高糖条件下HRCECs中GALNT2与p-EGFR相关性研究

A Study on the Correlation between GALNT2 in HRCECs and p-EGFR under High Sugar Conditions

摘要: 目的：探讨在高糖条件下敲低多肽N-乙酰半乳糖胺转移酶2 (GALNT2)与抑制磷酸化表皮生长因子受体(Phosphorylated epidermal growth factor receptor, p-EGFR)对视网膜血管内皮细胞(HRCECs)增生及凋亡的影响。方法：本研究以高糖(25 mmol·L⁻¹)状态下体外培养的HRCECs为研究对象，将HRCECs分为mock组(25 mmol·L⁻¹葡萄糖)、mock + shGALNT2组(25 mmol·L⁻¹葡萄糖 + GALNT2敲低病毒)、mock + NC-shGALNT2 + Chrysophanol组(25 mmol·L⁻¹葡萄糖 + 对照病毒 + Chrysophanol)、mock + shGALNT2 + Chrysophanol组(25 mmol·L⁻¹葡萄糖 + GALNT2敲低病毒 + Chrysophanol)，连同培养液在37℃恒温培养箱中培养24 h，比较各组GALNT2 mRNA相对表达量、细胞增生值、细胞凋亡率，以及GALNT2、EGFR、EGF、p-EGFR蛋白相对表达量。结果：mock组、mock + shGALNT2、mock + NC-shGALNT2 + Chrysophanol组和mock + shGALNT2 + Chrysophanol组细胞中GALNT2 mRNA相对表达量比较，差异有统计学意义(P < 0.001)。mock + NC-shGALNT2 + Chrysophanol组和mock + shGALNT2 + Chrysophanol组中细胞增生值低于mock + shGALNT2组，差异具有统计学意义(P < 0.001)；mock + shGALNT2 + Chrysophanol组中细胞增生值高于mock + NC-shGALNT2 + Chrysophanol组，差异具有统计学意义(P < 0.001)。mock组、mock + shGALNT2组、mock + NC-shGALNT2 + Chrysophanol组和mock + shGALNT2 + Chrysophanol组细胞凋亡率分别为(8.66 ± 0.22)%、(5.47 ± 0.16)%、(11.94 ± 0.85)%和(6.70 ± 0.26)%，总体比较差异有统计学意义(F = 96.56, P < 0.001)。mock + shGALNT2 + chrysophanol组细胞凋亡率高于mock + shGALNT2组，差异有统计学意义(P < 0.001)；mock + shGALNT2 + chrysophanol组细胞凋亡率低于mock + NC-shGALNT2 + chrysophanol组，差异有统计学意义(P < 0.001)。mock + NC-shGALNT2 + Chrysophanol 组和mock + shGALNT2 + Chrysophanol组GALNT2蛋白相对表达量高于mock + shGALNT2组，差异有统计学意义(P < 0.001)。mock + NC-shGALNT2 + Chrysophanol组EGF、EGFR、p-EGFR蛋白相对表达量高于mock + shGALNT2组，但差异无统计学意义(P > 0.05)。结论：在高糖条件下，敲低的GALNT2可以通过介导p-EGFR可以提高HRCECs细胞的增殖能力，减少HRCECs细胞的凋亡。

Abstract: Objective: To investigate the effects of knocking down peptide N-acetylgalactosamine transferase 2 (GALNT2) and inhibiting phosphorylation of epidermal growth factor receptor under high glucose conditions on the proliferation and apoptosis of retinal vascular endothelial cells (HRCECs). Method: This study focuses on cultured HRCECs in vitro under high glucose (25 mmol·L⁻¹) conditions, and divides them into a mock group (25 mmol·L⁻¹ glucose), mock + shGALNT2 group (25 mmol·L⁻¹ glucose + GALNT2 knockdown virus), mock + NC-shGALNT2 + Chrysophanol group (25 mmol·L⁻¹ glucose + control virus + Chrysophanol), mock + shGALNT2 + Chrysophanol group (25 mmol·L⁻¹ glucose + GALNT2 knockdown virus + Chrysophanol) and culture medium were cultured in a constant temperature incubator at 37˚C for 24 hours. The relative expression level of GALNT2 mRNA, cell proliferation value, apoptosis rate, GALNT2, EGFR, EGF, and p-EGFR proteins were compared between the groups. Results: The relative expression levels of GALNT2 mRNA in the cells of the mock group, mock + shGALNT2 group, mock + NC-shGALNT2 + Chrysophanol group, were compared with mock + shGALNT2 + Chrysophanol group, and the differences were statistically significant (P < 0.001). Compared with mock + shGALNT2 group, the cell proliferation values in mock + NC-shGALNT2 + Chrysophanol and mock + shGALNT2 + Chrysophanol group were significantly reduced, and the differences were statistically significant (P < 0.001); Compared with mock + NC-shGALNT2 + Chrysophanol group, the cell proliferation value in mock + shGALNT2 + Chrysophanol group significantly increased, and the difference was statistically significant (P < 0.001). The apoptosis rates of mock group, mock + shGALNT2 group, mock + NC-shGALNT2 + Chrysophanol group and mock + shGALNT2 + Chrysophanol group were (8.66 ± 0.22)%, (5.47 ± 0.16)%, (11.94 ± 0.85)% and (6.70 ± 0.26)% respectively. The overall differences were statistically significant (F = 96.56), (P < 0.001). Compared with mock + shGALNT2 group, the apoptosis rate of the mock + shGALNT2 + Chrysophanol group was significantly increased, and the difference was statistically significant (P < 0.001); Compared with the mock + NC-shGALNT2 + Chrysophanol group, the apoptosis rate of the mock + shGALNT2 + Chrysophanol group was significantly reduced, and the difference was statistically significant (P < 0.001). Compared with mock + shGALNT2 group, the relative expression level of GALNT2 protein in the mock + NC-shGALNT2 + Chrysophanol and mock + shGALNT2 + Chrysophanol group significantly was increased, and the difference was statistically significant (P < 0.001). Comparison of relative protein expression levels of EGF, EGFR, and p-EGFR proteins in mock + shGALNT2, the mock + NC-shGALNT2 + Chrysophanol group showed an increasing trend, and the differences were not statistically significant (P > 0.05). Conclusion: Under high glucose conditions, knocking down GALNT2 can enhance the proliferation ability of HRCEC cells and reduce apoptosis by mediating p-EGFR.