Electric For Samp8 Mice Plasma, Liver And Kidney ~ 1h Nmr Spectra Of The Impact

Object:To study the spectral alterations of the blood plasma,hepatic and renal tissue's metabolites in SAMP8 mice by using ¹H-NMR technique and the influence of the electro-acupuncture on them.At the same time,with an attempt to define the specific changes of the metabolites due to the effect of the electro-acupuncture.Methods:Sixteen SAMP8 mice were randomly divided into two groups(n=8):Model group and Electro-acupuncture group;SAMR1 mice used as Normal group(n=8).Acupoints "Baihui" and "Yongquan" were selected and treated with the stimulating power which kept the back legs of mice slightly shaken for 20min(thin-dense alternative wave, frequency 2-100HZ,electric volt 2-4V).once a day,7 days a cycle,three cycles(21 days) in total,1 day break between the cycles.¹H-NMR technique was used to obtain the small molecules' metabolic spectra of the blood plasma,extract of liver and kidney tissue,the data were normalized,integrated and treated by SIMCA-P10.0 software pack to analize the major component(PCA) and the partial leastquares discriminate (PLS-DA) in combination with OSC method,in this way,the corresponding score scattering plot and loading plot were obtained.Results:1.the result of ¹H NMR spectra from plasmaThe result of the CPMG experiment showed the obvious seperation among the three groups.The major chemical shifts which caused this seperation are at:δ1.34-δ1.38(lactate),δ1.22(3-hydroxybutyrate),δ1.3(fucose),δ1.42(alanine).The spectra of the Model group showed a decreased level of fucose(δ1.3)and 3-hydroxybutyrate(δ1.22) when compared with those of the Normal group;The spectra of the electro-acupuncture group in comparison with those of the Model group showed an increased level of lactate(δ1.34-δ1.38) and alanine(δ1.42).The result of LED experiment showed the obvious seperation among the three groups.The major chemical shifts which caused this seperation are at:δ0.87-δ0.91( CH₃-),δ1.31-δ1.39((CH₂)n),δ3.27(Phosphatidylcholine).The samples of the Model group compared with those of the Normal group contain less CH₃-(δ0.87), while the samples of the Electro-acupuncture group contain more CH₃-(δ0.91)and Phosphatidylcholine(δ3.27) than those of the Model group.Among these molecules, CH₃-(δ0.87-δ0.91) are from big molecules' protein mainly in the forms of HDL, LDL and VLDL.2.the result of ¹H NMR spectra from liverThe result of the water soluble extract showed the obvious seperation among the three groups.The major chemical shifts which caused this seperation are at:δ1.31(lipidtriglyceride),δ3.19-δ3.23(choline),δ3.27(taurine),δ3.43-δ3.47(taurine),δ3.79-δ3.87(glucose).In comparison with the Normal group,the Model group has higher level of glucose(δ3.79-δ3.87) and lower level of taurine(δ3.27,δ3.43-δ3.47); While the Electro-acupuncture group compared with the Model group has higher levels of choline(δ3.19-δ3.23) and lipid triglyceride(δ1.31),but lower level ofglucose(δ3.79-δ3.87).The result of the fat soluble extract showed the obvious seperation among the three groups.The major chemical shifts which caused this seperation are at:δ1.26-δ1.34((CH₂)_n-),δ2.02(glycoprotein),δ3.22(choline).The extract of the Model group compared with that of the Normal group has higher level of choline(δ3.22),but lower levels of(CH₂)_n-(δ1.34) and glycoprotein(δ2.02);The extract of the Electro-acupuncture group compared with that of the Model group has higher level of (CH₂)_n-(δ1.26,δ1.3),but lower level of choline(δ3.22).Among them,the molecules (CH₂)_n-were mainly from methylene compounds and fatty acid chain protons.3.the result of ¹H NMR spectra from kidneyThe result of the water soluble extract showed the obvious seperation among the three groups.The major chemical shifts which caused this seperation are at:δ1.27(lipid triglyceride),δ1.31-δ1.35(lactate),δ3.19-δ3.23(choline).The extract of the Model group compared with that of the Normal group has lower level of lactate (δ1.31-δ1.35);The extract of the Electro-acupuncture group compared with that of the Model group has higher level of lipid triglyceride(δ1.27),but lower level of choline (δ3.23).The result of the fat soluble extract:the signals among the three groups have a certain tendency of seperation,and the seperation between the Normal group and the rest of the two groups is more obvious.The major chemical shifts which contributed to the seperation are:δ1.26-δ1.34((CH₂)n-),δ2.38(-CH₂CO), δ2.86(C=CCH₂C=C),δ3.22(PC),δ5.38(CH=CH).The Model group showed significantly increased level of C=CCH₂C=C(δ2.86),CH=CH(δ5.38) and PC(δ3.22),significantly decreased level of CH₂) n-(δ1.3-δ1.34),-CH₂CO(δ2.38) when compared with the Normal group;The Electro-acupuncture group showed significantly increased level of(CH₂)n-(δ1.26),but decreased level of C=CCH₂C=C(δ2.86),CH=CH(δ5.38) and PC(δ3.22)) when compared with the Model group.Conclusion:1.The metabolic spectra of SAMP8 mice's blood plasma,hepatic tissue and renal tissue revealed a significant alteration when compared with those of SAMR1 mice. The major alteration in the spectra of SAMP8 mice showed mainly at these aspects:a.A reduced level of fucose(δ1.3),3-HB(δ1.22),and CH₃-(δ0.87—δ0.91) in the spectra of blood plasma;b.An increased glucose(δ3.79-δ3.87) and choline(δ3.22)levels but decreased taurine(δ3.27,δ3.43-δ3.47),glycoprotein(δ2.02) and(CH₂)_n-(δ1.34) levels in the spectra of hepatic tissue.c.An increased levels of PC(δ3.22) and unsaturated fatty acid(C=CCH₂C=C,δ2.86;CH=CH,δ5.38) but a decreased levels of lactate and saturated fatty acid9((CH₂)n-,δ1.3-δ1.34;-CH₂CO,δ2.38) in the spectra of renal tissue.2.The electro-acupuncture can influence the metabolic spectra of the SAMP mice's blood plasma,hepatic and renal tissue by adjusting the levels of specific molecules on the metabolic spectra.This influence is mainly on CH₃-(δ0.87—δ0.91) to the spectra of the blood plasma and on glucose(δ3.79-δ3.87) to that of the hepatic tissue as well as on PC(δ3.22) and unsaturated C=CCH₂C=C(δ2.86),CH=CH(δ5.38) to that of the renal tissue.