Study On Fatigue Resistance And Radiation Damage Resistance Of Two Typical Macamides

Macamides were special kinds of amide active compound in Maca and it has attracted more and more attention due to its various biological activities.However,because of its variety,low content and difficulty in isolation and extraction of monomers from natural sources,the pharmacodynamic study of macamide was limited to the overall activity evaluation and pharmacodynamic evaluation of extracts and components,and the pharmacological target is still unknown.Therefore,a high-efficiency,convenient and systematic synthesis and preparation process of macamide monomer was established,and the biological activities of macamide monomers with different structural characteristics were investigated,and further explore the structure source and molecular law of the difference in activity.It can support clear technical support and theoretical basis for the healthy development of Maca industry and the pharmacological efficacy of macamide.Based on this,this project intends to establish a method of synthesizing macamide by using carbodiimide condensing agent method,and combining animal experiments in vivo to evaluate the effects of different macamide monomers on relieving physical fatigue and anti-radiation damage.By comparing N-benzylhexadecanamide(NBH)of high saturation,N-benzyldocosahexaenoicamide(NBDHA)of high unsaturation and ethyl esterified docosahexaenoicacid(DHA-EE),the characteristics and laws between the structure and biological activity of Macamide monomers were discussed.Objective:To develop the synthesis and preparation technology of compounds with different saturation and typical structure characteristics of Macamide,and to evaluate the biological activity of the prepared Macamide monomer and explore its source of activity by establishing animal models of fatigue and radiation damage in mice.Methods: 1.Synthesis and structure analysis of macamide The macamide and its similar compounds are synthesized by a carbodiimide condensing agent method,a saponification reaction,a low-temperature crystallization,a semi-preparative liquid phase purification compound,and a high-performance liquid phase(HPLC),infrared(IR,liquid chromatography-mass spectrometry(LC-MS)and nuclear magnetic resonance(NMR)techniques were used to identify compound structures.2.Macamide relieves physical fatigue The physical fatigue model of mice was set up,then the weight change and weight-bearing swimming time of mice were measured,the related biological indexes of mice were tested,the activities of LDH and CK in serum,the contents of NEFA and BA,the changes of SOD and MDA,and the pathological changes of related tissues were detected.3.Macamide improves radiation damage in mice The model of radiation injury in mice was set up,and the appropriate radiation dose was searched.After irradiation,the mice were continuously intragastric administration for 7 days.Then the weight change,peripheral blood cell count in blood,spleen index and pathological condition,the changes of SOD and MDA in serum,bone pathological condition,apoptosis of bone marrow cells and proliferation of surface stem cells were detected by flow cytometry,and the expression of related proliferative genes were detected by QT-PCR.Results: 1.Synthesis and structure analysis of macamide Two kinds of macamide compounds were successfully synthesized by a carbodiimide condensing agent method,N-benzylhexadecanamide was a white powder,and N-benzyldocosahexaenoicacidamide was a pale yellow oil.After purification,the purity of the two compounds was >95% or more by high performance liquid phase,and the yield was >30%.The compounds were identified by IR,LC/MS and NMR as N-benzylhexadecanamide and N-benzyldocosahexaenoicacidamide.2.Macamide relieves physical fatigue Both macamide compounds could prolong the weight-loading swimming time of mice.The results of various biological indicators showed that two kinds of macamide compounds can reduce the levels of BLA and BUN,LDH and CK;increase the levels of LG and MG;reduce the serum MDA content and increase the activity of SOD;histological analysis shows that two kinds of horses The carbamide compound has protective effects on mouse skeletal muscle and cardiomyocytes and liver during exercise.3.Macamide improves radiation damage in mice The dose of 6.0 Gy was selected as the median lethal dose;the weight loss of mice in each drug-administered group was relieved;each drug-administered group increased the number of white blood cells,red blood cells,hemoglobin and platelets in the blood;increased the activity of SOD in serum and decreased The level of MDA increased;the spleen index increased,the spleen lesions were alleviated,the number of nucleus in the bone increased,and the vacuolization decreased;the number of bone marrow cells increased,the apoptosis decreased,and the proportion of stem cells increased in each administration group;Each group may mediate the proliferation of bone marrow cells through NRF-2,IL-3,P53,CyclinD1 gene,thereby restoring hematopoietic function.Conclusion:Based on the method of carbodiimide condensation agent,two typical macamide compounds were successfully prepared: N-benzylhexadecanamide with the highest content in natural Maca and N-benzyldocosahexaenoicacidamide with the highest potential unsaturation yet to be found in natural maca.Among them,the purity of N-benzylhexadecanamide is 98%,the yield is 39.15%,and the purity of N-benzyldocosahexaenoicacidamide is more than 95%,the yield is 30.12%.Based on fatigue mice model and irradiation injury mice model,the effects of N-benzylhexadecanami-de,N-benzyldocosahexaenoicacidamide and ethyl esterified docosahexaenoicacid on alleviating physical fatigue and radiation injury were investigated.The results showed that the three compounds had significant alleviating effects on fatigue mice and irradiated mice.The overall improvement of N-benzyldocosahexaenoicacidamide and ethyl esterified docosahexaenoicacid with high unsaturation degree was better than that of N-benzylhexadecanoicacidamide with high saturation degree.The improvement effect of N-benzyldocosahexaenoicacidamide and ethyl esterified docosahexaenoicacid was similar.It is concluded that the main source of the activity of macamides is the fat chain,and there is a certain correlation between the anti-fatigue and anti-radiation damage activities and the unsaturation of the fat chain.