何首乌饮通过DNA甲基转移酶1延缓大鼠睾丸间质细胞衰老

吴恬; 路硕娅; 杨玉娇; 段豫磊; 安啟超; 甄晓兰; 牛嗣云

doi:10.16098/j.issn.0529-1356.2024.03.004

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解剖学报 ›› 2024, Vol. 55 ›› Issue (3) : 276-284. DOI: 10.16098/j.issn.0529-1356.2024.03.004

细胞和分子生物学

何首乌饮通过DNA甲基转移酶1延缓大鼠睾丸间质细胞衰老

吴恬¹ 路硕娅¹ 杨玉娇¹ 段豫磊¹ 安啟超¹ 甄晓兰^2* 牛嗣云^1*

作者信息 +

Heshouwuyin delaying the aging of Leydig cells in rat testis through DNA methyltransferase 1

WU Tian¹ LU Shuo-ya¹ YANG Yu-jiao¹ DUAN Yu-lei¹ AN Qi-chao¹ ZHEN Xiao-lan^2* NIU Si-yun^1*

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文章历史 +

摘要

目的探讨何首乌饮能否通过DNA甲基转移酶1（DNMT1）延缓大鼠睾丸间质细胞衰老。方法 40只Wistar雄性大鼠随机分为4组，每组10只。免疫组织化学检测各组大鼠睾丸组织中DNMT1表达水平。ELISA实验检测各组大鼠血清中的睾酮含量。通过自由基氧化损伤建立大鼠睾丸间质细胞衰老模型。在睾丸间质细胞中使用慢病毒敲低DNMT1，通过β-半乳糖苷酶(β-GAL)染色、免疫荧光染色和ELISA实验检测细胞衰老状态和细胞上清液中睾酮和睾酮合成关键酶3β羟基类固醇脱氢酶（3β-HSD）、细胞色素P450家族成员11A1（CYP11A1）的含量。结果与青年对照组相比，自然衰老组大鼠睾丸组织中P16蛋白表达和β-GAL阳性率明显升高，DNMT1表达和血清睾酮含量降低（P<0.05）；何首乌饮干预能够降低衰老大鼠睾丸组织P16蛋白表达和β-GAL阳性率，提高DNMT1表达和血清中的睾酮含量（P<0.05）。衰老的睾丸间质细胞中呈现相同的趋势。在睾丸间质细胞中敲低DNMT1后，β-GAL阳性率和P16蛋白表达明显增加，睾丸间质细胞的睾酮分泌量和睾酮合成关键酶3β-HSD、CYP11A1含量明显减少，与正常组相比差异具有统计学意义（P<0.05）。加入何首乌饮后，上述现象得到改善。结论何首乌饮可以通过DNMT1延缓大鼠睾丸间质细胞衰老。

Abstract

Objective To investigate whether Heshouwuyin can delay the aging of Leydig cells in rat testis through DNA methyltransferase 1 (DNMT1). Methods Totally 40 male Wistar rats were randomly divided into 4 groups, with 10 rats in each group. Immunohistochemistry was used to detect the expression levels of DNMT1 in testis tissue of rats. Testosterone content in serum of rats in each group was detected by ELISA test. A rat Leydig cell aging model was established by free radical oxidative damage. DNMT1 was knocked down by lentivirus in Leydig cells, and the cell senescence status and the testosterone content and testosterone synthesis key enzyme 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 family member 11A1 (CYP11A1) content secreted by cells were detected by β-GALactosidase（β-GAL）staining, immunofluorescenct staining and ELISA. Results Compared with the young control group (YCG), the expression of P16 protein and the positive rate of β-GAL in the testis tissue of rats in the natural aging group (NAG) increased significantly, and the expression of DNMT1 and serum testosterone levels decreased(P<0.05). However, after Heshouwuyin intervention, the expression of P16 protein and the positive rate of β-GAL in the testis of aging rats were reduced, and DNMT1 expression and the serum testosterone levels increased (P<0.05). The same trend was observed in Leydig cells. Knockdown of DNMT1 in Leydig cells, β-GAL positivity and P16 protein expression increased significantly, and testosterone secretion and testosterone synthesis key enzymes 3β-HSD, CYP11A1 content from Leydig cells decreased significantly, compared with the normal control group (NCG) (P<0.05). When Heshouwuyin was added, the above phenomenon was improved. Conclusion Heshouwuyin can delay the aging of rat Leydig cells through DNMT1.

导出引用

吴恬路硕娅杨玉娇段豫磊安啟超甄晓兰牛嗣云. 何首乌饮通过DNA甲基转移酶1延缓大鼠睾丸间质细胞衰老[J]. 解剖学报. 2024, 55(3): 276-284 https://doi.org/10.16098/j.issn.0529-1356.2024.03.004

WU Tian LU Shuo-ya YANG Yu-jiao DUAN Yu-lei AN Qi-chao ZHEN Xiao-lan NIU Si-yun. Heshouwuyin delaying the aging of Leydig cells in rat testis through DNA methyltransferase 1[J]. Acta Anatomica Sinica. 2024, 55(3): 276-284 https://doi.org/10.16098/j.issn.0529-1356.2024.03.004

中图分类号： Q291

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