解剖科学进展

目的 探究黄蜀葵花制剂对与宫颈癌细胞共培养的巨噬细胞M2极化的影响及机制。方法 THP-1细胞经PMA诱导为M0型巨噬细胞，将其与SiHa细胞共培养，共培养的细胞分为对照组(Control组)、黄蜀葵花制剂低剂量组(AM-L组)和黄蜀葵花制剂高剂量组(AM-H组),加入黄蜀葵花制剂后收集巨噬细胞及细胞培养液上清，流式细胞术检测细胞中CD11b+CD206+细胞比例，ELISA方法检测细胞培养液上清中IL-10和IL-4水平，Western blot检测巨噬细胞中p-FAK和p-ERK2蛋白表达。用黄蜀葵花制剂处理的SiHa细胞共培养的巨噬细胞的条件培养液培养SiHa细胞，CCK-8法检测细胞增殖活性，流式细胞术检测细胞凋亡率。结果 黄蜀葵花制剂处理的与宫颈癌细胞共培养的巨噬细胞中CD11b+CD206+细胞比例明显降低，且细胞培养上清液中IL-10和IL-4水平同样明显降低。用黄蜀葵花制剂处理的宫颈癌细胞共培养的巨噬细胞的条件培养液培养的宫颈癌细胞的增殖活性以及迁移和侵袭能力明显降低，细胞的凋亡率明显升高。黄蜀葵花制剂处理的与宫颈癌细胞共培养的巨噬细胞中p-FAK和p-ERK2蛋白表达水平明显降低。结论 黄蜀葵花制剂通过抑制宫颈癌免疫微环境中M2型巨噬细胞极化抑制宫颈癌发展，其机制与抑制巨噬细胞中FAK/ERK2信号通路激活有关。

Objective To explore the effect and mechanism of abelmoschus manihot preparation on M2 polarization of macrophages co-cultured with cervical cancer cells.Methods THP-1 cells were induced into M0-type macrophages by PMA and co-cultured with SiHa cells.The co-cultured cells were divided into control group, low-dose of abelmoschus manihot preparation group(AM-L group)and high-dose of abelmoschus manihot preparation group(AM-H group).After adding abelmoschus manihot preparation,macrophages and cell culture supernatant were collected, the proportion of CD11b+ CD206+ cells in cells was detected by flow cytometry, the levels of IL-10 and IL-4 in cell culture supernatant were detected by ELISA,and the expressions of p-FAK and p-ERK2 proteins in macrophages were detected by Western blot.SiHa cells were cultured in the conditioned medium of macrophages co-cultured with SiHa cells treated with Hibiscus manihot flower preparations.The proliferation activity of the cells was detected by CCK-8 method, and the apoptosis rate was detected by flow cytometry.Results The ratio of CD11b+ CD206+ cells in macrophages co-cultured with cervical cancer cells treated with abelmoschus manihot preparation were decreased significantly, and the levels of IL-10 and IL-4 in cell culture supernatant also were decreased significantly.The proliferation activity, migration and invasion ability of cervical cancer cells cultured in the conditioned medium of macrophages co-cultured with the preparation of abelmoschus manihot were decreased obviously, and the apoptosis rate of cells was increased obviously.The expressions of p-FAK and p-ERK2 proteins in macrophages co-cultured with cervical cancer cells treated with abelmoschus manihot preparation were decreased significantly.Conclusion Abelmoschus manihot preparation inhibits the development of cervical cancer by inhibiting the polarization of M2 macrophages in the immune microenvironment of cervical cancer, and its mechanism is related to inhibiting the activation of FAK/ERK2 signaling pathway in macrophages.