大鼠交泰丸含药血清预处理对人脑胶质细胞氧化损伤的改善作用及其分子机制  

Ameliorative effect of pretreatment with medicated serum of Jiaotai pills on oxidative damage of human brain glial cells in rats and its molecular mechanism

作　　者：区浩松 杨阳 赵俊云 OU Haosong;YANG Yang;ZHAO Junyun(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,北京102488

出　　处：《山东医药》2022年第17期43-46,共4页Shandong Medical Journal

基　　金：北京中医药大学横向课题项目(2170071720009)。

摘　　要：目的观察大鼠交泰丸含药血清预处理对人脑胶质细胞(HEB)氧化损伤的改善作用并探讨其分子机制。方法SD大鼠随机分为两部分,分别通过肉桂及黄连水提物灌胃、同体积蒸馏水灌胃制备交泰丸含药血清及空白血清,将DMEM培养基与血清配制成10%完全培养基。将HEB分为对照组、模型组及交泰丸组,对照组及模型组加入空白血清培养基、交泰丸组加入含药血清培养基。培养24 h后,交泰丸组及模型组使用过氧化氢处理建立HEB氧化损伤模型,对照组使用等体积PBS处理。采用β-半乳糖苷酶染色法观察各组细胞氧化损伤比例,CCK-8法观察细胞活力,流式细胞术观察细胞凋亡率,活性氧荧光染色法观察细胞氧化应激水平,qRT-PCR法检测细胞衰老相关基因沉默信息调节因子1(SIRT1)、趋化因子联接因子1(CXCL1)、叉头框转录因子O亚族1(FOXO1)、P16、白细胞介素1α(IL-1α)、胰岛素样生长因子结合蛋白3(IGFBP3)、IL-6 mRNA表达。结果细胞氧化损伤比例模型组>交泰丸组>对照组,细胞活力对照组>交泰丸组>模型组,细胞凋亡率模型组>交泰丸组>对照组(P均<0.05)。对照组细胞核膜边界清晰,ROS荧光强度低;模型组细胞核膜发生部分裂解,ROS荧光强度增强;交泰丸组细胞膜及核膜结构较模型组更加完整,ROS荧光强度降低。与对照组比较,模型组SIRT1、FOXO1 mRNA表达下降,CXCL1 mRNA表达上升;与模型组比较,交泰丸组SIRT1、FOXO1 mRNA表达下降(P均<0.05),三组其他基因比较差异均无统计学意义。结论大鼠交泰丸含药血清预处理可抑制过氧化氢诱导的HEB氧化损伤,其机制可能与增强细胞活力、抗细胞凋亡、减少细胞氧化应激水平以及抗炎症有关。Objective To observe the ameliorative effect of pretreatment with medicated serum of Jiaotai pills on oxidative damage of human brain glial cells(HEB)in rats and to explore its molecular mechanism.Methods SD rats were randomly divided into two parts,and the medicated serum was prepared by gavage of the aqueous extract of cinnamon and coptis,and the blank serum was prepared by gavage of the same volume of distilled water.The DMEM medium and the serum were prepared into 10%complete medium.The HEB cells were divided into the control group,model group and Jiaotai pills group.The blank serum medium was added to the control group and model group,and the medicated serum medium was added to the Jiaotai pills group.After 24 h of cultivation,the HEB oxidative damage model was established by hydrogen peroxide treatment in the Jiaotai pill group and model group,and the control group was treated with the same volume of PBS.The proportion of cell oxidative damage in each group was observed by β-galactosidase staining,cell viability was detected by CCK-8,apoptosis rate was observed by flow cytometry,oxidative stress level was observed by reactive oxygen species(ROS)fluorescence staining,and the mRNA expression of cellular senescence-related genes,including sirtuin 1(SIRT1),C-X-C motif chemokine ligand 1(CXCL1),forkhead box O1(FOXO1),cyclin dependent kinase inhibitor 2A(P16),interleukin 1α(IL-1α),insulin like growth factor binding protein 3(IGFBP3),and interleukin 6(IL-6)was detected by qRT-PCR.Results The proportion of cell oxidative damage was the highest in the model group and the lowest in the control group,the cell viability was the highest in the control group and the lowest in the model group,and apoptosis rate was the highest in the model group and the lowest in the control group(all P<0.05).In the control group,the nuclear membrane boundary was clear and the ROS fluorescence intensity was low;in the model group,the nuclear membrane was partially cleaved and the ROS fluorescence intensity was enhanced;in the Jiaotai p

关 键 词：交泰丸 含药血清 人脑胶质细胞 氧化损伤 分子机制 

分 类 号：R285.5[医药卫生—中药学]