Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative.
Adipocytes attached to the extracellular matrix (ECM) mainly consist of collagen in adipose tissues, while the degradation of ECM by collagenase induces the apoptosis of adipocytes, leading to a decrease in local subcutaneous adipose. To achieve this goal, we are developing a mutant collagenase H (C...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2019-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227202&type=printable |
| _version_ | 1826574238672224256 |
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| author | Fuding Chen Gang Du Meishu Shih Hongjiang Yuan Peng Bao Sheng Shi Yong Cang Zhen Zhang |
| author_facet | Fuding Chen Gang Du Meishu Shih Hongjiang Yuan Peng Bao Sheng Shi Yong Cang Zhen Zhang |
| author_sort | Fuding Chen |
| collection | DOAJ |
| description | Adipocytes attached to the extracellular matrix (ECM) mainly consist of collagen in adipose tissues, while the degradation of ECM by collagenase induces the apoptosis of adipocytes, leading to a decrease in local subcutaneous adipose. To achieve this goal, we are developing a mutant collagenase H (ColH) to remove local subcutaneous fat such as submental fat (SMF). Three vectors were constructed for expressing rColH(FM, mutant for fat melting, with 6xHis tag), rColH(WT, wild-type, with 6xHis tag), and rColH(E451D, E451D mutant, without 6xHis tag) in Escherichia coli. rColH(FM) & rColH(WT) were purified by Ni Sepharose on a laboratory scale, while rColH(E451D) was purified by five chromatography purification steps on a large scale. Then, the stability of rColH(FM) and rColH(WT) was tested by SDS-PAGE to investigate the influence of the E451D mutation on stability. Afterwards, the enzyme kinetics of ColH (mutant or wild-type, with or without His tag) were investigated and compared. Finally, the adipolysis of rColH(E451D) at various doses was tested in vitro and in vivo. The ultrasound results in minipigs suggested that effective adipolysis was induced by rColH(E451D) compared with the negative control, and the histological results suggest dose-dependent fibrosis, necrosis, inflammation and cholesterol cleft formation. These findings indicate the possibility of rColH(E451D) becoming a new injectable drug to safely remove subcutaneous adipose. |
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| id | doaj.art-d4fb2e3fe46a474988a53d3bab776683 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2025-03-14T12:55:38Z |
| publishDate | 2019-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d4fb2e3fe46a474988a53d3bab7766832025-03-02T05:33:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-011412e022720210.1371/journal.pone.0227202Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative.Fuding ChenGang DuMeishu ShihHongjiang YuanPeng BaoSheng ShiYong CangZhen ZhangAdipocytes attached to the extracellular matrix (ECM) mainly consist of collagen in adipose tissues, while the degradation of ECM by collagenase induces the apoptosis of adipocytes, leading to a decrease in local subcutaneous adipose. To achieve this goal, we are developing a mutant collagenase H (ColH) to remove local subcutaneous fat such as submental fat (SMF). Three vectors were constructed for expressing rColH(FM, mutant for fat melting, with 6xHis tag), rColH(WT, wild-type, with 6xHis tag), and rColH(E451D, E451D mutant, without 6xHis tag) in Escherichia coli. rColH(FM) & rColH(WT) were purified by Ni Sepharose on a laboratory scale, while rColH(E451D) was purified by five chromatography purification steps on a large scale. Then, the stability of rColH(FM) and rColH(WT) was tested by SDS-PAGE to investigate the influence of the E451D mutation on stability. Afterwards, the enzyme kinetics of ColH (mutant or wild-type, with or without His tag) were investigated and compared. Finally, the adipolysis of rColH(E451D) at various doses was tested in vitro and in vivo. The ultrasound results in minipigs suggested that effective adipolysis was induced by rColH(E451D) compared with the negative control, and the histological results suggest dose-dependent fibrosis, necrosis, inflammation and cholesterol cleft formation. These findings indicate the possibility of rColH(E451D) becoming a new injectable drug to safely remove subcutaneous adipose.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227202&type=printable |
| spellingShingle | Fuding Chen Gang Du Meishu Shih Hongjiang Yuan Peng Bao Sheng Shi Yong Cang Zhen Zhang Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. PLoS ONE |
| title | Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. |
| title_full | Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. |
| title_fullStr | Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. |
| title_full_unstemmed | Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. |
| title_short | Safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative. |
| title_sort | safe and effective subcutaneous adipolysis in minipigs by a collagenase derivative |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227202&type=printable |
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