Studies On The Anti-fatigue And Hematopoiesis Enhancement Effects Of Calf Spleen Extractive Injection

Cancer,also called malignancy,involvs more than 100 kinds of diseases.There are five people die of cancer per minute in the world,and cancer patients usually suffer from fatigue.Currently,chemotherapy and radiotherapy are still the main treatments of malignancies,but the suppression of blood cells and hematopoietic tissue seriously limits their effectiveness and applications.Therefore,it is of great significance to find therapeutic agents that possess protective effect on hematopoietic function and are beneficial for the recovery of patients after chemotherapy and radiotherapy.Calf Spleen Extractive Injection(CSEI),extracted from the spleen of healthy cows(within 24 hours of birth),is a small-peptide-enriched extraction.The pharmacological foundation of CSEI is based on the activation of non-specific immunity,promotion of bone marrow stem cells and the enhancement of hematopoietic function.The clinical analysis shows that about 40%of CSEI is used in the treatment of cancer.CSEI can reduce the bone marrow suppression,enhance the patient’s strength,and improve the patient’s life quality.In this study,variety of biological methods were used to evaluate the anti-fatigue effect of CSEI and its promotion effect on hematopoiesis and the underlying mechanism.The main contents are listed as follows:1.Study on the anti-fatigue effect of CSEIThe Balb/c mice were used in this experiment.Exercise tolerance test,fatigue and oxidation related biochemical indices and plasma hormone level were detected after 21 days’intraperitoneal administration.The results showed that CSEI significantly improved the running and loading swimming time of mice,as well as the rolling stick time.The measurement of LD,LDH and glycogen showed that CSEI could simultaneously reduce the LD(in plasma,liver and muscle)and LDH(in liver and muscle)levels,and increase the liver glycogen and muscle glycogen.Meanwhile,CSEI improved the SOD,GSH-Px levels in plasma,liver and muscle,decreased ROS level both in liver and muscle,and the variation of plasma hormone testosterone,estradiol and cortisol was also viewed.In conclusion,CSEI displayed an anti-fatigue effect possibly through antioxidant and hormone regulation.2.Study on the hematopoietic function of CSEI in vitroIn order to primarily evaluate the hematopoietic function of CSEI,K562 and CHRF cells were used as the hematopoietic cells model.Cell viability,cell apoptosis,cell differentiation,and hematopoietic function-related proteins expression were studied.The results showed that CSEI could promote the proliferation of K562 and CHRF cells but couldn’t induce the apoptosis of these two types of cells;CSEI induced the differentiation of K562 into erythrocytes;CSEI activated hematopoietic-and proliferation-related factors RSK1p90,ELK1 and c-Myc,and facilitated the expression of differentiation-and maturation-related transcription factors GATA-1 and GATA-2,indicating that the role of CSEI in promoting the function of hematopoietic cells may be due to the regulation of genes by transcription factors.3.Study on the protective effect of CSEI on hematopoietic injury mice and its undering mechanismIn order to further determine the hematopoietic function of CSEI,cyclophosphamide (CTX)was used to establish the hematopoietic dysfunction mice model.The effect of CSEI on hematopoietic function and its possible mechanism were studied in vivo,and then the possible mechanism was further verified in primary cultured bone marrow mononuclear cells(BMNCs).In the mice with hematopoietic suppression,3 weeks of CSEI administration enhanced the bodyweights and thymus indices,suppressed the spleen indices and strongly elevated the production of HSC/HPCs,neutrophils and B cells in bone marrow,ameliorated bone marrow cellularity,and regulated the ratio of peripheral blood cells.Proteome profiling combined with ELISA revealed that CSEI regulated the levels of cytokines,especially G-CSF and its related factors,in the spleen and plasma.Additional data revealed that CSEI promoted phosphorylation of STAT3,which was stimulated by G-CSF in both mice spleen and cultured BMNCs.Taken together,CSEI has the potential to improve hematopoietic function via the G-CSF-mediated JAK2/STAT3 signaling pathway.In summary,CSEI has the anti-fatigue effect and is a safe and effective drug for promoting hematopoiesis.It promotes hematopoietic function through G-CSF-mediated JAK2/STAT3 signaling pathway.This study provided the pharmacological basis for the clinical use of CSEI on the anti-fatigue and radiotherapy and chemotherapy of cancer and other symptoms induced by hematopoietic insufficiency.