Wuzi-Yanzong-Wan Prevents the Defect of Cell-Cell Junctions between Sertoli-Germ Cells by Up-Regulating the Expression of TAp73.

IF 1.6 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS
Li Li, Min Pan, Ziao Liu, Hongsu Zhao, Fengqing Xu, Xiaohui Tong, Yajuan Wang, Bin Chen, Lihua Yu, Tongsheng Wang
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引用次数: 0

Abstract

Background: The TAp73 gene is an anti-cancer gene that also affects the junction between Sertoli and germ cells. Inhibition of this gene causes infertility in male mice. Our previous research proved that Wuzi-Yanzong-Wan (WZYZW) can protect spermatogenesis and maturation by preventing TAp73 inhibition.

Objective: This study aimed to investigate the effect of drug-containing serum of WZYZW on the defect of cell-cell junctions in the Sertoli-germ cells co-culture system in vitro.

Methods: LC-HRMS was used to analyze the content of active ingredients in WZYZWmedicated serum. Then, primary extraction and co-culture of germ cells and Sertoli cells were carried out. Co-cultured cells were added with PFT-α to induce the TAp73 inhibition model, with WZYZW-medicated serum at 2.5%, 5%, and 10% treated in parallel. Sloughing of germ cells from Sertoli cells was calculated. Transmission electron microscopy (TEM), Immunofluorescence, qRT-PCR, and western blot methods were employed.

Results: The drug-containing serum of WZYZW contained schisandrin, hyperoside, geniposidic acid, ellagic acid, and quercetin. Using TEM assay, we observed restoration of the desmosomelike (Des), tight junctions (TJ), and basal ectoplasmic specialization (ES) structure following WZYZW treatment. WZYZW caused inhibition of peptidase and protease inhibitors (tissue inhibitor of metalloproteinase-1 (TIMP1), Serpina3n) by immunofluorescence analysis. Western blot and qRT-PCR analysis revealed that WZYZW was able to ameliorate the expressions of peptidase and protease inhibitors and cell adhesion factors, such as TAp73, TIMP1, Serpina3n, Desmocollin-3, N-cadherin, and Nectin-2.

Conclusion: WZYZW-medicated serum could prevent the defect of cell-cell junctions between Sertoli-germ cells co-culture system in vitro by up-regulating the expression of TAp73.

五子衍宗丸通过上调 TAp73 的表达防止 Sertoli-Germ 细胞间的细胞-细胞连接缺陷
背景:TAp73 基因是一种抗癌基因,也会影响 Sertoli 细胞和生殖细胞之间的连接。抑制该基因会导致雄性小鼠不育。我们之前的研究证明,五子衍宗丸(WZYZW)可通过阻止 TAp73 的抑制作用来保护精子发生和成熟:本研究旨在探讨五子衍宗丸含药血清对体外Sertoli-germ细胞共培养系统中细胞-细胞连接缺陷的影响:方法:采用LC-HRMS分析WZYZW药用血清中有效成分的含量。然后,对生殖细胞和 Sertoli 细胞进行原代提取和共培养。在共培养细胞中加入 PFT-α,诱导 TAp73 抑制模型,并同时处理 2.5%、5% 和 10%的 WZYZW 给药血清。计算Sertoli细胞中生殖细胞的脱落情况。研究采用了透射电子显微镜(TEM)、免疫荧光、qRT-PCR和Western印迹等方法:结果:WZYZW的含药血清中含有五味子素、金丝桃苷、玄参苷酸、鞣花酸和槲皮素。通过TEM检测,我们观察到WZYZW处理后的脱丝体(Des)、紧密连接(TJ)和基底外质特化(ES)结构得到恢复。通过免疫荧光分析,WZYZW 可抑制肽酶和蛋白酶抑制剂(金属蛋白酶组织抑制剂-1(TIMP1)、Serpina3n)。Western印迹和qRT-PCR分析显示,WZYZW能够改善肽酶和蛋白酶抑制剂以及TAp73、TIMP1、Serpina3n、Desmocollin-3、N-cadherin和Nectin-2等细胞粘附因子的表达:结论:WZYZW药用血清可通过上调TAp73的表达,防止体外Sertol-germ细胞共培养系统细胞间连接的缺陷。
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来源期刊
CiteScore
3.10
自引率
5.60%
发文量
327
审稿时长
7.5 months
期刊介绍: Combinatorial Chemistry & High Throughput Screening (CCHTS) publishes full length original research articles and reviews/mini-reviews dealing with various topics related to chemical biology (High Throughput Screening, Combinatorial Chemistry, Chemoinformatics, Laboratory Automation and Compound management) in advancing drug discovery research. Original research articles and reviews in the following areas are of special interest to the readers of this journal: Target identification and validation Assay design, development, miniaturization and comparison High throughput/high content/in silico screening and associated technologies Label-free detection technologies and applications Stem cell technologies Biomarkers ADMET/PK/PD methodologies and screening Probe discovery and development, hit to lead optimization Combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) Chemical library design and chemical diversity Chemo/bio-informatics, data mining Compound management Pharmacognosy Natural Products Research (Chemistry, Biology and Pharmacology of Natural Products) Natural Product Analytical Studies Bipharmaceutical studies of Natural products Drug repurposing Data management and statistical analysis Laboratory automation, robotics, microfluidics, signal detection technologies Current & Future Institutional Research Profile Technology transfer, legal and licensing issues Patents.
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