黑枸杞花青素通过Ⅲ-PI3K通路调控小鼠巨噬细胞RAW264.7自噬

Lycium ruthenicum murry anthocyanin Extracts Regulates Autophagy in Mouse Macrophage RAW264.7 viaⅢ-PI3K Pathway

为探讨青海柴达木黑枸杞花青素对小鼠巨噬细胞RAW264.7自噬的影响,用CCK-8法检测不同浓度不同时间黑枸杞花青素诱导小鼠巨噬细胞RAW264.7的增殖情况;用Western Blot检测不同浓度黑枸杞花青素浓度诱导小鼠巨噬细胞RAW264.7后自噬因子LC3-Ⅱ和Beclin-1的蛋白表达情况;用Western Blot检测Ⅲ-PI3K自噬调控通路中Ⅲ-PI3K和Beclin-1自噬因子及其结合3-MA自噬抑制剂后蛋白的表达情况。结果表明,25μg/mL的黑枸杞花青素诱导24 h小鼠巨噬细胞RAW264.7的细胞活力均显著增加(P<0.05),增殖效果最好;随着黑枸杞花青素诱导小鼠巨噬细胞RAW264.7浓度的提高,LC3-Ⅱ和Beclin-1自噬因子的蛋白表达显著降低(P<0.01);在3-MA作用后,花青素显著降低了Ⅲ-PI3K和Beclin-1的蛋白表达(P<0.05)。试验结果表明,黑枸杞花青素诱导小鼠巨噬细胞RAW264.7增殖的最佳浓度为25μg/m L,时间为24 h。同时黑枸杞花青素能抑制小鼠巨噬细胞RAW264.7自噬,其调控机理可能与Ⅲ-PI3K自噬调控通路有关。

To explore the effect of lycium ruthenicum murry anthocyanin extracts on the autophagy in mouse macrophage RAW264.7,the experiment used CCK-8 to measure the effect of lycium ruthenicum murry anthocyanin extracts of different concentrations and concanavalin protein(Con A)on proliferation in RAW264.7 at different time.Western blot was performed to detect the effect of lycium ruthenicum murry anthocyanin extracts of different concentrations on the protein expression of LC3-Ⅱand Beclin-1 in RAW264.7.Besides,further study on the protein expression of LC3-Ⅱand Beclin-1 induced by 3-MA inⅢ-PI3K pathway was detected by Western Blot.Results showed that 25μg/mL of lycium ruthenicum murry anthocyanin extracts significantly increased cell viability of RAW264.7 at 24 h(P<0.05);The protein expression of LC3-Ⅱand Beclin-1 was significantly reduced and showed dose-dependence(P<0.01)with the concentration;Ⅲ-PI3K and Beclin-1(P<0.05)was significantly reduced after the 3-MA effect.The results revealed the best concentration was 25μg/mL at 24 h.Meanwhile,lycium ruthenicum murry anthocyanin extracts could inhibit the autophagy of RAW264.7.In addition,the mechanism of autophagy may be related to the regulation ofⅢ-PI3K pathway.