| ObjectiveA type of acid,taurine ursodeoxycholic acid(TUDCA),was found in the traditional Chinese medicine "bear bile".It is a hydrophilic bile acid that is naturally produced in the liver by conjugating taurine with ursodeoxycholic acid(UDCA).TUDCA can penetrate the blood-brain barrier and has low toxicity.It has neuroprotective effects in models of neurodegenerative diseases(ND),This study observed the effect of taurine ursodeoxycholic acid on spinal cord neuron apoptosis under conditions of glucose deficiency and hypoxia,as well as its effect on motor function recovery in mice after acute spinal cord injury.Methods1.In vitro experiment:Primary spinal cord neurons were isolated and extracted from C57 BL/6 mouse embryos at 13.5 days of gestation.After 72 hours of culture,they were divided into three groups for treatment:the normal group was added to Neurobasal complete culture medium and cultured in a normal incubator(volume fraction 5%CO2+95%N2)for 24 hours;The oxygen glucose deprivation group was added to sugar free Neurobasal medium and incubated in a three gas incubator(volume fraction 94%N2+5%CO2+1%O2)for 12 hours.The group was replaced with Neurobasal complete medium and then incubated in a CO2 incubator for 12 hours;The treatment process of the experimental group was roughly the same as that of the oxygen glucose deprivation group,where TUDCA was added while sugar free Neurobasal medium was added.TUNEL staining was used to detect cell apoptosis,CCK-8 method was used to detect cell activity,and immunofluorescence staining was used to detect cells β Ⅲ Microtubulin expression.2.Animal experiment:Sixty C57 BL/6 mice were randomly divided into a sham operation group,a spinal cord injury group,and an experimental group using a random number table method,with 20 mice in each group.The spinal cord injury group and the experimental group established a T9-T10 spinal cord injury model using Allen’s blow method.Starting from the first day after modeling,the experimental group was given TUDCA solution by gavage.The sham operation group and the spinal cord injury group were given physiological saline by gavage once a day.After continuous administration for 14 days,the repair of spinal cord tissue was evaluated using behavioral and histological methods.ResultsIn vitro experiments:TUNEL staining,CCK-8 staining,and immunofluorescence staining showed that the number of apoptotic cells in the oxygen glucose deprivation group was higher than that in the normal group(P<0.01),and cell activity was correlated with β The expression of microtubule protein III was lower than that of the normal group(P<0.01);The number of apoptotic cells in the experimental group was lower than that in the oxygen glucose deprivation group(P<0.01),and cell activity was correlated with β The expression of microtubule protein III was higher than that of the oxygen glucose deprivation group(P<0.01).Animal experiments:The BBB score and hindlimb footprint experiments in the open field experiment showed that the walking and motor function recovery of the experimental group mice was better than that of the spinal cord injury group.Hematoxylin eosin staining showed that in the spinal cord injury group of mice,obvious deformities and cavities were observed at the spinal cord injury site,and the number of nerve cells was significantly reduced;Compared with the spinal cord injury group,the experimental group showed a significant reduction in the lesion area of spinal cord injury,a smaller degree of spinal cord malformation,fewer cavities,and an increase in the number of nerve cells.Immunofluorescence staining showed that the number of labeled neurons in the spinal cord injury group was lower than that in the sham surgery group(P<0.01),while the number of labeled neurons in the experimental group was higher than that in the spinal cord injury group(P<0.01).ConclusionTUDCA can reduce spinal cord neuron apoptosis and axonal loss caused by glucose deficiency and hypoxia,and promote the recovery of motor function in spinal cord injury mice. |
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