摘要
目的:探讨柴胡疏肝散(CSS)通过调节自噬-溶酶体途径抑制β-淀粉样蛋白(Aβ)诱导神经毒性的作用机制。方法:选取大鼠PC12嗜铬细胞瘤细胞分化的类神经元细胞为研究对象,以Aβ诱导神经毒性为模型,采用噻唑蓝(MTT)法和乳酸脱氢酶(LDH)释放实验检测神经细胞毒性,采用单丹磺酰尸胺(MDC)染色和LysoTracker荧光探针染色观察细胞自噬,采用Western blot测定自噬-溶酶体途径相关蛋白的表达情况。结果:Aβ在0.125~2.0μmol·L^(-1)范围内呈浓度和时间依赖性抑制PC12细胞活力,而CSS在剂量250μg·mL^(-1)、500μg·mL^(-1)均明显抑制Aβ诱导的细胞活力下降及乳酸脱氢酶释放(P<0.05);荧光染色结果显示CSS激活Aβ诱导的细胞自噬和溶酶体活性;CSS可抑制细胞mTOR磷酸化和增加LC3-Ⅱ/LC3-Ⅰ水平,促进自噬体形成。结论:CSS抑制Aβ诱导的神经毒性,可能与其调节自噬-溶酶体途径有关。
Objective To explore the mechanism of Chaihu Shugan San(CSS)inhibitsβ-amyloid(Aβ)-induced neurotoxicity via regulating autophagy-lysosomal pathway.Methods The neuron-like cells differentiated from rat PC12 pheochromocytoma cells were selected as the research object,and the neurotoxicity induced by Aβwas used as the model.The cytotoxicity of nerve cells was detected by methyl thiazol tetrazolium(MTT)assay and lactate dehydrogenase(LDH)release assay.Monodansylcadaverine(MDC)staining and LysoTracker fluorescent probe staining were used to observe autophagy,and Western blot was used to detect the expression of autophagy-lysosome pathway-related proteins.Results Aβinhibited the viability of PC12 cells in a concentration and time-dependent manner within the range of 0.125-2.0μmol·L^(-1),while CSS at 250μg·mL^(-1)and 500μg·mL^(-1)significantly inhibited Aβ-induced decrease in cell viability and lactate dehydrogenase release(P<0.05).Fluorescence staining showed that CSS activated Aβ-induced autophagy and lysosomal activity.CSS can inhibit the phosphorylation of mTOR and increase the level of LC3-Ⅱ/LC3-Ⅰ,and promote the formation of autophagosome.Conclusion CSS inhibited Aβ-induced neurotoxicity probably by regulating autophagy-lysosomal pathway.
作者
曾嫱
李新萍
李利民
吴正治
ZENG Qiang;LI Xin-ping;LI Li-min;WU Zheng-zhi(The First Affiliated Hospital of Shenzhen University,Shenzhen Second People's Hospital,Guangdong Shenzhen 518035;Shenzhen Institute of Geriatrics,Guangdong Shenzhen 518035;The Eighth Affiliated Hospital of Sun Yat-sen University,Guangdong Shenzhen 518033)
出处
《深圳中西医结合杂志》
2022年第18期1-5,139,共6页
Shenzhen Journal of Integrated Traditional Chinese and Western Medicine
基金
广东省基础与应用基础研究基金项目(2020A1515010758)
深圳市基础研究专项(自然科学基金)基础研究项目(JCYJ20190812171611467)
深圳市福田区卫生健康系统科研项目(FTWS2022034)
作者简介
曾嫱,女,助理研究员,主要研究方向是中医药防治老年疾病;通信作者:吴正治,(E-mail:szwzz001@email.szu.edu.cn)
