Ginsenoside Rg3 protects mouse leydig cells against triptolide by downregulation of miR-26a
Identifieur interne : 000801 ( Ncbi/Curation ); précédent : 000800; suivant : 000802Ginsenoside Rg3 protects mouse leydig cells against triptolide by downregulation of miR-26a
Auteurs : Haiyan Liang [République populaire de Chine] ; Suwei Zhang [République populaire de Chine] ; Zhiling Li [République populaire de Chine]Source :
- Drug Design, Development and Therapy [ 1177-8881 ] ; 2019.
Abstract
Ginsenoside Rg3 has been reported to exert protection function on germ cells. However, the mechanisms by which Rg3 regulates apoptosis in mouse Leydig cells remain unclear. In addition, triptolide (TP) has been reported to induce infertility in male rats. Thus, this study aimed to investigate the protective effect of Rg3 against TP-induced toxicity in MLTC-1 cells.
CCK-8, immunofluorescence assay, Western blotting and flow cytometry were used to detect cell proliferation and cell apoptosis, respectively. In addition, the dual luciferase reporter system assay was used to detect the interaction between miR-26a and GSK3β in MLTC-1 cells.
TP significantly inhibited the proliferation of MLTC-1 cells, while the inhibitory effect of TP was reversed by Rg3. In addition, TP markedly induced apoptosis in MLTC-1 cells via increasing the expressions of Bax, active caspase 3, Cyto c and active caspase 9, and decreasing the level of Bcl-2. However, Rg3 alleviated TP-induced apoptosis of MLTC-1 cells. Moreover, the level of miR-26a was obviously downregulated by Rg3 treatment. The protective effect of Rg3 against TP-induced toxicity in MLTC-1 cells was abolished by miR-26a upregulation. Meanwhile, dual-luciferase assay showed GSK3β was the direct target of miR-26a in MLTC-1 cells. Overexpression of miR-26a markedly decreased the level of GSK3β. As expected, upregulation of miR-26a could abrogate the protective effects of Rg3 against TP-induced cytotoxicity via inhibiting the expression of GSK3β.
These results indicated that Rg3 could protect MLTC-1 against TP by downregulation of miR-26a. Therefore, Rg3 might serve as a potential agent for the treatment of male hypogonadism.
Url:
DOI: 10.2147/DDDT.S208328
PubMed: 31296984
PubMed Central: 6598939
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<author><name sortKey="Liang, Haiyan" sort="Liang, Haiyan" uniqKey="Liang H" first="Haiyan" last="Liang">Haiyan Liang</name>
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,<country>People’s Republic of China</country>
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<country xml:lang="fr">République populaire de Chine</country>
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<author><name sortKey="Zhang, Suwei" sort="Zhang, Suwei" uniqKey="Zhang S" first="Suwei" last="Zhang">Suwei Zhang</name>
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<author><name sortKey="Li, Zhiling" sort="Li, Zhiling" uniqKey="Li Z" first="Zhiling" last="Li">Zhiling Li</name>
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<author><name sortKey="Zhang, Suwei" sort="Zhang, Suwei" uniqKey="Zhang S" first="Suwei" last="Zhang">Suwei Zhang</name>
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<front><div type="abstract" xml:lang="en"><sec id="S2001"><title>Background</title>
<p>Ginsenoside Rg3 has been reported to exert protection function on germ cells. However, the mechanisms by which Rg3 regulates apoptosis in mouse Leydig cells remain unclear. In addition, triptolide (TP) has been reported to induce infertility in male rats. Thus, this study aimed to investigate the protective effect of Rg3 against TP-induced toxicity in MLTC-1 cells.</p>
</sec>
<sec id="S2002"><title>Methods</title>
<p>CCK-8, immunofluorescence assay, Western blotting and flow cytometry were used to detect cell proliferation and cell apoptosis, respectively. In addition, the dual luciferase reporter system assay was used to detect the interaction between miR-26a and GSK3β in MLTC-1 cells.</p>
</sec>
<sec id="S2003"><title>Results</title>
<p>TP significantly inhibited the proliferation of MLTC-1 cells, while the inhibitory effect of TP was reversed by Rg3. In addition, TP markedly induced apoptosis in MLTC-1 cells via increasing the expressions of Bax, active caspase 3, Cyto c and active caspase 9, and decreasing the level of Bcl-2. However, Rg3 alleviated TP-induced apoptosis of MLTC-1 cells. Moreover, the level of miR-26a was obviously downregulated by Rg3 treatment. The protective effect of Rg3 against TP-induced toxicity in MLTC-1 cells was abolished by miR-26a upregulation. Meanwhile, dual-luciferase assay showed GSK3β was the direct target of miR-26a in MLTC-1 cells. Overexpression of miR-26a markedly decreased the level of GSK3β. As expected, upregulation of miR-26a could abrogate the protective effects of Rg3 against TP-induced cytotoxicity via inhibiting the expression of GSK3β.</p>
</sec>
<sec id="S2004"><title>Conclusion</title>
<p>These results indicated that Rg3 could protect MLTC-1 against TP by downregulation of miR-26a. Therefore, Rg3 might serve as a potential agent for the treatment of male hypogonadism.</p>
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