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目的:从鲨鱼肝脏中分离纯化得到了一种鲨鱼肝刺激多肽(sHSP)。方法:将天然健康的条纹斑竹鲨鱼肝脏经过粗提,超滤,柱层析得到sHSP。通过采用MTT法,利用人肝癌SMMC-7721细胞和STZ损伤的小鼠胰岛β细胞瘤细胞来检测其生物活性。结果:通过MALDI-TOF-MS检测,sHSP的分子量为4 899 .715 Da ,其紫外特征吸收波长为273.1 nm。通过MTT法检测sHSP的刺激肝细胞再生的生物活性,结果显示,sHSP剂量为100μg.mL-1时,刺激指数为3.85 ,并且具有热稳定性和耐酸碱性。对STZ损伤的NIT-1β细胞,sHSP具有保护细胞完整性和修复细胞形态的作用。结论:我们从鲨鱼肝脏中分离纯化得到了一种能够显著刺激人肝癌细胞SMMC-7721再生,并且可以部分对抗STZ导致的NIT-1β细胞毒性损伤的活性肽。
OBJECTIVE: To isolate and purify a shark hepatic stimulating polypeptide (sHSP) from shark liver. METHODS: The sHSP was obtained from the liver of healthy and healthy stripe shark sharks after crude extraction, ultrafiltration and column chromatography. MTT assay was used to detect the biological activity of human hepatocellular carcinoma SMMC-7721 cells and STZ-injured mouse islet beta-cell tumor cells. Results: The molecular weight of sHSP was 4 899.715 Da by MALDI-TOF-MS, and its ultraviolet characteristic absorption wavelength was 273.1 nm. The bioactivity of sHSP-stimulated hepatocyte regeneration was measured by MTT assay. The results showed that when the sHSP dose was 100 μg.mL-1, the stimulation index was 3.85, and it had thermal stability and acid and alkali resistance. In STZ-injured NIT-1β cells, sHSP has the effect of protecting cell integrity and repairing cell morphology. Conclusion: We isolated and purified from the shark liver to obtain an active peptide that can significantly stimulate the regeneration of human hepatocellular carcinoma cell SMMC-7721, and can partially counteract the damage of NIT-1β cytotoxicity caused by STZ.