红茴香根皮提取液对神经细胞生长和增殖作用的影响

Effect of Illicium angustisepalum A.C.Smith root bark extract on the growth and proliferation of nerve cells

目的探讨红茴香根皮提取液对神经细胞生长和增殖的影响。方法①对未分化的PC12细胞的促神经生长作用:将未分化的PC12细胞制成细胞悬液,分成8组:阴性对照组、神经生长因子(NGF)阳性对照组、红茴香根皮提取液Ⅰ、Ⅱ、Ⅲ组,以及红茴香根皮提取液Ⅰ、Ⅱ、Ⅲ分别与NGF(50μg/L)联合用药组。连续3 d(24、48、72 h)进行光镜观察,计算突起生长细胞(≥2倍细胞直径)占总细胞的百分率。②对NGF分化的PC12细胞的促增殖作用:取NGF诱导分化的PC12细胞,分成8组:阴性对照组、NGF阳性对照组、红茴香根皮提取液Ⅰ、Ⅱ、Ⅲ组、以及红茴香根皮提取液Ⅰ、Ⅱ、Ⅲ与NGF(300μg/L)联合用药组。用酶联免疫检测仪测定570 nm处的吸光度(OD_(570)),计算增殖率。结果①促神经生长作用:与阴性对照组相比,提取液各剂量组均能显著促进PC12细胞突起生长,第2天(48 h)诱导分化活性最强,提取液各剂量组与NGF联合应用时,对促进PC12细胞突起生长有明显的协同作用。②促增殖作用:在有血清培养条件下,红茴香根皮提取液中剂量和高剂量组均对PC12细胞有明显促增殖作用;在无血清培养条件下,受试药红茴香根皮提取液各剂量组均对PC12细胞有明显促增殖作用,与阴性对照组比差异均有统计学意义(P<0.05)。结论红茴香根皮提取液可能是神经营养因子样物质,具有促神经细胞生长和增殖作用。

Objective To investigate the effect of Illicium angnstisepalum A. C. Smith root bark extract on the growth and proliferation of nerve cells. Methods （ 1 ） Its effect on the nerve cell growth of undifferentiated PC12 cells： undifferentiated PC12 cells were prepared as cell suspension and was then divided into 8 groups and subgroups： the negative control group, the nerve growth factor （NGF） control group, the Illicium angustisepalum A. C. Smith root bark extract sub-groups I, II and III, and the subgroups I, II and III respectively combined with NGF （50 μg/L）. Then, microscopic observation was performed for 3 successive days （24,48,72 h） and the percentage of neurite outgrowth of PC12 cells in total cell growth was calculated. （2） Its effect on the proliferation of differ- entiated PC12 cells promoted by NGF： Differentiated PC12 cells promoted by NGF were collected and divided into 8 groups and sub- groups： the negative control group, the NGF control group, the Illicium angustisepalum A. C. Smith root bark extract sub-groups I, II and III, and the subgroups I, II and III respectively combined with NGF （300 μg/L）. Photoabsorption rate at 570 nm was determined with ELISA and proliferation rate was then also calculated. Results （ 1 ） The effect on the growth of nerve cells ： when compared with the negative control group, different doses of the abstract could all promote significantly the differentiation of PC12 cells, with the pro moting effect being the strongest on the second day （48 h）. When different doses of the abstract were combined with NGF, synergistic effect of the abstract on the growth of PC12 could be observed significantly. （2） The effect on the proliferation of nerve cells： In serum culture conditions, the moderate and high doses of the abstract all had obvious differentiation effect on PC12 cells. In serum - free cul ture conditions, different doses of the abstract also had obvious proliferation effect on PC12 cells, and statistical significance could benoted, when compared with the negative control group （ P 〈 0.05 ）. Conclusion Illicium angustisepalum A. C. Smith root bark extract might be a neurotrophic factor-like substance, which could promote the growth and proliferation of nerve cells.