参附注射液对急性创伤性脑损伤大鼠脑保护作用的研究

Protective effect of Shengfu injection on rats with acute traumatic brain injury

目的探讨参附注射液（SF）对大鼠急性创伤性脑损伤（ATBI）时谷氨酸转运体（GLT-1）和GLAST信号通路转运活性和表达水平，以及早期血清蛋白S100β、脑组织总钙含量和含水量的影响。方法清洁级健康雄性成年SD大鼠60只，体质量300～320g。采用随机数字表法分为三组（每组n=20）：假手术组（Sham组）、急性创伤性脑损伤组（ATBI组）和参附注射液处理组（SF组）。采用Feeney法重物自由落体脑损伤装置，制备大鼠脑损伤模型。sF处理组在制作脑损伤模型30min前经腹腔注射参附注射液20mL／kg，Sham组和ATBI组给予等容量生理盐水腹腔注射。于造模后15min（T1）和60min（T2）时检测血清s100β蛋白含量；大鼠造模4h后迅速断头处死，测定大鼠脑组织总钙含量及含水量；RT—PCR检测脑组织GLT-1和GLASTmRNA表达水平；采用蛋白质免疫印迹法（Western blot）检测GLT-1和GLAST蛋白的表达。结果与T1时比较，T2时ATBI与SF组血清S100β蛋白含量明显减少（P〈0．05）；与Sham组比较，ATBI组、SF组血清S100β蛋白含量、脑组织总钙含量及含水量明显增加，大鼠脑组织GLT-1、GLAST mRNA以及GLT-1和GLAST蛋白含量的表达明显减少（P〈0．05）。与ATBI组比较，SF组血清S100β蛋白含量、脑组织总钙含量及含水量明显减少，脑组织GLT-1、GLASTmRNA，以及GLT-1和GLAST蛋白含量的表达明显增加（P〈0．05）。结论参附注射液可减轻大鼠ATBI，其机制可能与其上调GLF-1活性和GLAST信号通路有关。

Objective To investigate the effects of Shenfu injection on expression level of glutamate transporter （GLT- 1 ） and GLAST signaling pathway, the early serum S100β protein, total calcium and water content in brain tissue of rats with acute traumatic brain injury. Methods Sixty male SD mice, weighing 300 - 320 g, were equally and randomly divided into 3 groups（ n = 20 each） using a random number table： sham group （group Sham）, acute traumatic brain injury group （group ATBI）, andshengfu injection group （group SF）. Models of ATBI were made according to Feeney＇s methods. Shengfu injection 20 mL/kg preconditioning was injected intraperitoneally 30 rain before making the model in group SF, while the equal volume of normal saline was given in Sham and ATBI groups. The serum S100B protein was measured at 15 min （T1） and 60 min （T2） after modeling. The rats were sacrificed 4 11 after modeling and the total calcium and water content were measured. GLT - 1 and GLAST mRNA were detected by RT - PCR, The expression of GLT - 1 and GLAST protein was detected by Western blot. Results Compared with T1, the content of SlOOp protein of T2 in ATBI and SF groups was significantly decreased （ P 〈 0. 05 ）. Compared with Sham group, the content of S100β protein, the total calcium and water content of brain tissue in ATBI and SF groups were significantly increased, and the expression of GLT - 1, GLAST mRNA and GLT - 1 and GLAST protein in brain tissue were significantly decreased （ P 〈 0. 05 ）. Compared with ATBI group, the contents of serum SlOOp protein, total calcium and water content in brain tissue were significantly decreased, and the expression of GLT - 1, GLAST mRNA and GLT - 1 and GLAST protein in brain tissue were significantly increased（P 〈0.05）. Conclusion Shenfu injection can relieve acute traumatic brain injury in rats, and its mechanism may be related with its up - regulation of glutamate transporter activity and GLAST signaling pathway.