Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts
AIM: To study the effect of zymosan, a ligand found on the surface of fungi, on gap junctional intercellular communication (GJIC) in cultured human corneal fibroblasts (HCFs). METHODS: Zymosan was added to the medium of cultured HCFs with or without the administration of mitogen-activated protein ki...
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| Format: | Article |
| Language: | English |
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Press of International Journal of Ophthalmology (IJO PRESS)
2021-03-01
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| Series: | International Journal of Ophthalmology |
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| Online Access: | http://ies.ijo.cn/en_publish/2021/3/20210302.pdf |
| _version_ | 1831641102493417472 |
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| author | Xiao-Shuo Zheng Hui Zheng Dan Xu Ping-Ping Liu Bing Li Zi-Mu Cao Yang Liu Ye Liu |
| author_facet | Xiao-Shuo Zheng Hui Zheng Dan Xu Ping-Ping Liu Bing Li Zi-Mu Cao Yang Liu Ye Liu |
| author_sort | Xiao-Shuo Zheng |
| collection | DOAJ |
| description | AIM: To study the effect of zymosan, a ligand found on the surface of fungi, on gap junctional intercellular communication (GJIC) in cultured human corneal fibroblasts (HCFs). METHODS: Zymosan was added to the medium of cultured HCFs with or without the administration of mitogen-activated protein kinase (MAPK) inhibitors or the inhibitor kappa B kinase 2 (IKK2) inhibitor IV. The protein and mRNA levels of connexin 43 (Cx43) in HCFs were measured by Western blot, immunofluorescence, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses. The GJIC activity was tested using a dye-coupling assay. RESULTS: The reduction of Cx43 protein and mRNA levels as well as a significant decrease in GJIC activity were observed in cultured HCFs when zymosan was added into the culture medium. Compared with controls (no zymosan), the protein level of Cx43 was reduced by 45% and 54% in the presence of zymosan at 200 and 600 μg/mL, respectively (P<0.05); and it was reduced by 45%, 48%, and 75% in the presence of zymosan (600 μg/mL) for 24, 36, and 48h, respectively (P<0.05). The mRNA expression of Cx43 was reduced by 98% in the presence of zymosan (P<0.05). The effects of zymosan on Cx43 expression and GJIC activity were attenuated by the administration of PD98059 [an extracellular signal-regulated kinase (ERK) signaling inhibitor] (P<0.05), c-Jun NH2-terminal kinase (JNK) inhibitor II (P<0.05), and IKK2 inhibitor IV (P<0.05). CONCLUSION: Zymosan inhibits the activity of GJIC in cultured HCFs. This effect is likely regulated via the nuclear factor-κB (NF-κB), MAPK/ERK, and JNK signaling pathways. The inhibitory effects of zymosan on Cx43 expression and GJIC activity in HCFs may induce damage of corneal stroma during corneal fungal infection. |
| first_indexed | 2024-12-19T12:19:27Z |
| format | Article |
| id | doaj.art-0c3338539ea64592840f71fa4006d6ee |
| institution | Directory Open Access Journal |
| issn | 2222-3959 2227-4898 |
| language | English |
| last_indexed | 2024-12-19T12:19:27Z |
| publishDate | 2021-03-01 |
| publisher | Press of International Journal of Ophthalmology (IJO PRESS) |
| record_format | Article |
| series | International Journal of Ophthalmology |
| spelling | doaj.art-0c3338539ea64592840f71fa4006d6ee2022-12-21T20:21:51ZengPress of International Journal of Ophthalmology (IJO PRESS)International Journal of Ophthalmology2222-39592227-48982021-03-0114334134810.18240/ijo.2021.03.0220210302Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblastsXiao-Shuo Zheng0Hui Zheng1Dan Xu2Ping-Ping Liu3Bing Li4Zi-Mu Cao5Yang Liu6Ye Liu7Ye Liu. Department of Pathology, the Fifth Affiliated Hospital of Sun Yat-sen University, 52 Road Meihuadong, Zhuhai 519000, Guangzhou Province, China. liuye23@mail.sysu.edu.cnDepartment of Ophthalmology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangzhou Province, ChinaInstitute of Environmental Systems Biology, Environmental Science and Engineering College, Dalian Maritime University, Dalian 116027, Liaoning Province, ChinaDepartment of Ophthalmology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangzhou Province, ChinaDepartment of Ophthalmology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangzhou Province, ChinaInstitute of Environmental Systems Biology, Environmental Science and Engineering College, Dalian Maritime University, Dalian 116027, Liaoning Province, ChinaDepartment of Ophthalmology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangzhou Province, ChinaDepartment of Pathology, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, Guangzhou Province, ChinaAIM: To study the effect of zymosan, a ligand found on the surface of fungi, on gap junctional intercellular communication (GJIC) in cultured human corneal fibroblasts (HCFs). METHODS: Zymosan was added to the medium of cultured HCFs with or without the administration of mitogen-activated protein kinase (MAPK) inhibitors or the inhibitor kappa B kinase 2 (IKK2) inhibitor IV. The protein and mRNA levels of connexin 43 (Cx43) in HCFs were measured by Western blot, immunofluorescence, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses. The GJIC activity was tested using a dye-coupling assay. RESULTS: The reduction of Cx43 protein and mRNA levels as well as a significant decrease in GJIC activity were observed in cultured HCFs when zymosan was added into the culture medium. Compared with controls (no zymosan), the protein level of Cx43 was reduced by 45% and 54% in the presence of zymosan at 200 and 600 μg/mL, respectively (P<0.05); and it was reduced by 45%, 48%, and 75% in the presence of zymosan (600 μg/mL) for 24, 36, and 48h, respectively (P<0.05). The mRNA expression of Cx43 was reduced by 98% in the presence of zymosan (P<0.05). The effects of zymosan on Cx43 expression and GJIC activity were attenuated by the administration of PD98059 [an extracellular signal-regulated kinase (ERK) signaling inhibitor] (P<0.05), c-Jun NH2-terminal kinase (JNK) inhibitor II (P<0.05), and IKK2 inhibitor IV (P<0.05). CONCLUSION: Zymosan inhibits the activity of GJIC in cultured HCFs. This effect is likely regulated via the nuclear factor-κB (NF-κB), MAPK/ERK, and JNK signaling pathways. The inhibitory effects of zymosan on Cx43 expression and GJIC activity in HCFs may induce damage of corneal stroma during corneal fungal infection.http://ies.ijo.cn/en_publish/2021/3/20210302.pdfcorneal fibroblastzymosangap junctionconnexin 43mapks |
| spellingShingle | Xiao-Shuo Zheng Hui Zheng Dan Xu Ping-Ping Liu Bing Li Zi-Mu Cao Yang Liu Ye Liu Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts International Journal of Ophthalmology corneal fibroblast zymosan gap junction connexin 43 mapks |
| title | Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts |
| title_full | Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts |
| title_fullStr | Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts |
| title_full_unstemmed | Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts |
| title_short | Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts |
| title_sort | effect of zymosan on the expression and function of the gap junction protein connexin 43 in human corneal fibroblasts |
| topic | corneal fibroblast zymosan gap junction connexin 43 mapks |
| url | http://ies.ijo.cn/en_publish/2021/3/20210302.pdf |
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