| Chloroquine phosphate is a 4-aminoquinoline compound and is considered to be the first choice for the treatment of malaria.Chloroquine phosphate also has a spectral antiviral effect and has a therapeutic effect on some diseases caused by viruses.Although chloroquine phosphate has become a good antimalarial and antiviral drug,there are serious safety risks while using chloroquine phosphate to treat diseases.Therefore,it is of great significance to study the toxicity of chloroquine phosphate and provide scientific basis for the safe use of chloroquine phosphate in clinic.This experiment studied the toxicity mechanisms of several key tissues induced by chloroquine phosphate.Firstly,the toxic model of chloroquine phosphate in mice was established,multiple tissues(liver,heart,lung and brain)were detected by GC-MS,in order to identify the significantly altered metabolites and metabolic pathways between control and chloroquine phosphate treated groups,the GC-MS results were analyzed by orthogonal partial least squares and t-test using multivariate statistical analysis.Secondly,Western Blot was used to study the expression of LC3 B and NLRP3 in the chloroquine phosphate-induced hepatotoxicity model and the expression of NF-κB p65 in heart tissue.Finally,combined with Quantitative real-time PCR(qRT-PCR)to detect the expression of OPLAH gene and GGCT gene in the mouse lung tissue toxicity model induced by chloroquine phosphate.The results showed that this experiment identified7 metabolic markers with significant differences in liver tissues and found 8 related metabolic pathways,mainly related to the sugar metabolism pathway and fatty acid metabolism pathway,among them,galactose metabolism pathway was identified with significant differences,which may be the specific pathway of chloroquine phosphate mediated liver injury;In heart tissue,12 metabolic markers with significant differences were identified,and13 metabolic pathways were involved,mainly about amino acid metabolic pathway and inositol metabolic pathway;In lung tissue,7 metabolic markers with significant differences and 9 metabolic pathways related to them were identified,among which glutathione metabolic pathway has significant metabolic differences,which may be the specific pathway of chloroquine phosphate mediated lung injury;In the brain tissue,8 metabolic markers were identified with significant differences,and 13 metabolic pathways associated with them were identified.Among them,the aminoacyl-tRNA synthesis pathway and the metabolic pathways of glycine,serine and threonine had significant metabolic differences.Toxic metabolic markers and related metabolic pathways jointly induce the damage of chloroquine phosphate to various tissues.The toxicity mechanism of chloroquine phosphate is closely related to the changes of metabolic markers and the disorder of related metabolic pathways.The results of Western Blot showed that compared with the control group,the expression of NLRP3 increased and the expression of LC3 B decreased in the low-dose and high-dose groups.The changes of the expression of NLRP3 and LC3 B caused damage to the liver tissue.Compared with the control group,the protein expression of NF-κB p65 in the heart of mice in low-dose and high-dose groups was significantly increased,which may be an important reason for chloroquine phosphate induced cardiac injury.The results of qRT-PCR showed that the GGCT gene did not change significantly in the lung tissue of mice,and the expression of OPLAH gene in the low-dose and high-dose groups was significantly decreased compared with the control group,leading to the disorder of glutathione metabolism pathway and induced lung injury.This experiment elaborated the metabolic changes caused by chloroquine phosphate toxicity in several major tissues,and the mechanism of chloroquine phosphate induced toxicity was studied from protein and gene levels by combining Western Blot and qRT-PCR methods,which provided a new basis for elucidating the toxicity of chloroquine phosphate. |
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