且在第1天达高峰,随后逐渐下降(P<0.05);CTX组表达趋势与C组类似(P<0.05),各时间点的阳性细胞数较S组高但均低于C组(P<0.05)。结论新生大鼠HIBD后病变侧大脑皮层TRPC1的表达水平明显升高,并具有时间差异性,表明其参与了HIBD的病理过程,可能在HIBD诱导的细胞凋亡中发挥重要作用。头孢曲松钠能够下调TRPC1的表达,从而起到对HIBD新生大鼠的脑保护作用。 Objective To investigate the expression of TRPC1 in cortex of neonatal rats with hypoxic-ischemic brain damage (HIBD) and to investigate the effect of CTX on it and its possible mechanism. Methods 7-day-old SD rats were randomly divided into sham operation group (S group), saline group (C group) and ceftriaxone sodium treatment group (CTX group). Group C and CTX were prepared according to the method of Rice HIBD model. Each group was divided into 6 subgroups at 6h, 12h, 24h, 3d, 5d and 7d according to the observation time points after operation. There were 6 subgroups in each subgroup, The brain was decapitated at the time point, and the expression of TRPC1 protein in the lesion side of each group was detected by Western blot and immunohistochemistry. Results Western blotting results showed that TRPC1 protein expression in group C was significantly higher than that in group S at each time point (P <0.05). TRPC1 peaked on the first day after HIBD and then began to decline. The level of TRPC1 in CTX group at each observation time point was lower than that in C group (P <0.05), but higher than that in S group (P <0.05). Immunohistochemical results: S group showed a small amount of brown granules positive cells in cerebral cortex at each time point. The number of positive cells in group C after HIBD was significantly higher than that in group S, and reached the peak on the first day, then decreased gradually (P <0.05). The expression of CTX group was similar to that of C group (P <0.05). The number of positive cells at each time point was higher than that of S group but lower than that of C group (P <0.05). Conclusion The expression of TRPC1 in the ischemic cortex of neonatal rats with HIBD is significantly increased with time difference, indicating that TRPC1 is involved in the pathological process of HIBD and may play an important role in HIBD-induced apoptosis. Ceftriaxone sodium can down-regulate the expression of TRPC1, thus playing a protective role in HIBD neonatal rats.
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