右美托咪定对高糖诱导大鼠乳鼠海马神经元损伤的保护作用

Protective effects of dexmedetomidine on the hippocampal neurons injuru induced by high glucose in neonatal rats

目的探讨右美托咪定对高糖诱导大鼠乳鼠海马神经元损伤的保护作用及机制。方法原代培养、鉴定SD大鼠乳鼠海马神经元,取其细胞并培养,将神经元细胞混悬液分为对照组、高糖组(给予100 mmol/L葡萄糖处理24 h)、高糖+右美托咪定组(400μmol/L右美托咪定与100 mmol/L葡萄糖共同处理24 h),CCK-8检测3组细胞活力,TUNEL实验检测3组细胞凋亡情况,免疫荧光检测3组突触素(SYP)蛋白表达,WB检测SYP蛋白、凋亡蛋白(cleaved cas-3、Bcl-2和Bax)以及PI3K/Akt和JAK2/STAT3信号通路蛋白表达。结果与对照组比较,高糖组神经元细胞的存活率显著降低(P<0.05);与高糖组比较,高糖+右美托咪定组神经元细胞的存活率显著升高(P<0.05)。与对照组相比,高糖组神经元细胞的凋亡率以及促凋亡蛋白cleaved cas-3、Bax显著升高,抑制凋亡蛋白Bcl-2显著降低(P<0.05);与高糖组比较,高糖+右美托咪定组神经元细胞的凋亡率以及cleaved cas-3、Bax蛋白显著降低,Bcl-2显著升高(P<0.05)。与对照组比较,高糖组神经元细胞SYP、p-Akt、p-STAT3蛋白显著降低(P<0.05);与高糖组比较,高糖+右美托咪定组神经元细胞SYP、p-Akt、p-STAT3蛋白显著升高(P<0.05)。结论右美托咪定可以逆转高糖诱导的大鼠乳鼠海马神经元凋亡,对神经元损伤发挥保护作用,机制可能与激活PI3K/Akt和JAK2/STAT3信号通路相关。

Objective To investigate the protective effects of dexmedetomidine on the hippocampal neurons injuru induced by high glucose in neonatal rats,and to explore its action mechanism.Methods The hippocampal neurons of SD suckling mice were cultured via primary culture,and neuron specific protein NeuN of hippocampal neurons was identified by immunofluorescence detection.Hippocampal neurons were divided into three groups:control group(without treatment),high glucose group(treated by 100mmol/L glucose for 24h)and high glucose+dexmedetomidine group(treated by 400mol/L dexmedetomidine and 100mmol/L glucose for 24h).The cell vitality and apoptosis rate were detected by CCK-8 and Tunel test,respectively,and the expression levels of synaptophysin(SYP)protein were detected by immunofluorescence detection.Moreover the expression levels of SYP protein,apoptosis protein:cleaved cas-3,Bcl-2,Bax,PI3K/Akt and JAK2/STAT3 signaling pathway associated protein were detected by Western Blot.Results Compared with tthat in control group,the survival rate of neurons in high glucose group was significantly decreased(P<0.05),however,compared with the high glucose group,the survival rate of neurons in high glucose+dexmedetomidine group was significantly increased(P<0.05).Compared with those in control group,the apoptosis rate and the levels of apoptosis promoting proteins cleaved cas-3 and Bax in high glucose group were significantly increased,while the expression levels of Bcl-2 were decreased significantly(P<0.05).Compared with those in high glucose group,the apoptosis rate,and the levels of cleaved CAS-3 and Bax protein of neurons in high glucose+dexmedetomidine group decreased significantly,while Bcl-2 levels were increased significantly(P<0.05).Compared with those in control group,the levels of SYP,p-Akt and p-STAT3 protein in high glucose group were decreased significantly(P<0.05).Compared with those in high glucose group,the levels of SYP,p-Akt and p-STAT3 protein in high glucose+dexmedetomidine group were significantly increased(P<0.05).Conclusion Dexmedetomidine can reverse the high glucose induced apoptosis of hippocampal neurons in neonatal rats,and its action mechanism may be associated with the activation of PI3K/Akt and JAK2/STAT3 signaling pathways.