姜黄素对人翼状胬肉成纤维细胞增殖和凋亡及迁移的影响

Effects of curcumin on proliferation and apoptosis and migration of human pterygium fibroblasts

摘要:

目的：探讨姜黄素对体外培养的人翼状胬肉成纤维(HPF)细胞增殖和凋亡及迁移的影响。

方法：收集我院2021-11-24/12-16手术切除的翼状胬肉组织7例，进行原代成纤维细胞体外培养，并采用免疫荧光染色法进行细胞鉴定。采用含等量二甲基亚砜的0、10、20、40、80、160μmol/L姜黄素处理HPF细胞24h后，用CCK8法检测细胞增殖情况。根据CCK8检测结果将细胞分为对照组(不含姜黄素)、20μmol/L姜黄素组、40μmol/L姜黄素组，每组分别处理HPF细胞24h。采用流式细胞仪检测细胞凋亡，Transwell迁移实验检测细胞迁移，实时荧光定量PCR法和Western blot法检测Bax、Bcl-2、Cyclin D1及MMP2的mRNA与蛋白表达。

结果：与对照组相比，20μmol/L姜黄素组和40μmol/L姜黄素组均能抑制HPF细胞的增殖和迁移，并诱导其凋亡(均P<0.05)。与对照组相比，20μmol/L姜黄素组能下调Cyclin D1及MMP2，上调Bax的mRNA表达，下调Bcl-2的蛋白表达(均P<0.05)。与对照组相比，40μmol/L姜黄素组能下调Bcl-2、Cyclin D1及MMP2的mRNA与蛋白表达，上调Bax的mRNA与蛋白表达(均P<0.05)。与20μmol/L姜黄素组相比，40μmol/L姜黄素组能下调MMP2的mRNA表达，下调Bcl-2的蛋白表达，上调Bax的mRNA与蛋白表达(均P< 0.05)。

结论：姜黄素可通过抑制Cyclin D1的表达抑制HPF细胞的增殖，通过下调Bcl-2并上调Bax的表达来诱导HPF细胞的凋亡，通过下调MMP2的表达来抑制HPF细胞的迁移。

Abstract:

AIM: To investigate the effects of curcumin on the proliferation and apoptosis and migration of human pterygium fibroblasts(HPF)in vitro.

METHODS: A total of 7 cases of pterygium tissue removed at our hospital from November 24, 2021 to December 16, 2021 were collected. Then, primary fibroblasts were cultured in vitro and identified by immunofluorescence staining. HPF were treated with 0, 10, 20, 40, 80 and 160μmol/L curcumin containing equal amount of dimethyl sulfoxide for 24h, then the cell proliferation was detected by CCK8 assay. According to the results of CCK8, the cells were divided into control group, 20μmol/L curcumin group and 40μmol/L curcumin group, and the cells were treated with corresponding concentration of curcumin for 24h in each group. Flow cytometry was used to detect apoptosis, Transwell migration assay was used to detect cell migration, and real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the expression of mRNA and protein of B-cell lymphoma-2 associated X protein(Bax), B-cell lymphoma-2(Bcl-2), Cyclin D1 and matrix metalloproteinase 2(MMP2).

RESULTS: Compared with the control group, both 20μmol/L curcumin group and 40μmol/L curcumin group can inhibit the proliferation and migration of HPF and induce its apoptosis(all P<0.05). Compared with the control group, 20μmol/L curcumin group can down-regulate the mRNA expression of Cyclin D1 and MMP2, up-regulate the mRNA expression of Bax, and down-regulate the protein expression of Bcl-2(all P<0.05). Compared with the control group, 40μmol/L curcumin group can down-regulate the expression of mRNA and protein of Bcl-2, Cyclin D1 and MMP2, and up-regulate the expression of mRNA and protein of Bax(all P<0.05). Compared with 20 μmol/L curcumin group, the 40 μmol/L curcumin group can down-regulate the mRNA expression of MMP2, down-regulate the protein expression of Bcl-2, and up-regulate the mRNA and protein expression of Bax(all P<0.05).

CONCLUSION: Curcumin can inhibit the proliferation of HPF by inhibiting the expression of Cyclin D1, induce the apoptosis of HPF by down-regulating Bcl-2 and up-regulating the expression of Bax, and inhibit the migration of HPF by down-regulating the expression of MMP2.