The Role of Cryptotanshinone Regulates HTR-8/SVneo Cells Activity Through the p53/STAT3 Pathway in Unexplained Recurrent Spontaneous Abortion

IF 2.4 3区医学 Q3 IMMUNOLOGY

American Journal of Reproductive Immunology Pub Date : 2025-09-29 DOI:10.1111/aji.70163

Lidan Lu, Ximei Cai, Yang Zhang, Weibo Zhao, Haiyan Ni, Liping Zhang, Peijuan Wang

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Abstract

Objective

This study sought to investigate the therapeutic effects and mechanisms of cryptotanshinone (CT) on unexplained recurrent spontaneous abortion (URSA).

Methods

The DBA/2 male and CBA/J female mice were caged together to construct a URSA model before treatment with high and medium-low doses of CT. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of interleukin (IL)-1 beta (β), IL-17, and IL-22 in serum in each category of mice. Hematoxylin–eosin (HE) pathology was employed to detect decidua and uterine tissues, while polymerase chain reaction (PCR) was utilized to ascertain relative expression levels of serum messenger ribonucleic acid (mRNA) of IL-17, IL-22, and IL-1β. The effects of CT on H₂O₂-induced HTR-8/SVneo cells were investigated using cell counting kit-8 (CCK-8) assays and cell migration assays, while flow cytometry was used for the detection of apoptosis. Network pharmacology was used to analyze the mechanism of CT in the treatment of URSA. The repair effects and underlying mechanisms of the tumor protein 53 (p53) inhibitor and signal and transducer of transcription 3 small interfering RNA (STAT3 siRNA) were evaluated using ELISA assays, CCK-8, cell migration assays, flow cytometry, immunofluorescence, and Western blot techniques.

Results

Different doses of CT exerted protective effects on the uterine tissues of URSA. Also, CT could reduce levels of IL-17, IL-22, and IL-1β and their relative mRNA content in the serum of URSA mice, as well as increase the activity and migration rate of HTR-8/SVneo cells, and reduce the rate of apoptosis. Using p53 and STAT3 as mechanistic targets, we found that the p53 inhibitor, STAT3 siRNA, and CT restored the migratory activity of HTR-8/SVneo cells, reduced their apoptotic rate, and simultaneously downregulated the expression of retinoic acid receptor-related orphan receptor γt (RORγt) and vascular endothelial growth factor (VEGF).

Conclusion

The mechanism by which CT regulated the activity of HTR-8/SVneo cells involved the inhibition of p-p53, with STAT3, RORγt, and VEGF functioning as downstream targets of the p53 pathway.

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隐丹参酮通过p53/STAT3通路调控HTR-8/SVneo细胞活性在不明原因复发性自然流产中的作用

目的：探讨隐丹参酮（CT）治疗不明原因复发性自然流产（URSA）的疗效及机制。方法：在高、中、低剂量CT治疗前，将DBA/2雄性小鼠与CBA/J雌性小鼠笼合，建立URSA模型。采用酶联免疫吸附法（ELISA）检测各组小鼠血清中白细胞介素(IL)-1 β (β)、IL-17和IL-22的水平。采用苏木精-伊红（HE）病理检测蜕膜和子宫组织，采用聚合酶链反应（PCR）检测血清信使核糖核酸（mRNA）中IL-17、IL-22和IL-1β的相对表达水平。采用细胞计数试剂盒-8 （CCK-8）法和细胞迁移法观察CT对h2o2诱导HTR-8/SVneo细胞的影响，流式细胞术检测细胞凋亡情况。采用网络药理学方法分析CT治疗URSA的机制。使用ELISA、CCK-8、细胞迁移、流式细胞术、免疫荧光和Western blot技术评估肿瘤蛋白53 （p53）抑制剂和转录小干扰RNA （STAT3 siRNA）的修复作用和潜在机制。结果：不同剂量CT对URSA子宫组织均有保护作用。CT可降低URSA小鼠血清中IL-17、IL-22、IL-1β水平及其相对mRNA含量，提高HTR-8/SVneo细胞活性和迁移率，降低凋亡率。以p53和STAT3为机制靶点，我们发现p53抑制剂、STAT3 siRNA和CT可恢复HTR-8/SVneo细胞的迁移活性，降低其凋亡率，同时下调视黄酸受体相关孤儿受体γt （RORγt）和血管内皮生长因子（VEGF）的表达。结论：CT调节HTR-8/SVneo细胞活性的机制与抑制p-p53有关，STAT3、RORγt和VEGF是p53通路的下游靶点。

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来源期刊

American Journal of Reproductive Immunology 医学-免疫学

CiteScore

6.20

自引率

5.60%

发文量

314

审稿时长

2 months

期刊介绍： The American Journal of Reproductive Immunology is an international journal devoted to the presentation of current information in all areas relating to Reproductive Immunology. The journal is directed toward both the basic scientist and the clinician, covering the whole process of reproduction as affected by immunological processes. The journal covers a variety of subspecialty topics, including fertility immunology, pregnancy immunology, immunogenetics, mucosal immunology, immunocontraception, endometriosis, abortion, tumor immunology of the reproductive tract, autoantibodies, infectious disease of the reproductive tract, and technical news.