Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Zhengzhou University, No. 2, Jingba Road, Jinshui District, Zhengzhou, 450014, Henan Province, China.
Reprod Sci. 2022 Feb;29(2):564-577. doi: 10.1007/s43032-021-00719-8. Epub 2021 Sep 28.
Heat shock protein alpha 8 (HSPA8) was found to be downregulated in the placentas of patients with hypertensive disorders in pregnancy (HDP). We aim to explore the underlying role and mechanism of HSPA8 in HDP progression. Herein, HSPA8 mRNA expression in placentas and peripheral blood of patients with HDP and normal pregnant controls was measured with RT-qPCR. We found that HSPA8 expression was downregulated in placentas and peripheral blood of patients with HDP. HTR8/SVneo human trophoblast cells were transfected with pcDNA-HSPA8 or si-HSPA8. HSPA8 overexpression promoted cell proliferation, migration, and MMP-2 and MMP-9 protein levels, and inhibited apoptosis, while HSPA8 silencing showed the opposite results. Co-immunoprecipitation assay validated the binding between HSPA8 and β-arrestin1, as well as β-arrestin1 and A1AR proteins. HSPA8 bound with β-arrestin1 protein and promoted β-arrestin1 expression. β-arrestin1 bound with A1AR protein and inhibited A1AR expression. Then, HTR8/SVneo cells were transfected with pcDNA-HSPA8 alone or together with si-β-arrestin1, as well as transfected with pcDNA-β-arrestin1 alone or together with pcDNA-A1AR. β-arrestin1 silencing reversed the effects of HSPA8 overexpression on HTR8/SVneo cell functions. β-arrestin1 overexpression promoted cell proliferation migration, and MMP-2 and MMP-9 protein levels, and inhibited apoptosis, while these effects were reversed by A1AR overexpression. Lentivirus HSPA8 overexpression vector (Lv-HSPA8) was injected into a preeclampsia (PE) rat model, which attenuated blood pressure and fetal detrimental changes in PE rats. In conclusion, HSPA8 promoted proliferation and migration and inhibited apoptosis in trophoblast cells, and attenuated the symptoms of PE rats by modulating the β-arrestin1/A1AR axis. Our study provided a novel theoretical evidence and potential strategy for HDP treatment.
热休克蛋白 alpha 8（HSPA8）在妊娠高血压疾病（HDP）患者的胎盘组织中表达下调。本研究旨在探讨 HSPA8 在 HDP 进展中的潜在作用和机制。采用 RT-qPCR 检测 HDP 患者和正常妊娠对照者胎盘组织和外周血中 HSPA8 mRNA 的表达。结果发现，HSPA8 在 HDP 患者的胎盘组织和外周血中表达下调。用 pcDNA-HSPA8 或 si-HSPA8 转染 HTR8/SVneo 人滋养层细胞，结果发现 HSPA8 过表达促进细胞增殖、迁移和 MMP-2、MMP-9 蛋白水平，抑制细胞凋亡，而 HSPA8 沉默则呈现相反的结果。免疫共沉淀实验验证了 HSPA8 与β-arrestin1 以及β-arrestin1 与 A1AR 蛋白之间的结合。HSPA8 与β-arrestin1 蛋白结合并促进其表达。β-arrestin1 与 A1AR 蛋白结合并抑制其表达。然后，单独转染 pcDNA-HSPA8 或与 si-β-arrestin1 共转染，单独转染 pcDNA-β-arrestin1 或与 pcDNA-A1AR 共转染 HTR8/SVneo 细胞。β-arrestin1 沉默逆转了 HSPA8 过表达对 HTR8/SVneo 细胞功能的影响。β-arrestin1 过表达促进细胞增殖、迁移和 MMP-2、MMP-9 蛋白水平，抑制细胞凋亡，而 A1AR 过表达则逆转了这些作用。将慢病毒 HSPA8 过表达载体（Lv-HSPA8）注入子痫前期（PE）大鼠模型中，减轻了 PE 大鼠的血压和胎儿损害变化。综上所述，HSPA8 通过调节β-arrestin1/A1AR 轴促进滋养层细胞增殖、迁移和抑制凋亡，减轻 PE 大鼠的症状。本研究为 HDP 的治疗提供了新的理论依据和潜在策略。
