Zhejiang Vocational College of Special Education, Hangzhou, China.
Women's Hospital School of Medicine, Zhejiang University, Hangzhou, China.
Pharm Biol. 2023 Dec;61(1):1387-1400. doi: 10.1080/13880209.2023.2249193.
di-(2-Ethylhexyl) phthalate (DEHP) has potential reproductive toxicity. Bu-Shen-Tian-Jing formulations (BSTJFs) are beneficial for female reproductive capacity. However, BSTJF2 has much lower cytotoxicity than BSTJF1.
To investigate the effects of BSTJFs on ovarian granulosa cells exposed to DEHP and determine the potential molecular mechanisms.
Human granulosa-like tumor cell line (KGN) cells were divided into control, DEHP, BSTJF1 and BSTJF2 groups. The DEHP group were given 1 μM DEHP for 24 h. They were then given BSTJF1 at 200 μg/mL or BSTJF2 at 100 μg/mL for 24 h. The control group was treated with the same concentration of DMSO (0.1%). Oxidative stress and mitochondrial function were measured. The mRNA and protein expression levels of HDAC3 and HSP90AA were determined. Integrative network pharmacology analysis of BSTJF2 was also performed.
DEHP (1 μM) significantly suppressed the proliferation of KGN cells by 17%, significantly increased ROS levels by 28% and MDA levels by 47%, significantly decreased MMP levels by 22% and mtDNA copy by 30%. DEHP significantly increased protein expression of HDAC3 by 21%and HSP90AA by 64%. All these changes were significantly reversed by BSTJFs. Integrative network pharmacology analysis revealed HSP90AA was a key target (degree = 8). Both RGFP966 and BSTJF2 significantly reversed the increased expression of HDAC3 and HSP90AA, attenuated oxidative stress, and mitochondrial damage which were induced by DEHP.
BSTJFs might have therapeutic potential on oxidative stress and mitochondrial damage through the HDAC3/HSP90AA pathway which encourages further clinical trials.
邻苯二甲酸二（2-乙基己基）酯（DEHP）具有潜在的生殖毒性。补肾填精方（BSTJF）有益于女性生殖能力。然而，BSTJF2 的细胞毒性比 BSTJF1 低得多。
研究 BSTJF 对 DEHP 暴露的卵巢颗粒细胞的影响，并确定其潜在的分子机制。
将人卵巢颗粒样瘤细胞系（KGN）细胞分为对照组、DEHP 组、BSTJF1 组和 BSTJF2 组。DEHP 组给予 1μM DEHP 处理 24h，然后分别给予 BSTJF1 200μg/ml 或 BSTJF2 100μg/ml 处理 24h。对照组用相同浓度的 DMSO（0.1%）处理。检测氧化应激和线粒体功能，测定 HDAC3 和 HSP90AA 的 mRNA 和蛋白表达水平。对 BSTJF2 进行整合网络药理学分析。
1μM 的 DEHP 使 KGN 细胞的增殖率显著降低了 17%，ROS 水平显著升高了 28%，MDA 水平显著升高了 47%，MMP 水平显著降低了 22%，mtDNA 拷贝数显著降低了 30%。DEHP 使 HDAC3 蛋白表达显著增加了 21%，HSP90AA 蛋白表达显著增加了 64%。BSTJF2 可显著逆转上述变化。整合网络药理学分析表明 HSP90AA 是一个关键靶点（度=8）。RGFP966 和 BSTJF2 均可显著逆转 DEHP 诱导的 HDAC3 和 HSP90AA 表达增加、氧化应激和线粒体损伤。
BSTJF 可能通过 HDAC3/HSP90AA 通路对氧化应激和线粒体损伤具有治疗潜力，值得进一步开展临床试验。
