RESUMEN
ObjectiveProteomics was used to investigate the protein differences between porcine cardiac blood（PCB） and porcine blood（PB） from Menghe medical school and to compare the effects of both on the microglial inflammation of Salviae Miltiorrhizae Radix et Rhizoma（DS）. MethodNanoliquid chromatography-quadrupole orbitrap mass spectrometry（nLC-MS/MS） and bioinformatics were utilized to compare the proteomic differences of PCB and PB in simulated gastrointestinal digestion. Furthermore， Western blot was used to verify the contents of some shared proteins and differential proteins identified in PCB and PB. In addition， BV2 neuroinflammation model constructed by corticosterone（CORT） and lipopolysaccharide（LPS） was applied to detect the intervention effects of PCB and PB on the levels of tumor necrosis factor（TNF）-α and interleukin（IL）-6 in BV2 inflammatory cells of DS. ResultA total of 69 common proteins and 68 differential proteins were identified in PCB and PB， among which the common proteins included transferrin（Tf） with brain-targeting effect， and the differential proteins in the two were 41 and 27， respectively. Western blot validation showed that the difference in the content of the same protein Tf between PCB and PB was not statistically significant， while the difference in the contents of the specific proteins of creatine kinase M and heart-type fatty acid binding protein（H-FABP） were statistically significant（P<0.05）. Moreover， in vitro experimental studies revealed that compared with the same concentration of DS group， in addition to the 100 mg·L-1 PB-DS group， PCB-DS and PB-DS groups could significantly inhibit the levels of TNF-α and IL-6 in BV2 inflammatory cells（P<0.05， P<0.01）， and PCB-DS group had more significant anti-inflammatory effect than PB-DS group with the same concentration（P<0.05， P<0.01）. ConclusionBoth of PCB and PB can enhance the inhibitory effect of DS on the release of inflammatory factors， thus playing a neuroprotective role， and PCB promotes DS inhibition more significantly， which may be due to the existence of the two involved in energy metabolism-related differential proteins， which can lay a foundation for revealing the scientific connotation of the processing of PCB-DS and PB-DS.
RESUMEN
OBJECTIVE: To establish and optimize the preparation method of protoporphyrin disodium (NAPP) from non-anticoagulant porcine blood.METHODS: Heme was prepared from non-anticoagulant porcine blood firstly.Ferrous sulfide was added into heme to prepare NAPP by saponification and esterification reaction.UV absorption spectrum and infrared spectrum were applied for qualitative test,and ultraviolet spectrophotometry was adopted for quantitative detection of NAPP.Orthogonal experiment was used to optimize the preparation method of NAPP and to determine the optimal parameters of the preparation process.RESULTS: The preparation method of NAPP from non-anticoagulant porcine blood was established.10.6 g heme could be obtained from per kilogram of non-anticoagulant porcine blood and 3.46 g NAPP could be obtained from per 5 g heme.The purity quotient of NAPP was 88.8%.The optimal parameters for the preparation of NAPP were as followed: incremental amount of methanol vs.chloroform was 70 mL ∶ 120 mL in estrification process;Incremental amount of toluene vs.1 mol?L-1 sodium hydroxide-methanol solution was 50 mL ∶ 80 mL in saponification process.The sample was extracted by continuous therma reflux at 90 ℃ for 2.0 h.CONCLUSION: The established preparation method of NAPP from non-anticoagulant porcine blood is an ideal approach for the preparation of NAPP with simple operation,abundant raw material,short preparation cycle,low cost and high yield.