Effects of Wutou Decoction on regulation of lncRNA NEAT1 in regulating the IRE1α/XBP1 pathway to relieve endoplasmic reticulum stress in chondrocyte
Objective:To investigate the effect and mechanism of Wutou Decoction in alleviating endoplasmic reticulum stress in thapsigargin(TG)induced chondrocytes through the lncRNA NEAT 1-controlled IRE1α/XBP1 pathway.Methods:Fluorescent in situ hybridization(FISH)was used to detect the distribution and expression of lncRNA NEAT1 in 25 μmol/L TG-induced chondrosinocytes;lncRNA NEAT1 silencing chondrocytes were constructed,and Real-time PCR was used to detect IRE1α and XBP1 mRNA levels.Western Blot was used to detect its IRE1α and XBP1 protein expression.The first generation of chondrocytes was used for follow-up experiments,blank group(10％DMEM),TG group(25 μmol/L TG+10％DMEM),Wutou Decoction with medicinal serum group(25 μmol/L TG+different concentrations containing serum)were set up.Real-time PCR was used to detect the level of lncRNA NEAT1 mRNA in TG-induced chondrocytes;Western Blot was used to detect the expression of IRE1α,XBP1,EDEM1,and CHOP proteins in TG-induced chondrocytes.Immunofluorescence staining observed the fluorescence expression of IRE1α and Caspase-12 in each group of chondrocytes.Flow cytometry measures the apoptosis rate of chondrocytes in each group.Results:The FISH results showed that lncRNA NEAT1 was mainly distributed in the nucleus of chondrocytes.Compared with the TG+Lenti-control group,the mRNA levels and protein expression of IRE1α,XBP1 in the TG+sh-NEAT1 group were significantly reduced(P＜0.05,P＜0.01).Compared with the TG group,the levels of lncRNA NEAT1 and the expression of IRE1α,XBP1,EDEM1 and CHOP proteins in Wutou Decoction with medicinal serum group were significantly reduced(P＜0.01),the fluorescence expression of IRE1α and Caspase-12 was significantly reduced(P＜0.05),and the apoptosis rate of chondrocytes was significantly reduced(P＜0.05).Conclusion:Through lncRNA NEAT1 regulating IRE1α/XBP1 pathway,Wutou Decoction can relieve endoplasmic reticulum stress induced by TG and delay the degeneration of chondrocytes.
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