Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis
Objective To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis. Methods The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of...
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| Veröffentlicht in: | Chinese journal of integrative medicine 2023-06, Vol.29 (6), p.508-516 |
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| creator | Ding, Dou Zhang, Qi Zeng, Fu-jia Cai, Ming-xing Gan, Yuan Dong, Xiao-jun |
| description | Objective
To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.
Methods
The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.
Results
The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (
P |
| doi_str_mv | 10.1007/s11655-022-3723-4 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2725441636</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2725441636</sourcerecordid><originalsourceid>FETCH-LOGICAL-c344t-9424c4f11806561a3c8bc8d13a56363099a10d03cf5a5d6b39e1cc1ff13362bb3</originalsourceid><addsrcrecordid>eNp9kM1OwzAQhC0EoqXwAFxQjlxMvf5LcwwVLUhFoArOluM41FWTFDuR4O1xCXDktLua2dHuh9AlkBsgJJ0GACkEJpRillKG-REaQ5YxTDihx7GXKY09iBE6C2FLiEglEadoxCQVABzG6PnRmo1uXKiTtkqWtulccCbJjSuTtknWG9vXumvjlPtu412Uk1sd7LdauzWGrMBsP13nC0jyDxfO0Umld8Fe_NQJel3cvczv8epp-TDPV9gwzjucccoNrwBmRAoJmplZYWYlMC0kk4xkmQZSEmYqoUUpC5ZZMAaqClg8vijYBF0PuXvfvvc2dKp2wdjdTje27YOiKRWcwyFsgmCwGt-G4G2l9t7V2n8qIOoAUg0gVQSpDiAVjztXP_F9Udvyb-OXXDTQwRCi1LxZr7Zt75v48j-pXxvLerw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2725441636</pqid></control><display><type>article</type><title>Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis</title><source>MEDLINE</source><source>Alma/SFX Local Collection</source><source>SpringerLink Journals - AutoHoldings</source><creator>Ding, Dou ; Zhang, Qi ; Zeng, Fu-jia ; Cai, Ming-xing ; Gan, Yuan ; Dong, Xiao-jun</creator><creatorcontrib>Ding, Dou ; Zhang, Qi ; Zeng, Fu-jia ; Cai, Ming-xing ; Gan, Yuan ; Dong, Xiao-jun</creatorcontrib><description>Objective
To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.
Methods
The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.
Results
The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (
P
<0.05 or
P
<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (
P
<0.05 or
P
<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (
P
<0.01).
Conclusion
GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.</description><identifier>ISSN: 1672-0415</identifier><identifier>EISSN: 1993-0402</identifier><identifier>DOI: 10.1007/s11655-022-3723-4</identifier><identifier>PMID: 36251141</identifier><language>eng</language><publisher>Singapore: Springer Nature Singapore</publisher><subject>Arthritis, Rheumatoid - drug therapy ; Arthritis, Rheumatoid - genetics ; Cell Movement - genetics ; Cell Proliferation ; Gentisates - pharmacology ; Humans ; Medicine ; Medicine & Public Health ; MicroRNAs - genetics ; MicroRNAs - metabolism ; Original Article</subject><ispartof>Chinese journal of integrative medicine, 2023-06, Vol.29 (6), p.508-516</ispartof><rights>The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature 2022</rights><rights>2022. The Chinese Journal of Integrated Traditional and Western Medicine Press and Springer-Verlag GmbH Germany, part of Springer Nature.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c344t-9424c4f11806561a3c8bc8d13a56363099a10d03cf5a5d6b39e1cc1ff13362bb3</citedby><cites>FETCH-LOGICAL-c344t-9424c4f11806561a3c8bc8d13a56363099a10d03cf5a5d6b39e1cc1ff13362bb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11655-022-3723-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11655-022-3723-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36251141$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ding, Dou</creatorcontrib><creatorcontrib>Zhang, Qi</creatorcontrib><creatorcontrib>Zeng, Fu-jia</creatorcontrib><creatorcontrib>Cai, Ming-xing</creatorcontrib><creatorcontrib>Gan, Yuan</creatorcontrib><creatorcontrib>Dong, Xiao-jun</creatorcontrib><title>Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis</title><title>Chinese journal of integrative medicine</title><addtitle>Chin. J. Integr. Med</addtitle><addtitle>Chin J Integr Med</addtitle><description>Objective
To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.
Methods
The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.
Results
The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (
P
<0.05 or
P
<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (
P
<0.05 or
P
<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (
P
<0.01).
Conclusion
GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.</description><subject>Arthritis, Rheumatoid - drug therapy</subject><subject>Arthritis, Rheumatoid - genetics</subject><subject>Cell Movement - genetics</subject><subject>Cell Proliferation</subject><subject>Gentisates - pharmacology</subject><subject>Humans</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>MicroRNAs - genetics</subject><subject>MicroRNAs - metabolism</subject><subject>Original Article</subject><issn>1672-0415</issn><issn>1993-0402</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM1OwzAQhC0EoqXwAFxQjlxMvf5LcwwVLUhFoArOluM41FWTFDuR4O1xCXDktLua2dHuh9AlkBsgJJ0GACkEJpRillKG-REaQ5YxTDihx7GXKY09iBE6C2FLiEglEadoxCQVABzG6PnRmo1uXKiTtkqWtulccCbJjSuTtknWG9vXumvjlPtu412Uk1sd7LdauzWGrMBsP13nC0jyDxfO0Umld8Fe_NQJel3cvczv8epp-TDPV9gwzjucccoNrwBmRAoJmplZYWYlMC0kk4xkmQZSEmYqoUUpC5ZZMAaqClg8vijYBF0PuXvfvvc2dKp2wdjdTje27YOiKRWcwyFsgmCwGt-G4G2l9t7V2n8qIOoAUg0gVQSpDiAVjztXP_F9Udvyb-OXXDTQwRCi1LxZr7Zt75v48j-pXxvLerw</recordid><startdate>20230601</startdate><enddate>20230601</enddate><creator>Ding, Dou</creator><creator>Zhang, Qi</creator><creator>Zeng, Fu-jia</creator><creator>Cai, Ming-xing</creator><creator>Gan, Yuan</creator><creator>Dong, Xiao-jun</creator><general>Springer Nature Singapore</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20230601</creationdate><title>Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis</title><author>Ding, Dou ; Zhang, Qi ; Zeng, Fu-jia ; Cai, Ming-xing ; Gan, Yuan ; Dong, Xiao-jun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c344t-9424c4f11806561a3c8bc8d13a56363099a10d03cf5a5d6b39e1cc1ff13362bb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Arthritis, Rheumatoid - drug therapy</topic><topic>Arthritis, Rheumatoid - genetics</topic><topic>Cell Movement - genetics</topic><topic>Cell Proliferation</topic><topic>Gentisates - pharmacology</topic><topic>Humans</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>MicroRNAs - genetics</topic><topic>MicroRNAs - metabolism</topic><topic>Original Article</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ding, Dou</creatorcontrib><creatorcontrib>Zhang, Qi</creatorcontrib><creatorcontrib>Zeng, Fu-jia</creatorcontrib><creatorcontrib>Cai, Ming-xing</creatorcontrib><creatorcontrib>Gan, Yuan</creatorcontrib><creatorcontrib>Dong, Xiao-jun</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Chinese journal of integrative medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ding, Dou</au><au>Zhang, Qi</au><au>Zeng, Fu-jia</au><au>Cai, Ming-xing</au><au>Gan, Yuan</au><au>Dong, Xiao-jun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis</atitle><jtitle>Chinese journal of integrative medicine</jtitle><stitle>Chin. J. Integr. Med</stitle><addtitle>Chin J Integr Med</addtitle><date>2023-06-01</date><risdate>2023</risdate><volume>29</volume><issue>6</issue><spage>508</spage><epage>516</epage><pages>508-516</pages><issn>1672-0415</issn><eissn>1993-0402</eissn><abstract>Objective
To investigate the therapeutic effect of gentisic acid (GA) on rheumatoid arthritis (RA) based on the miR-19b-3p/RAF1 axis.
Methods
The cell counting kit-8 method was used to detect the growth inhibitory effect of different concentrations of GA on MH7A cells, and the drug concentration of GA was determined in the experiment. The quantificational real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-19b-3p and RAF1. RAF1, extracellular regulated protein kinases1/2 (ERK1/2) and phospho-ERK1/2 (p-ERK1/2) were examined by Western blotting. Three methods (dual-luciferase assay, qRT-PCR and Western blot analysis) were used to verify miR-19b-3p targeting RAF1. Flow cytometry was performed to detect MH7A cell apoptosis. Transwell and wound healing assays were used to determine the invasion and migration capacities of MH7A cells.
Results
The growth of MH7A cells was gradually inhibited with increasing GA concentration. When the GA concentration exceeded 80 mmol/L, GA was significantly cytotoxic to MH7A cells, so the half maximal inhibitory concentration of GA for MH7A cells was calculated as 67.019 mmol/L. GA upregulated miR-19b-3p expression, downregulated RAF1 expression, inhibited ERK1/2 phosphorylation, induced MH7A cell apoptosis and suppressed MH7A cell invasion and migration (
P
<0.05 or
P
<0.01). RAF1 was identified as the target of miR-19b-3p and reversed inhibitory effects on miR-19b-3p expression (
P
<0.05 or
P
<0.01). The miR-19b-3p inhibitor upregulated RAF1 expression and ERK1/2 phosphorylation, suppressed MH7A cell apoptosis and induced MH7A cell invasion and migration (
P
<0.01).
Conclusion
GA regulated miR-19b-3p/RAF1 axis to mediate ERK pathway and inhibit the development of RA.</abstract><cop>Singapore</cop><pub>Springer Nature Singapore</pub><pmid>36251141</pmid><doi>10.1007/s11655-022-3723-4</doi><tpages>9</tpages></addata></record> |
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| source | MEDLINE; Alma/SFX Local Collection; SpringerLink Journals - AutoHoldings |
| subjects | Arthritis, Rheumatoid - drug therapy Arthritis, Rheumatoid - genetics Cell Movement - genetics Cell Proliferation Gentisates - pharmacology Humans Medicine Medicine & Public Health MicroRNAs - genetics MicroRNAs - metabolism Original Article |
| title | Mechanism of Gentisic Acid on Rheumatoid Arthritis Based on miR-19b-3p/RAF1 Axis |
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