Study On Effect Of Deproteinized Extract Of Calf Blood On Energy Metabolism In Hypoxia Mice

Deproteinted extract from calf blood(DECB) is a mixture of small molecules with biological activity which comes from deproteinted fetal bovine serum, including low molecular peptide, nucleoside, amino acids and oligosaccharides and other organic components and inorganic salts. The drug can increase transporting and utilization of oxygen and glucose of hypoxia cells during anoxic period, promoting mitochondrial oxidative phosphorylation, improving energy metabolism of cells. The drug is widely used in clinical, but the specific mechanism, the target and metabolic pathway hasn’t been clear.Objective: Through the overall level and molecular level this article analysis energy metabolism mechanism of DECB, at the same time, using of gene chip technology to explore DECB target gene and draws the related metabolic pathway map. Studying on Mechanism of DECB will provide unlimited space for future clinical application.Methods: 1. Using soft ionization method and flight mass spectrometry to detect the molecular weight range of DECB and getting the respiratory activity Stimulation index(SI) value by Warburg apparatus. 2. Through the overall level experimental to study the energy metabolism mechanism of DECB. 24 ICR mice, male, body weight 18-22 g, randomly divide into four groups according to the weight, each group has 6 mice, that is the control group, the drug group, model group and drug model group, the control group and the model group were fed with distilled water for 5 weeks, drug group and drug model group was continuously injected of DECB for 5 weeks at a dose of 3.3mg/20 g subcutaneously. After 1h of the last injection is finished, the mice of model group and drug model group are respectively arranged in the grinding port glass bottle, sealed. Severe hypoxia can cause convulsions and loss of reflexes, recorded the time of tolerance to hypoxia. Picking eyeball to take blood, take the heart, brain, liver and other tissues; Determination glucose content of serum, heart, brain and liver; Determination ATP content of heart, brain and liver; Determination lactate content of blood, liver and brain, as well as the determination protein content of brain, heart and liver. 3. Take the mice liver of blank group and the drug group to extract m RNA, and making high flux gene chip. Analysis of the differentially expressed genes of the blank group and the drug group, and through gene function annotation and gene enrichment analysis screen of functional genes related to energy metabolism, through the analysis of literature and gene annotation we choose 13 genes of metabolic pathway of glucose and lipid for real-time fluorescence Real Time PCR validation.Results: 1.The results showed that the molecular weight distribution of DECB was <3000D; the respiratory activity value is: QO2(blank) =2.03, QO2(sample) =5.25, SI=2.59.2. Biochemical test results show that: compared with the blank model group, the drug model group significantly prolonged hypoxia tolerance time(P﹤0.05); Compared with the blank group, and the content of glucose in the drug group was significantly lower than that in the control group(P< 0.05), and compared with the blank model group, the blood glucose level of the model mice was significantly reduced(P< 0.01), compared with the blank group, DECB can make the glucose content of heart and liver significantly increased(P<0.05), Compared with the blank model group, DECB can significantly improve glucose content mouse of brain(P<0.01), heart(P<0.05) and liver(P<0.01); DECB can significantly(P<0.01) reduced the amount of lactic acid in the blood, brain and liver of Hypoxia mice. DECB can significantly(P<0.01) increase ATP content of brain, heart and liver of hypoxia mice. At physiological state, DECB can significantly increase protein content of brain(P<0.05), heart(P<0.01) and liver(P<0.01), and at the pathological state DECB can also significantly increase protein content of heart(P<0.01) and liver(P<0.01).3. The results of energy metabolism gene expression profile show that there were some differences in genes.Conclusion: DECB is of high biological activity which can prolong hypoxia tolerance time of hypoxia mouse, improve energy metabolism, reduce the lactic acid content of hypoxia in the brains of mice, could improve glucose utilization rate and increase the rate of ATP synthesis. The mechanism is mainly affected by several key genes regulating like Hngcs2, Cpt1 a, Angptl4, Cyp8b1 and Ehhadh.