通窍活血汤含药脑脊液抑制星形胶质细胞机械性损伤后炎症反应的机制研究  

Mechanisms of Tongqiao Huoxue Decoction-containing cerebrospinal fluid inhibiting inflammatory response of astrocytes after mechanical injury

作　　者：刘飞飞 申友奎[1] 陶开亮 张言涛 LIU Feifei;SHEN Youkui;TAO Kailiang;ZHANG Yantao(Department of Neurosurgery,Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou 310007,China;不详)

机构地区：[1]浙江中医药大学附属杭州市中医院神经外科,310007 [2]浙江中医药大学附属杭州市中医院普外科,310007

出　　处：《浙江医学》2024年第1期1-8,I0003,共9页Zhejiang Medical Journal

基　　金：浙江省中医药科技计划项目(2021ZA105、2023ZL545)。

摘　　要：目的探讨通窍活血汤含药脑脊液(TQHXD-CSF)对星形胶质细胞机械性损伤后炎症反应的影响以及其调控IL-33/肿瘤发生抑制蛋白(ST)2信号通路相关机制。方法将体外培养的星形胶质细胞按随机数字表法分为正常组(含20%正常脑脊液)、模型组(含20%正常脑脊液)、TQHXD-CSF组(含20%TQHXD-CSF)和跨膜型ST2(ST2L)中和抗体+TQHXD-CSF组(含20%TQHXD-CSF+5、10、20μg/mL ST2L中和抗体);除正常组外,其余各组均按星形胶质细胞机械性损伤模型构建方法处理细胞。在倒置显微镜下观察细胞形态,四甲基偶氮唑盐法检测细胞活性,ELISA法检测细胞上清液IL-33、ST2L、IL-1β及转化生长因子β1(TGF-β1)水平,Western blot、qRT-PCR法分别检测细胞IL-33、ST2L蛋白及mRNA表达水平。结果与正常组比较,模型组星形胶质细胞出现反应性增生改变,细胞活性及TGF-β1水平均明显降低(均P<0.05),IL-33、ST2L、IL-1β水平以及IL-33、ST2L蛋白及mRNA表达水平均明显升高(均P<0.05);与模型组比较,TQHXD-CSF组细胞形态明显改善,细胞活性及TGF-β1水平均明显升高(均P<0.05),IL-33、ST2L、IL-1β水平以及IL-33、ST2L蛋白及mRNA表达水平均明显降低(均P<0.05)。与TQHXD-CSF组比较,ST2L中和抗体(5、10、20μg/mL)+TQHXD-CSF组IL-1β水平均明显降低(均P<0.05),ST2L中和抗体(10、20μg/mL)+TQHXD-CSF组TGF-β1水平均明显升高(均P<0.05)。结论TQHXD-CSF能够抑制星形胶质细胞机械性损伤后的炎症反应,其作用机制可能与下调IL-33/ST2信号通路有关。Objective To investigate the effect of Tongqiao Huoxue Decoction-containing cerebrospinal fluid(TQHXDCSF)on the inflammatory response of astrocytes after mechanical injury and its regulation of IL-33/suppressor of tumorigenicity(ST)2 signaling pathway.Methods In vitro astrocytes were randomly divided into normal group(containing 20% normal cerebrospinal fluid),model group(containing 20% normal cerebrospinal fluid),TQHXD-CSF group(containing 20%TQHXD-CSF)and transmembrane ST2(ST2L)neutralizing antibody+TQHXD-CSF group(containing 20% TQHXD-CSF+5,10,20μg/mL ST2L neutralizing antibody)according to the random number table method.Except for the normal group,the cells in the other groups were treated according to the method of astrocyte mechanical injury modeling.Cell morphology was observed under an inverted microscope,cell activity was detected by methyl thiazolyl tetrazolium assay,the levels of IL-33,ST2L,IL-1β and transforming growth factor-β1(TGF-β1)in the cell supernatant were detected by ELISA,and the protein and mRNA expression levels of IL-33 and ST2L were detected by Western blotting and qRT-PCR,respectively.Results Compared with the normal group,astrocytes in the model group showed reactive hyperplastic changes,the cell activity and TGF-β1 levels were significantly decreased(all P<0.05),while the levels of IL-33,ST2L,IL-1β,and the protein and mRNA expression levels of IL-33 and ST2L were significantly increased(all P<0.05).Compared with the model group,the TQHXDCSF group showed significantly improved cell morphology,significantly increased cell viability and TGF-β1 levels(all P<0.05),and significantly decreased IL-33,ST2L,IL-1β levels,and IL-33,ST2L protein and mRNA expressions(all P<0.05).Compared with the TQHXD-CSF group,the IL-1β levels in ST2L neutralizing antibody(5,10,20μg/mL)+TQHXD-CSF group were significantly decreased(all P<0.05),while the TGF-β1 levels in ST2L neutralizing antibody(10,20μg/mL)+TQHXDCSF group were significantly increased(all P<0.05).Conclusion TQHXD-CSF can inhibit the in

关 键 词：通窍活血汤 星形胶质细胞 含药脑脊液 白细胞介素33 肿瘤发生抑制蛋白2 信号通路 炎症反应 

分 类 号：R285[医药卫生—中药学]