Protein Interaction Between NL1 And Glutamate Receptors During LTP Damage Induced By Aluminum

Objective:To investigate the effect of subchronic exposure to aluminum on the changes of NL1,NX,NR1,NR2 A,NR2B and m Glu R protein,and to improve the biological mechanism of LTP induced by aluminum.Methods:(1)180 SD male rats were exposed to Al for 1,2,3 months,a total of 3 batches,and each batch of 60 rats were randomly divided into blank control group,saline control group and three Al(mal)3 exposure groups:high dose group(1.62mg/kg),middle dose group(0.81mg/kg)and low dose group(0.41mg/kg),and each dose group has 12 rats.Except the blank control group rats,others are all in sub chronic aluminum exposure model with intraperitoneal injection quaque die alterna;(2)The hippocampal CA1 area in hippocampus of rats was detected by electrophysiological method(LTP);(3)Graphite furnace atomic absorption spectrometry was used to determine the level of aluminum in hippocampus of rats;(4)Co immunoprecipitation and Western-blot assay was used to determine NX,NR1,NR2 A,NR2B,m Glu R1 protein expression in combination with NL1.Results:1 The level of aluminum in hippocampus of rats: Analysis of variance for aluminum exposure time and dose on brain aluminum levels in rats,and the results showed that compared to aluminum exposure for 1 months,brain aluminum levels increased obviously in the 2 months and 3 months exposed group(P<0.05),;Compared with the blank control group and physiological saline control group,low dose group,middle dose group and high dose group of brain aluminum levels were significantly increased(P<0.05).2.Effect of aluminum exposure on LTP in hippocampus of rats:(1)the results of LTP analysis in the aluminum exposure time and dose variance analysis results: The results showed that compared with the blank control group,LTP in the saline control group,low-dose group,middle dose group and high dose group were significantly lower(P<0.05).The LTP amplitude of high dose group was significantly lower than that of low dose group,middle dose group and saline control group(P<0.05).(2)multivariate repeated measures analysis of variance in each exposure time group LTP: The variance analysis of repeated measures of LTP in each dose group was performed in 1 month group,2 month group and 3 month group,the results showed that compared with the blank control group,the middle dose group and the high dose group of LTP were significantly lower(P<0.05).3.The expression of NL1 protein in hippocampus of rats: Analysis of variance and variance of NL1 protein expression in hippocampus of rats exposed to aluminum,The results showed that compared with the blank control group and saline control group,the expression of NL1 protein in low dose group,middle dose group and high dose group were significantly lower(P<0.05),However,there was no significant difference among the high dose group,the middle dose group and the low-dose group(P>0.05).4.The expression of NX protein in combination with NL1 antibody: Analysis of variance and variance of NX protein expression in combination with NL1 antibody by aluminum exposure time and dose,the results showed that the protein binding capacity of NX in the 2month and the 3 month group was significantly lower than that in 1 month group(P<0.05);.Compared with the blank control group and saline control group,the protein binding capacity of NX in low dose group,middle dose group and high dose group was significantly lower(P<0.05).The binding capacity of NX in high dose group was significantly lower than that in low dose group(P<0.05).5.The expression of glutamate receptors binding to NL1 antibody:(1)determination of NR1 protein binding on NL1:Analysis of variance and variance of NR1 protein expression in combination with NL1 antibody by aluminum exposure time and dose,the results showed that the protein binding capacity of NR1 in the 2 month and the 3 month group was significantly lower than that in the 1 month group(P<0.05);Compared with the blank control group and saline control group,the protein binding capacity of NR1 in low dose group,middle dose group and high dose group was significantly lower(P<0.05).The binding capacity of NR1 in high dose group was significantly lower than that in low dose group and medium dose group(P<0.05).(2)determination of NR2 A protein binding on NL1:Analysis of variance and variance of NR2 A protein expression in combination with NL1 antibody by aluminum exposure time and dose,the results showed that NR2 A protein binding rate was significantly decreased in the 2 month and the 3 month group compared with the 1 month group(P<0.05);Compared with the blank control group and saline control group,the protein binding capacity of NR2 A in low dose group,middle dose group and high dose group was significantly lower(P<0.05),The binding capacity of NR2 A in high dose group was significantly lower than that in low dose group and medium dose group(P<0.05).(3)determination of NR2 B protein binding on NL1:Analysis of variance and variance of NR2 B protein expression in combination with NL1 antibody by aluminum exposure time and dose,the results showed that compared with the 1 month group exposed to aluminum,the protein binding capacity of NR2 B was significantly decreased in 2 month and 3 months(P<0.05);Compared with blank control group and saline control group,low dose group,middle dose group and high dose group NR2 B protein binding amount was significantly reduced(P<0.05),the binding capacity of NR2 B in high dose group was significantly lower than that in low dose group and medium dose group(P<0.05).(4)determination of m Glu R1 protein binding on NL1:Analysis of variance and variance of m Glu R1 protein expression in combination with NL1 antibody by aluminum exposure time and dose,the results showed that compared with the 1 month group exposed to aluminum,the binding of m Glu R1 protein was significantly reduced in 3 months(P<0.05);Compared with blank control group and saline control group,low dose group,middle dose group and high dose group m Glu R1 protein binding amount was significantly reduced(P<0.05),the binding capacity of m Glu R1 in high dose group was significantly lower than that in low dose group and medium dose group(P<0.05).Conclusions:1.The decrease of LTP in hippocampal CA1 area of rats was induced by intraperitoneal injection of aluminum,the LTP amplitude decreased with increasing exposure time and dose.2.Reduction of NX binding to NL1 in hippocampus of rats exposed to aluminum,it is suggested that aluminum may affect the normal biological function of NL1 by disrupting the normal connections.3.Reduction of NMDAR and m Glu R binding to NL1 in hippocampus of rats exposed to aluminum,it is suggested that aluminum may disrupt protein-protein interactions.It is suggested that aluminum may disrupt protein-protein interactions.