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将本室自制的林蛙皮多肽进行SDS-PAGE和紫外扫描检测,并将其以不同质量浓度作用于人永生化表皮细胞(HaCaT细胞),利用MTS法、Annexin V-FITC/PI法分别检测林蛙皮多肽对HaCaT细胞增殖及其凋亡的影响。结果显示,林蛙皮多肽在20%SDS-PAGE条件下呈现弥散状态,灰度分析表明相对分子质量在10 000以下的多肽占总多肽的67.1%。林蛙皮多肽在(50~800)mg/L条件下,随着多肽质量浓度的升高,对HaCaT细胞促进增殖的作用逐渐增强,当林蛙皮多肽质量浓度为800mg/L时,MTS检测HaCaT细胞活力D值是正常对照细胞D值的2.5倍(P<0.01);此时,细胞凋亡率比正常对照细胞降低了(2.7±0.03)%,(P<0.01)。以上结果表明,林蛙皮活性多肽对HaCaT细胞的增殖有显著的促进作用,并对其凋亡有显著的抑制作用。
SDS-PAGE and UV-scan were used to detect the polypeptide of self-made Rana broomculonus in our laboratory. The peptides were applied to immortalized human epidermal cells (HaCaT cells) with different mass concentrations and detected by MTS and Annexin V-FITC / PI Effect of Rana broomcorn polypeptide on the proliferation and apoptosis of HaCaT cells. The results showed that the Rana spp skin polypeptide showed a diffuse state under the condition of 20% SDS-PAGE. The gray scale analysis showed that the polypeptide with a relative molecular mass of less than 10,000 was 67.1% of the total polypeptide. Under the condition of (50 ~ 800) mg / L, the activity of Rana broomculata polypeptide on HaCaT cells increased gradually with the increase of the mass concentration of polypeptide. When the polypeptide concentration of Rana sylvatica was 800mg / L, The cell viability D of HaCaT cells was 2.5 times higher than that of normal control cells (P <0.01). At this time, the apoptotic rate of HaCaT cells was decreased by (2.7 ± 0.03)% compared with that of normal cells (P <0.01). The above results show that Rana broomcorn active polypeptide can significantly promote the proliferation of HaCaT cells and significantly inhibit the apoptosis of HaCaT cells.