Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose

Objective To detect expression changes of Sfrp1 in NRK-52E cells after 5-Aza-2'deoxycytidine (5-Aza-CdR) intervention. Methods Sprague-Dawley (SD) rats were divided into normal group (NC group) and diabetic nephropathy group (DN group) randomly. The related biochemical indexes were detected aft...

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Main Author: TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun
Format: Article
Language:zho
Published: Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. 2022-08-01
Series:Jichu yixue yu linchuang
Subjects:
Online Access:http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1194.pdf
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author TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun
author_facet TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun
author_sort TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun
collection DOAJ
description Objective To detect expression changes of Sfrp1 in NRK-52E cells after 5-Aza-2'deoxycytidine (5-Aza-CdR) intervention. Methods Sprague-Dawley (SD) rats were divided into normal group (NC group) and diabetic nephropathy group (DN group) randomly. The related biochemical indexes were detected after successfully modeling[1-2] for 10 weeks;kidney index (KI) was calculated; the expression of Sfrp1 in renal tissue was detected by immunohistochemistry. NRK-52E cells were divided into normal glucose group (NG, 5.5 mmol/L), high glucose group (HG, 30 mmol/L) and high glucose +5-Aza-CdR group(5-Aza-CdR group);the expressions of Sfrp1,Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ were detected by Western blot;the mRNA expression of Sfrp1 was detected by RT-qPCR;pearson test was used to analyze the correlation between Sfrp1 and Dnmt3a,Dnmt3b,col-Ⅲ or col-Ⅳ. Results The level of KI, blood glucose, 24 h urinary protein and creatinine in DN group were all higher than those in NC group.The expression of Sfrp1 in renal tissue of DN group was significantly lower than that of NC group.The results of in vitro experiments showed as follows:the mRNA and protein expression of Sfrp1 were decreased in HG group(P<0.05),while the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in HG group were increased (P<0.05);compared with HG group,the mRNA and protein expression of Sfrp1 was increased in 5-Aza-CdR group(P<0.05), the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in 5-Aza-CdR group were decreased (P<0.05); Sfrp1 was negatively correlated with Dnmt3a (r=-0.937, P<0.05),Dnmt3b (r=-0.965, P<0.05),col-Ⅲ(r=-0.694, P<0.05) and col-Ⅳ(r=-0.888, P<0.05). Conclusions The mechanism through which 5-Aza-2′-deoxycytidine inhibits renal fibrosis is potentially related to the reduction of Dnmt3a and Dnmt3b expression and the increase of Sfrp1 expression.
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spelling doaj.art-5f11e3b85de84978b35180c50199cdb12024-01-05T02:36:08ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252022-08-014281194119910.16352/j.issn.1001-6325.2022.08.1194Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucoseTIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun01. School of Nursing, Sanquan College of Xinxiang Medical University, Xinxiang 453003; ;2. Department of Pathophysiology, Guizhou Medical University,Guiyang 550025; ;3. Guizhou Provincial Key Laboratory of Pathogenesis & Drug Research on Common Chronic Diseases, Guiyang 550025, ChinaObjective To detect expression changes of Sfrp1 in NRK-52E cells after 5-Aza-2'deoxycytidine (5-Aza-CdR) intervention. Methods Sprague-Dawley (SD) rats were divided into normal group (NC group) and diabetic nephropathy group (DN group) randomly. The related biochemical indexes were detected after successfully modeling[1-2] for 10 weeks;kidney index (KI) was calculated; the expression of Sfrp1 in renal tissue was detected by immunohistochemistry. NRK-52E cells were divided into normal glucose group (NG, 5.5 mmol/L), high glucose group (HG, 30 mmol/L) and high glucose +5-Aza-CdR group(5-Aza-CdR group);the expressions of Sfrp1,Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ were detected by Western blot;the mRNA expression of Sfrp1 was detected by RT-qPCR;pearson test was used to analyze the correlation between Sfrp1 and Dnmt3a,Dnmt3b,col-Ⅲ or col-Ⅳ. Results The level of KI, blood glucose, 24 h urinary protein and creatinine in DN group were all higher than those in NC group.The expression of Sfrp1 in renal tissue of DN group was significantly lower than that of NC group.The results of in vitro experiments showed as follows:the mRNA and protein expression of Sfrp1 were decreased in HG group(P<0.05),while the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in HG group were increased (P<0.05);compared with HG group,the mRNA and protein expression of Sfrp1 was increased in 5-Aza-CdR group(P<0.05), the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in 5-Aza-CdR group were decreased (P<0.05); Sfrp1 was negatively correlated with Dnmt3a (r=-0.937, P<0.05),Dnmt3b (r=-0.965, P<0.05),col-Ⅲ(r=-0.694, P<0.05) and col-Ⅳ(r=-0.888, P<0.05). Conclusions The mechanism through which 5-Aza-2′-deoxycytidine inhibits renal fibrosis is potentially related to the reduction of Dnmt3a and Dnmt3b expression and the increase of Sfrp1 expression.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1194.pdf5-aza-2′ deoxycytidine (5-aza-cdr)|dna methyltransferase 3a(dnmt3a)|dna methyltransferase 3b(dnmt3b)|secreted frizzled-related protein 1(sfrp1)|renal fibrosis
spellingShingle TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun
Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
Jichu yixue yu linchuang
5-aza-2′ deoxycytidine (5-aza-cdr)|dna methyltransferase 3a(dnmt3a)|dna methyltransferase 3b(dnmt3b)|secreted frizzled-related protein 1(sfrp1)|renal fibrosis
title Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
title_full Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
title_fullStr Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
title_full_unstemmed Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
title_short Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose
title_sort effect of 5 aza 2 deoxycytidine on the expression of sfrp1 in nrk 52e cells cultured with high glucose
topic 5-aza-2′ deoxycytidine (5-aza-cdr)|dna methyltransferase 3a(dnmt3a)|dna methyltransferase 3b(dnmt3b)|secreted frizzled-related protein 1(sfrp1)|renal fibrosis
url http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-8-1194.pdf
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