益肾通络方的 microRNAs 可以修复苯并芘引起的精子 DNA 损伤。

MicroRNAs from Yishen Tongluo formula can repair sperm DNA damage caused by benzo()pyrene.

机构信息

The Second Clinical Medical School, Henan University of Chinese Medicine, Zhengzhou, Henan, China.

The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, Henan, China.

出版信息

Pharm Biol. 2024 Dec;62(1):781-789. doi: 10.1080/13880209.2024.2417002. Epub 2024 Oct 22.

DOI:10.1080/13880209.2024.2417002

PMID:39435988

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11497566/

Abstract

CONTEXT

Plant microRNAs (miRNAs) present in Yishen Tongluo formula (YSTL, a traditional Chinese herbal medicine formula) are considered as potential therapeutic drugs for reducing the sperm DNA fragmentation index (DFI).

OBJECTIVE

To study the effectiveness of plant miRNAs in YSTL for repairing mouse sperm DNA damage caused by benzo()pyrene (BaP).

METHODS AND MATERIALS

Twenty-four male SPF ICR (CD1) mice were divided into control, BaP and YSTL groups. A BaP-induced (100 mg/kg) sperm DNA damage model was established in the BaP and YSTL groups, and the mice in the YSTL group were treated with YSTL (23.78 g/kg) for 8 weeks. Sperm DFI was determined a sperm chromatin structure assay (SCSA). MicroRNAs in the testes of the mice were analysed RNA-seq, and the top four plant miRNAs were screened, identified and overexpressed in GC cells. The effects of plant miRNAs on the viability and DNA integrity of GC cells exposed to benzo()pyrene diol epoxide (BPDE) (1 μM) were tested using CCK8 and comet assays.

RESULTS

Compared with that of the BaP group, the DFI of the YSTL group decreased (9.57% vs. 18.54%, = 18.645, = 0.0236). miR166-y, miR894-x, miR822-x and miR396-x were screened. The CCK8 and comet assays revealed that the DFI of the mimic group was significantly lower than that of the BPDE (IC = 1.006 μM) group, with the most significant difference in the miR396-x group.

DISCUSSION AND CONCLUSIONS

Plant miRNAs such as miR396-x can penetrate the blood-testis barrier through the digestive system to repair sperm DNA.

摘要

背景

益肾通络方（YSTL，一种中药方剂）中的植物 microRNAs（miRNAs）被认为是降低精子 DNA 碎片化指数（DFI）的潜在治疗药物。

目的

研究 YSTL 中的植物 miRNAs 修复苯并（a）芘（BaP）引起的小鼠精子 DNA 损伤的效果。

方法和材料

将 24 只雄性 SPF ICR（CD1）小鼠分为对照组、BaP 组和 YSTL 组。在 BaP 组和 YSTL 组中建立 BaP 诱导的（100mg/kg）精子 DNA 损伤模型，YSTL 组用 YSTL（23.78g/kg）治疗 8 周。通过精子染色质结构分析（SCSA）测定精子 DFI。通过 RNA-seq 分析小鼠睾丸中的 miRNAs，并筛选、鉴定和过表达前 4 种植物 miRNAs 在 GC 细胞中。使用 CCK8 和彗星试验测试植物 miRNAs 对暴露于苯并（a）芘二醇环氧化物（BPDE）（1μM）的 GC 细胞活力和 DNA 完整性的影响。

结果

与 BaP 组相比，YSTL 组的 DFI 降低（9.57%比 18.54%，=18.645，=0.0236）。筛选出 miR166-y、miR894-x、miR822-x 和 miR396-x。CCK8 和彗星试验显示，miR396-x 组的 mimic 组的 DFI 明显低于 BPDE（IC=1.006μM）组，差异最显著。