Glioma is one of the lethal central nervous system tumors.The infiltrative and invasive growth nature makes it difficult to identify the boundary between glioma and the normal tissues,resulting in inevitable recurrenc...Glioma is one of the lethal central nervous system tumors.The infiltrative and invasive growth nature makes it difficult to identify the boundary between glioma and the normal tissues,resulting in inevitable recurrence after surgery operation.Gliomas do not metastasize,so to prevent the residual tumor from proliferating or invading is a key challenge.Previous report indicated that hypotaurine could facilitate glioma invasion and suppress demethylases’activities.Using a hypotaurine synthesis deficient U251 cell line,we proved that the cells invasion ability was impaired.Gene expression profile analysis exhibited that knocking down one of the key enzymes of hypotaurine synthesis,2-aminoethanethiol dioxygenase(ADO),significantly affected the extracellular matrix-receptor process.Of that process,Wnt5a expression was severely upregulated by decreased intracellular ADO expression.Cells cultured at the presence of hypotaurine showed a decrease in intracellular Wnt5a protein and mRNA levels.This phenotype was due to hypermethylation of Wnt5a promoter,which was most likely the result of hypotaurine’s inhibiting demethylases activities.Collectively,this study demonstrated that hypotaurine synthesis deficient U251 cells were prone to epigenetic modification and Wnt5a seemed to be a tumor suppressor under that circumstance.This tumor suppression effect is warranted to be reevaluated in real tumor samples and the relevant evidence might contribute to develop new glioma interference strategies.展开更多
BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of tra...BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of traditional Chinese medicine,Ji-Chuan decoction(JCD)has been extensively and effectively used in STC treatment,yet its pharmacological mechanism remains unclear.AIM To explore the integrated regulatory pattern of JCD against STC through hyphenated techniques from metabolism,network pharmacology and molecular methods.METHODS STC model mice were generated by intragastric administration of compound diphenoxylate(10 mg/kg/d)for 14 d.The STC mice in the low dose of JCD(3.04 g/kg),middle dose of JCD(6.08 g/kg)and high dose of JCD(12.16 g/kg)groups were orally administered JCD solution once a day for 2 wk.The acetylcholine(ACH)level was examined by enzyme-linked immunosorbent assay.The pathological features of colon tissue were observed by hematoxylin and eosin staining.The differentially expressed metabolites and metabolic pathways were tested by nontargeted metabolomics.The main targets and core ingredients of JCD were identified by network pharmacology,and the expression of AKT was confirmed by immunohistochemistry.Finally,the pathways involved in JCD treatment were predicted using a combination of differentially expressed metabolites and targets,and intestinal glial cell apoptosis was demonstrated by immunofluorescence.RESULTS JCD significantly promoted intestinal motility,increased the levels of the excitatory neurotransmitter ACH and reduced intestinal inflammation in STC mice.Untargeted metabolomics results showed that JCD significantly restored metabolic dysfunction and significantly affected taurine and hypotaurine metabolism.Network pharmacology and molecular experiments showed that JCD regulates AKT protein expression,and the core component is quercetin.Combined analysis demonstrated that apoptosis may be an important mechanism by which JCD relieves constipation.Further experiments showed that JCD reduced enteric glial cell(EGC)apoptosis.CONCLUSION This work demonstrated that reducing EGC apoptosis may be the critical mechanism by which JCD treats STC.These findings call for further molecular research to facilitate the clinical application of JCD.展开更多
基金supported by New and Old Energy Conversion Project of Jining City(2017ZDGH031)Guiding funds for the development of local science and technology by the Central government(2017106014)Natural Science Foundation of Liaoning Province(L2015317).
文摘Glioma is one of the lethal central nervous system tumors.The infiltrative and invasive growth nature makes it difficult to identify the boundary between glioma and the normal tissues,resulting in inevitable recurrence after surgery operation.Gliomas do not metastasize,so to prevent the residual tumor from proliferating or invading is a key challenge.Previous report indicated that hypotaurine could facilitate glioma invasion and suppress demethylases’activities.Using a hypotaurine synthesis deficient U251 cell line,we proved that the cells invasion ability was impaired.Gene expression profile analysis exhibited that knocking down one of the key enzymes of hypotaurine synthesis,2-aminoethanethiol dioxygenase(ADO),significantly affected the extracellular matrix-receptor process.Of that process,Wnt5a expression was severely upregulated by decreased intracellular ADO expression.Cells cultured at the presence of hypotaurine showed a decrease in intracellular Wnt5a protein and mRNA levels.This phenotype was due to hypermethylation of Wnt5a promoter,which was most likely the result of hypotaurine’s inhibiting demethylases activities.Collectively,this study demonstrated that hypotaurine synthesis deficient U251 cells were prone to epigenetic modification and Wnt5a seemed to be a tumor suppressor under that circumstance.This tumor suppression effect is warranted to be reevaluated in real tumor samples and the relevant evidence might contribute to develop new glioma interference strategies.
基金Supported by the National Natural Science Foundation of China,No. 82074151the Experimental Formulary Sichuan Youth Science and Technology Innovation Research Team,No. 2020JDTD0022
文摘BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of traditional Chinese medicine,Ji-Chuan decoction(JCD)has been extensively and effectively used in STC treatment,yet its pharmacological mechanism remains unclear.AIM To explore the integrated regulatory pattern of JCD against STC through hyphenated techniques from metabolism,network pharmacology and molecular methods.METHODS STC model mice were generated by intragastric administration of compound diphenoxylate(10 mg/kg/d)for 14 d.The STC mice in the low dose of JCD(3.04 g/kg),middle dose of JCD(6.08 g/kg)and high dose of JCD(12.16 g/kg)groups were orally administered JCD solution once a day for 2 wk.The acetylcholine(ACH)level was examined by enzyme-linked immunosorbent assay.The pathological features of colon tissue were observed by hematoxylin and eosin staining.The differentially expressed metabolites and metabolic pathways were tested by nontargeted metabolomics.The main targets and core ingredients of JCD were identified by network pharmacology,and the expression of AKT was confirmed by immunohistochemistry.Finally,the pathways involved in JCD treatment were predicted using a combination of differentially expressed metabolites and targets,and intestinal glial cell apoptosis was demonstrated by immunofluorescence.RESULTS JCD significantly promoted intestinal motility,increased the levels of the excitatory neurotransmitter ACH and reduced intestinal inflammation in STC mice.Untargeted metabolomics results showed that JCD significantly restored metabolic dysfunction and significantly affected taurine and hypotaurine metabolism.Network pharmacology and molecular experiments showed that JCD regulates AKT protein expression,and the core component is quercetin.Combined analysis demonstrated that apoptosis may be an important mechanism by which JCD relieves constipation.Further experiments showed that JCD reduced enteric glial cell(EGC)apoptosis.CONCLUSION This work demonstrated that reducing EGC apoptosis may be the critical mechanism by which JCD treats STC.These findings call for further molecular research to facilitate the clinical application of JCD.
