Effect Of LncRNA-KCNQ1OT1 On The Prevention Of Neuronal Apoptosis After Acute Spinal Cord Injury By Zhenbao Pill

Objective Purpose: Acute spinal cord injury(ASCI)often leads to persistent organ dysfunction and permanent neurological deficits.Recent studies have found that Zhenbao pills can improve the ASCI condition of rats by inhibiting neuronal cell apoptosis.We used RNA chips to screen the difference in long non-coding RNA expression between Zhenbao pills treatment group and non-treatment group.The results showed that hypoxia treatment of mouse neuronal cells HT22(neural cell injury model)down-regulated the expression of KCNQ1OT1,Zhenbao pills can reverse the decline of KCNQ1OT1 in a dose-dependent manner,but the role of KCNQ1OT1 in neurons has not been reported.This study is to explore the role of KCNQ1OT1 in the treatment of acute spinal cord injury with Zhenbao Pills.Materials and methods An in vitro injury model of nerve cells caused by hypoxia treatment.Treat the injured cells with Zhenbao Pills and observe the changes in the expression level of KCNQ1OT1 after treatment with different doses of Zhenbao Pills.Treat the cells overexpressing KCNQ1OT1 and knockout KCNQ1OT1 with Zhenbao pills,and observe the changes of cell apoptosis.By preparing the acute spinal cord injury model in mice,the mice were divided into Zhenbao Pill group and control group,with 5 mice in each group.After the model was successfully established,Zhenbao pills were given by intragastric administration continuously to observe the expression of KCNQ1OT1 and the apoptosis of nerve cells in each group of mice.SPSS 20.0 statistical software was used for analysis.The count data were expressed as mean±standard deviation.Student’s t test was used to calculate the statistical difference between the two groups,and the measurement data was tested by chi-square test.One-way ANOVA was used to analyze the comparison between multiple groups.P<0.05 indicates that the difference is statistically significant.Result:1.In the mouse acute spinal cord injury model,it was found that the apoptosis of nerve cells in the Zhenbao Pill group was significantly reduced(P<0.01);2.Through gene chip screening and fluorescence quantitative PCR to detect the difference of Lnc RNA between Zhenbaowan group and Control group,it was found that the expression level of KCNQ1OT1 in Zhenbaowan group was significantly higher than that in the control group;3.In the in vitro injury model of nerve cells,it was found that hypoxia significantly down-regulated the expression of KCNQ1OT1(P<0.01);the pretreatment of Zhenbao Pills reversed its trend in a dose-dependent manner;4.The overexpression of KCNQ1OT1 significantly reduced the apoptosis of HT22 cells caused by hypoxia(P<0.01);5.Zhenbao Pills reduced the apoptosis of HT22 cells due to hypoxia(P<0.01);interference with KCNQ1OT1 partially cancelled the protective effect of Zhenbao Pills(P<0.01).Conclusion 1.KCNQ1OT1 is mainly expressed in the nucleus of neuronal cells.2.The expression of KCNQ1OT1 in the spinal cord injury area increased in the Zhenbao Pill treatment group.3.Zhenbao Pill reverses the decrease of KCNQ1OT1 expression in neuronal cells caused by hypoxia.4.Overexpression of KCNQ1OT1 reverses neuronal cell apoptosis caused by hypoxia.5.Interfere with KCNQ1OT1 to cancel the effect of Zhenbao Pill to protect neuronal cells from damage.