Mechanisms of Tongqiao Huoxue Decoction-containing cerebrospinal fluid inhibiting inflammatory response of astrocytes after mechanical injury
Objective To investigate the effect of Tongqiao Huoxue Decoction-containing cerebrospinal fluid(TQHXD-CSF)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+5,10,20 μg/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-1 β 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-1 β,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 TQHXD-CSF 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-1 β levels,and IL-33,ST2L protein and mRNA expressions(all P＜0.05).Compared with the TQHXD-CSF group,the IL-1 β levels in ST2L neutralizing antibody(5,10,20 μg/mL)+TQHXD-CSF group were significantly decreased(all P＜0.05),while the TGF-β1 levels in ST2L neutralizing antibody(10,20 μg/mL)+TQHXD-CSF group were significantly increased(all P＜0.05).Conclusion TQHXD-CSF can inhibit the inflammatory response of astrocytes after mechanical injury,and its mechanism may be related to down-regulating IL-33/ST2 signaling pathway.
Tongqiao Huoxue DecoctionAstrocyteMedicated cerebrospinal fluidInterleukin-33Suppressor of tumourigenicity 2Signaling pathwayInflammation
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维普
