Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells
Abstract Background Hyaluronan is the main component of the cumulus-oocyte complex (COC) matrix, and it maintains the basic structure of the COC during ovulation. As a member of the transforming growth factor β (TGF-β) superfamily, bone morphogenetic protein 2 (BMP2) has been identified as a critica...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-04-01
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| Series: | Reproductive Biology and Endocrinology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12958-022-00937-y |
| _version_ | 1819022641127751680 |
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| author | Hsun-Ming Chang Long Bai Yi-Min Zhu Peter C. K. Leung |
| author_facet | Hsun-Ming Chang Long Bai Yi-Min Zhu Peter C. K. Leung |
| author_sort | Hsun-Ming Chang |
| collection | DOAJ |
| description | Abstract Background Hyaluronan is the main component of the cumulus-oocyte complex (COC) matrix, and it maintains the basic structure of the COC during ovulation. As a member of the transforming growth factor β (TGF-β) superfamily, bone morphogenetic protein 2 (BMP2) has been identified as a critical regulator of mammalian folliculogenesis and ovulation. However, whether BMP2 can regulate the production of hyaluronan in human granulosa cells has never been elucidated. Methods In the present study, we investigated the effect of BMP2 on the production of hyaluronan and the underlying molecular mechanism using both immortalized (SVOG) and primary human granulosa-lutein (hGL) cells. The expression of three hyaluronan synthases (including HAS1, HAS2 and HAS3) were examined following cell incubation with BMP2 at different concentrations. The concentrations of the hyaluronan cell culture medium were determined by enzyme-linked immunosorbent assay (ELISA). The TGF-β type I receptor inhibitors (dorsomorphin and DMH-1) and small interfering RNAs targeting ALK2, ALK3, ALK6 and SMAD4 were used to investigate the involvement of TGF-β type I receptor and SMAD-dependent pathway. Results Our results showed that BMP2 treatment significantly increased the production of hyaluronan by upregulating the expression of hyaluronan synthase 2 (HAS2). In addition, BMP2 upregulates the expression of connective tissue growth factor (CTGF), which subsequently mediates the BMP2-induced increases in HAS2 expression and hyaluronan production because overexpression of CTGF enhances, whereas knockdown of CTGF reverses, these effects. Notably, using kinase inhibitor- and siRNA-mediated knockdown approaches, we demonstrated that the inductive effect of BMP2 on the upregulation of CTGF is mediated by the ALK2/ALK3-mediated SMAD-dependent signaling pathway. Conclusions Our findings provide new insight into the molecular mechanism by which BMP2 promotes the production of hyaluronan in human granulosa cells. |
| first_indexed | 2024-12-21T04:26:13Z |
| format | Article |
| id | doaj.art-0ea123a3e67c4c8abed6e453b1a7a60d |
| institution | Directory Open Access Journal |
| issn | 1477-7827 |
| language | English |
| last_indexed | 2024-12-21T04:26:13Z |
| publishDate | 2022-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Reproductive Biology and Endocrinology |
| spelling | doaj.art-0ea123a3e67c4c8abed6e453b1a7a60d2022-12-21T19:16:03ZengBMCReproductive Biology and Endocrinology1477-78272022-04-0120111310.1186/s12958-022-00937-yConnective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cellsHsun-Ming Chang0Long Bai1Yi-Min Zhu2Peter C. K. Leung3Department of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang UniversityDepartment of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang UniversityDepartment of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang UniversityDepartment of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang UniversityAbstract Background Hyaluronan is the main component of the cumulus-oocyte complex (COC) matrix, and it maintains the basic structure of the COC during ovulation. As a member of the transforming growth factor β (TGF-β) superfamily, bone morphogenetic protein 2 (BMP2) has been identified as a critical regulator of mammalian folliculogenesis and ovulation. However, whether BMP2 can regulate the production of hyaluronan in human granulosa cells has never been elucidated. Methods In the present study, we investigated the effect of BMP2 on the production of hyaluronan and the underlying molecular mechanism using both immortalized (SVOG) and primary human granulosa-lutein (hGL) cells. The expression of three hyaluronan synthases (including HAS1, HAS2 and HAS3) were examined following cell incubation with BMP2 at different concentrations. The concentrations of the hyaluronan cell culture medium were determined by enzyme-linked immunosorbent assay (ELISA). The TGF-β type I receptor inhibitors (dorsomorphin and DMH-1) and small interfering RNAs targeting ALK2, ALK3, ALK6 and SMAD4 were used to investigate the involvement of TGF-β type I receptor and SMAD-dependent pathway. Results Our results showed that BMP2 treatment significantly increased the production of hyaluronan by upregulating the expression of hyaluronan synthase 2 (HAS2). In addition, BMP2 upregulates the expression of connective tissue growth factor (CTGF), which subsequently mediates the BMP2-induced increases in HAS2 expression and hyaluronan production because overexpression of CTGF enhances, whereas knockdown of CTGF reverses, these effects. Notably, using kinase inhibitor- and siRNA-mediated knockdown approaches, we demonstrated that the inductive effect of BMP2 on the upregulation of CTGF is mediated by the ALK2/ALK3-mediated SMAD-dependent signaling pathway. Conclusions Our findings provide new insight into the molecular mechanism by which BMP2 promotes the production of hyaluronan in human granulosa cells.https://doi.org/10.1186/s12958-022-00937-yBMP2CTGFHAS2HyaluronanHuman granulosa cells |
| spellingShingle | Hsun-Ming Chang Long Bai Yi-Min Zhu Peter C. K. Leung Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells Reproductive Biology and Endocrinology BMP2 CTGF HAS2 Hyaluronan Human granulosa cells |
| title | Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells |
| title_full | Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells |
| title_fullStr | Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells |
| title_full_unstemmed | Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells |
| title_short | Connective tissue growth factor mediates bone morphogenetic protein 2-induced increase in hyaluronan production in luteinized human granulosa cells |
| title_sort | connective tissue growth factor mediates bone morphogenetic protein 2 induced increase in hyaluronan production in luteinized human granulosa cells |
| topic | BMP2 CTGF HAS2 Hyaluronan Human granulosa cells |
| url | https://doi.org/10.1186/s12958-022-00937-y |
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