芍药苷通过抑制氧化应激及凋亡保护高糖诱导大鼠皮质神经元损伤  

Paeoniflorin protects rats against high blood glucose-induced injury of cortical neurons by inhibiting their oxidative stress and apoptosis

作　　者：吴琼 崔利 张大伟 马斌斌 Wu Qiong;Cui Li;Zhang Dawei;Ma Binbin(Department of Neurology,Dalian Central Hospital,Dalian 116000,Liaoning Province,China)

机构地区：[1]大连医科大学附属大连市中心医院神经内科,116000 [2]大连医科大学附属第二医院神经外科

出　　处：《中华老年心脑血管病杂志》2022年第1期82-86,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81902541)。

摘　　要：目的探讨芍药苷对高糖诱导大鼠皮质神经元氧化应激及凋亡的保护作用。方法从新生SD乳鼠大脑皮质分离和培养神经元,4 d后分为对照组、高糖组(50 mmol/L高糖)、低、中和高浓度组(2.5、5.0、10.0μmol/L芍药苷预处理再加50 mmol/L高糖)。采用细胞增殖活力检测试剂观察细胞活力,用吸光度(A)值表示;检测超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;通过2',7'-二氢二氯荧光素二乙酸酯荧光探针检测活性氧水平;Hoechst 33342染色检测细胞凋亡;免疫荧光染色测定细胞线粒体膜电位(MMP);Western blot测定Bcl-2、Bax、裂解的半胱氨酸天冬氨酸蛋白酶3(cleaved-Caspase3)蛋白表达。结果与对照组比较,高糖组A值明显降低(0.452±0.505 vs 0.985±1.010,P<0.01)。与高糖组比较,低浓度组A值无明显变化(P>0.05),中和高浓度组A值增高(0.648±0.714和0.782±0.805 vs 0.452±0.505,P<0.05,P<0.01);SOD、CAT活性增加,活性氧水平降低,MMP表达增加;凋亡细胞减少;且中和高浓度组Bcl-2蛋白(0.506±0.601和0.742±0.803 vs 0.107±0.137)表达显著增加、Bax(0.748±0.659和0.411±0.355 vs 1.488±1.359)、cleaved-Caspase3(1.274±1.438和1.036±0.853 vs 2.137±2.601)蛋白表达降低(P<0.05,P<0.01)。结论芍药苷能够抑制高糖诱导的乳鼠皮质神经元细胞氧化应激反应及凋亡,其作用可能与调节凋亡相关蛋白有关。Objective To study the role of paeoniflorin(PF)in protecting rats against high glucose(HG)-induced oxidative stress and apoptosis of cortical neurons.Methods Cortical neurons were divided into control group,50 mmol/L HG group,2.5μmol/L PF pretreatment group,5μmol/L PF pretreatment group,and 10μmol/L PF pretreatment group 4 days after they were isolated from the cerebral cortex of newborn SD rats and cultured.The viability of cortical neurons was assayed with CCK8 and presented as absorbance(A).The activity of SOD and CAT was tested.The activated oxygen level was measured with a fluorescent probe.The mitochondrial membrane potential(MMP)and apoptosis of cortical neurons were detected with immunofluorescence stai-ning and Hoechst 33342 staining respectively.The expressions of Bcl-2,Bax,and cleaved-Caspase3 proteins were detected by Western blot.Results The A value was lower in 50 mmol/L HG group than in control group(0.452±0.505 vs 0.985±1.010,P<0.01).No significant change in viabil-ity of cortical neurons was detected between 50 mmol/L HG group and 2.5μmol/L PF pretreat-ment group(P>0.05).The A value was higher in 5μmol/L PF pretreatment group and 10μmol/L PF pretreatment group than in 50 mmol/L HG group(0.648±0.714 and 0.782±0.805 vs 0.452±0.505,P<0.05,P<0.01).The activity of SOD and CAT was higher,and serum level of activated oxygen was lower while the MMP expression level was higher and the number of apoptosed cortical neurons was smaller,the expression levels of Bcl-2(0.506±0.601 and 0.742±0.803 vs 0.107±0.137)were higher while those of Bax(0.748±0.659 and 0.411±0.355 vs 1.488±1.359)and cleaved-Caspase3(1.274±1.438 and 1.036±0.853 vs 2.137±2.601)were lower in 5μmol/L PF pretreatment group and 10μmol/L PF pretreatment group than in 50 mmol/L HG group(P<0.05,P<0.01).Conclusion PF inhibits the HG-induced oxidative stress and apoptosis of cortical neurons in newborn rats by regulatiing the apoptosis-related proteins.

关 键 词：芍药甙 氧化性应激 细胞增殖 神经元 超氧化物歧化酶 过氧化氢酶 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]