猫爪草多糖对小鼠腹腔巨噬细胞活力的调节作用

Regulating Effects of Polysaccharide from Radix Ranunculi Ternati on the Cell Vitality of Murine Peritoneal Macrophages

此次研究旨在探讨猫爪草多糖对体外培养的正常状态下的原代小鼠腹腔巨噬细胞活性的调节作用,以及小鼠腹腔巨噬细胞在体外培养条件下的活力变化情况。以原代培养的小鼠腹腔巨噬细胞为研究对象,设对照组(加入100μL DMEM培养基)和实验组(分别加入25μg/mL, 50μg/mL, 100μg/mL, 200μg/mL,400μg/mL的猫爪草多糖),分别采用噻唑蓝(MTT)比色法、CCK-8法、乳酸脱氢酶释放法和中性红吞噬实验检测不同浓度的猫爪草多糖对体外培养的小鼠腹腔巨噬细胞活力的调节作用;同时设置24 h、36 h、48 h、60 h和72 h的不同培养时间,观察在体外培养条件下,小鼠腹腔巨噬细胞活力的变化情况。结果表明:与对照组相比,不同浓度的猫爪草多糖均能增强小鼠腹腔巨噬细胞的活力,且猫爪草多糖浓度在100~400μg/mL的细胞活力极显著增强(p<0.01)。此外,各处理组的巨噬细胞在体外培养24~72 h不更换培养液的条件下,48 h处活性最佳。体外培养条件下,一定浓度的猫爪草多糖可以激活小鼠腹腔巨噬细胞,通过猫爪草多糖激活巨噬细胞,可能是猫爪草发挥提升机体免疫力的作用机制之一。此外,体外培养的巨噬细胞虽能存活长达一个月,但仍有一个最佳活力时间。

The aim of this research was to explore the effect of polysaccharide from Radix Ranunculi Ternati(PRT) on the cell viability of the normal murine peritoneal macrophages in vitro, as well as the the change of cell viability of the murine peritoneal macrophages cultured in vitro. This study took mouse peritoneal macrophages of primary culture as research object, designed a control group(only added with DMEM) and experimental groups(added with different concentration of PRT such as 25 μg/m L, 50 μg/m L, 100 μg/mL, 200 μg/mL, 400 μg/mL). And MTT assay, CCK-8 assay, the secretion of lactate dehydrogenase(LDH) as well as the phagocytic activity of neutral red were used to test the regulation of PRT on the activity of macrophages and the change of activity of the research object at different culture time in vitro(24 h, 36 h, 48 h, 60 h, 72 h). The results showed that compared with control group, PRT with different concentration could promote vitality of MPM, and that the concentration groups between 100 μg/mL and 400 μg/mL significantly elevated cellular viability. In addition, the macrophages cultured for 48 h in vitro showed the best activity. These results collectively indicated that the function of enhancing the body immunity of PRT might be correlated to its activation of macrophages. Despite the macrophages cultured in vitro might survive for weeks, it still possessed a optimizing period of time for cell vitality.