School of Public Health, Dali University, Dali, Yunnan Province, China.
School of Agronomy and Biological Sciences, Dali University, Dali, Yunnan Province, China.
J Obstet Gynaecol Res. 2022 Jan;48(1):188-199. doi: 10.1111/jog.15087. Epub 2021 Nov 2.
To investigate the protective effect of small peptides from Periplaneta americana (SPPA) on cyclophosphamide (CP)-induced premature ovarian failure (POF) in mice. Silent mating type information regulation 2 homolog 1 (SIRT1) /tumor-associated protein 53 (p53) signaling pathway plays an important role in delaying POF. Hematopoietic progenitor cell antigen (CD34) reflects ovarian aging from the side. However, whether SPPA inhibits POF in mice by influencing the SIRT1/p53 pathway and CD34 expression remains to be studied.
Forty female Kun Ming (KM) mice were divided into four groups: a control group (normal saline, n = 10), POF model group (160 mg/kg CP, n = 10), SPPA low-dosage group (160 mg/kg CP + 100 mg/kg SPPA, n = 10), and SPPA high-dosage group (160 mg/kg CP + 200 mg/kg SPPA, n = 10). CP administration route is intraperitoneal injection, and SPPA administration route is intragastric. Eyeball enucleation blood samples and the ovaries of mice were collected by midline laparatomy and oopherectomy, and the malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), follicle-stimulating hormone (FSH), and anti-Müllerian hormone (AMH) concentrations were tested. Immunohistochemical tests for the expressions of SIRT1, p53, and CD34 were carried out. Finally, ovarian mRNA levels of SIRT1 and p53 were detected with real-time fluorescence quantification PCR (qRT-PCR).
A mouse model of POF was generated using 160 mg/kg of CP. Compared with POF group, we found that plasma NO, MDA, and FSH decreased, while AMH and SOD increased in the SPPA low-dose group. Compared with the POF group, the SPPA low- and high-dosage groups achieved significant growth in the number of primordial, primary, and total number of healthy follicles at all levels, but sharp reductions in the number of atretic follicles. In addition, we found downregulated protein and mRNA expression of SIRT1, and upregulated that of p53 were observed in ovarian tissues of treated mice with POF, in immunohistochemistry experiments and qPCR experiments. In contrast, high protein and mRNA expression of SIRT1, and low that of p53 were observed in SPPA treatment groups. And the results of CD34 protein expression were consistent with that of SIRT1.
In total, SPPA significantly inhibited POF caused by CP in mice via activation of the SIRT1/p53 signaling pathway in the mouse ovary.
研究美洲大蠊小分子肽（SPPA）对环磷酰胺（CP）诱导的小鼠卵巢早衰（POF）的保护作用。沉默信息调节因子 2 同源物 1（SIRT1）/肿瘤相关蛋白 53（p53）信号通路在延缓 POF 中起重要作用。造血祖细胞抗原（CD34）从侧面反映卵巢衰老。然而，SPPA 是否通过影响 SIRT1/p53 通路和 CD34 表达来抑制小鼠的 POF 仍有待研究。
将 40 只昆明（KM）雌性小鼠分为 4 组：对照组（生理盐水，n=10）、POF 模型组（CP 160mg/kg，n=10）、SPPA 低剂量组（CP 160mg/kg+SPPA 100mg/kg，n=10）和 SPPA 高剂量组（CP 160mg/kg+SPPA 200mg/kg，n=10）。CP 给药途径为腹腔注射，SPPA 给药途径为灌胃。通过中线剖腹术和卵巢切除术采集小鼠眼球血样和卵巢，检测丙二醛（MDA）、一氧化氮（NO）、超氧化物歧化酶（SOD）、卵泡刺激素（FSH）和抗苗勒管激素（AMH）浓度。进行 SIRT1、p53 和 CD34 表达的免疫组织化学检测。最后，用实时荧光定量 PCR（qRT-PCR）检测卵巢 SIRT1 和 p53 的 mRNA 水平。
用 160mg/kg CP 建立了 POF 小鼠模型。与 POF 组相比，我们发现 SPPA 低剂量组血浆 NO、MDA 和 FSH 降低，而 AMH 和 SOD 升高。与 POF 组相比，SPPA 低剂量和高剂量组在各级原始卵泡、初级卵泡和总卵泡数量上均有明显增加，但闭锁卵泡数量明显减少。此外，我们在卵巢组织免疫组化实验和 qPCR 实验中观察到 POF 治疗小鼠 SIRT1 蛋白和 mRNA 表达下调，p53 表达上调。相反，在 SPPA 治疗组中观察到 SIRT1 蛋白和 mRNA 表达升高，p53 表达降低。CD34 蛋白表达结果与 SIRT1 一致。
总之，SPPA 通过激活小鼠卵巢中的 SIRT1/p53 信号通路，显著抑制 CP 诱导的小鼠 POF。
