Article(id=1228279666851640257, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1228279664221815452, articleNumber=null, orderNo=null, doi=10.12404/j.issn.1671-1815.2409673, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1735401600000, receivedDateStr=2024-12-29, revisedDate=1747584000000, revisedDateStr=2025-05-19, acceptedDate=null, acceptedDateStr=null, onlineDate=1770774292909, onlineDateStr=2026-02-11, pubDate=1754582400000, pubDateStr=2025-08-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770774292909, onlineIssueDateStr=2026-02-11, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770774292909, creator=13701087609, updateTime=1770774292909, updator=13701087609, issue=Issue{id=1228279664221815452, tenantId=1146029695717560320, journalId=1146123166801305609, year='2025', volume='25', issue='22', pageStart='9211', pageEnd='9648', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1770774292283, creator=13701087609, updateTime=1770777611996, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1228293588207992892, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1228279664221815452, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1228293588207992893, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1228279664221815452, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=9305, endPage=9311, ext={EN=ArticleExt(id=1228279669057844172, articleId=1228279666851640257, tenantId=1146029695717560320, journalId=1146123166801305609, language=EN, title=Effect of lncRNA Rpph1 on AMPK/Nrf2 Signal Pathway and Cytopyrosis in Diabetes Nephropathy, columnId=1228279667216544709, journalTitle=Science Technology and Engineering, columnName=Papers·Medicine, runingTitle=null, highlight=null, articleAbstract=

To investigate the mechanism of lncRNA (long chain non coding RNA) Rpph1 activating cytopyrosis through AMP-AMPK(activated protein kinase)/Nrf2(nuclear factor E2 related factor 2) signaling pathway, then to promote podocyte injury in DN (diabetes nephropathy). HGPC(Human glomerular podocytes) were cultured in vitro and randomly divided into control group, model group, lncRNA Rpph1 over-expression group, low-expression group, and empty vector group. HGPC were incubated with 5 mmol/L D-glucose as control group, while the other three groups were incubated with 30 mmol/L D-glucose to establish DN model. Liposome transfection method was used to co-incubate stable plasmids carrying Rpph1 over-expression, low-expression, and empty vector with HGPC. qRT-PCR was used to detect lncRNA Rpph1 expression, Western blot was used to detect p-AMPK/AMPK and Nrf2 proteins, as well as the expression levels of cytopyrosis related proteins including NLRP3(Nod like receptor thermal domain associated protein 3), caspase-1, and GSDMD-N. MTT assay was used to detect cell survival rate. Flow cytometry was used to detecte apoptosis rate. Compared with control group, the expression level of lncRNA Rpph1 in model group significantly increased (P<0.05). The expression levels of p-AMPK/AMPK, Nrf2, NLRP3, caspase-1, and GSDMD-N proteins significantly increased in model group (P<0.05). The survival rate of model group cells significantly reduced, while apoptosis rate increased in model group (P<0.05). Compared with model group and empty vector group, lncRNA Rpph1, p-AMPK/AMPK, Nrf2,NLRP3, caspase-1, and GSDMD-N proteins in lncRNA Rpph1 over-expression group significantly increased, and cell survival rate significantly reduced, apoptosis rate increased (P<0.05). The expression levels of lncRNA Rpph1, p-AMPK/AMPK, Nrf2, NLRP3, caspase-1, and GSDMD-N proteins significantly decreased in lncRNA Rpph1 low-expression group, and cell survival rate significantly increased, apoptosis rate reduced (P<0.05). In all, High expression of lncRNA Rpph1 in DN may activate cytopyrosis and promote podocyte injury by AMPK/Nrf2 signaling pathway.

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探讨长链非编码RNA(lncRNA)Rpph1促进糖尿病肾病(diabetes nephropathy,DN)足细胞损伤,是否与腺苷酸激活蛋白激酶(AMP-activated protein kinase,AMPK)/核因子E2相关因子2(Nrf2)信号通路以及细胞焦亡和凋亡有关。体外培养人肾小球足细胞(Human glomerular podocytes,HGPC)随机分为对照组、模型组、lncRNA Rpph1过表达组、低表达组和空载体组,5 mmol/L的D-葡萄糖孵育HGPC为对照组,其他三组采用30 mmol/L的D-葡萄糖孵育细胞建立DN模型。脂质体转染法将携带lncRNA Rpph1过表达、低表达与空载体的稳定质粒与HGPC共孵育。qRT-PCR检测lncRNA Rpph1表达,Western blot检测p-AMPK/AMPK和Nrf2蛋白,以及细胞焦亡相关蛋白包括Nod样受体热蛋白结构域相关蛋白3(NLRP3)、胱天蛋白酶-1(caspase-1)和GSDMD-N的表达量,MTT法检测细胞存活率,流式细胞术检测凋亡率。与对照组相比,模型组lncRNA Rpph1表达量显著增加(P<0.05)。模型组p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加(P<0.05)。模型组细胞存活率显著减少,凋亡率增加(P<0.05)。与模型组和空载体组相比,lncRNA Rpph1过表达组lncRNA Rpph1、p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加,细胞存活率显著减少,凋亡率增加(P<0.05);lncRNA Rpph1低表达组lncRNA Rpph1、p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著下降,细胞存活率显著增多,凋亡率下降(P<0.05)。DN中lncRNA Rpph1高表达可以促进足细胞损伤,可能通过AMPK/Nrf2信号通路激活细胞焦亡和凋亡有关。

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古丽鲜·吐尔洪(1983—),女,维吾尔族,新疆乌鲁木齐人,硕士,副主任医师。研究方向:慢性肾小球疾病。E-mail:

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古丽鲜·吐尔洪(1983—),女,维吾尔族,新疆乌鲁木齐人,硕士,副主任医师。研究方向:慢性肾小球疾病。E-mail:

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古丽鲜·吐尔洪(1983—),女,维吾尔族,新疆乌鲁木齐人,硕士,副主任医师。研究方向:慢性肾小球疾病。E-mail:

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Chinese Journal of Clinical Pharmacology, 2023, 39(22): 3271-3275., articleTitle=Effect of crocetin on oxidative stress and pyroptosis of HK-2 cells induced by high glucose in diabetes nephropathy, refAbstract=null), Reference(id=1228369784044192302, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2024, volume=27, issue=21, pageStart=2617, pageEnd=2622, url=null, language=null, rfNumber=[21], rfOrder=35, authorNames=李佳武, 秦凤, 宋生琴, journalName=中国全科医学, refType=null, unstructuredReference=李佳武, 秦凤, 宋生琴, 等. 基于NLRP3/IL-1β/TGF-β1通路探讨低氧环境下红景天苷对糖尿病肾病大鼠足细胞焦亡损伤的拮抗效应[J]. 中国全科医学, 2024, 27(21): 2617-2622., articleTitle=基于NLRP3/IL-1β/TGF-β1通路探讨低氧环境下红景天苷对糖尿病肾病大鼠足细胞焦亡损伤的拮抗效应, refAbstract=null), Reference(id=1228369784144855602, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2024, volume=27, issue=21, pageStart=2617, pageEnd=2622, url=null, language=null, rfNumber=[21], rfOrder=36, authorNames=Li Jiawu, Qin Feng, Song Shengqin, journalName=China General Medicine, refType=null, unstructuredReference=Li Jiawu, Qin Feng, Song Shengqin, et al. Based on NLRP3/IL-1β/TGF-β1 pathway, explore the antagonistic effect of salidroside on podocyte scorch injury in diabetes nephropathy rats under hypoxic environment[J]. China General Medicine, 2024, 27 (21): 2617-2622., articleTitle=Based on NLRP3/IL-1β/TGF-β1 pathway, explore the antagonistic effect of salidroside on podocyte scorch injury in diabetes nephropathy rats under hypoxic environment, refAbstract=null), Reference(id=1228369784224547384, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2023, volume=34, issue=10, pageStart=1478, pageEnd=1486, url=null, language=null, rfNumber=[22], rfOrder=37, authorNames=吴思宇, 顾惠贤, 张文祥, journalName=中药新药与临床药理, refType=null, unstructuredReference=吴思宇, 顾惠贤, 张文祥, 等. 非编码RNA调控糖尿病肾病细胞焦亡的研究进展[J]. 中药新药与临床药理, 2023, 34(10): 1478-1486., articleTitle=非编码RNA调控糖尿病肾病细胞焦亡的研究进展, refAbstract=null), Reference(id=1228369784346182205, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2023, volume=34, issue=10, pageStart=1478, pageEnd=1486, url=null, language=null, rfNumber=[22], rfOrder=38, authorNames=Wu Siyu, Gu Huixian, Zhang Wenxiang, journalName=New Traditional Chinese Medicine and Clinical Pharmacology, refType=null, unstructuredReference=Wu Siyu, Gu Huixian, Zhang Wenxiang, et al. Research progress of non coding RNA regulating pyroptosis in diabetes nephropathy[J]. New Traditional Chinese Medicine and Clinical Pharmacology, 2023, 34(10): 1478-1486., articleTitle=Research progress of non coding RNA regulating pyroptosis in diabetes nephropathy, refAbstract=null), Reference(id=1228369784467817027, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2023, volume=39, issue=1, pageStart=12, pageEnd=20, url=null, language=null, rfNumber=[23], rfOrder=39, authorNames=汪乐新, 刘超, 马天龙, journalName=实用医学杂志, refType=null, unstructuredReference=汪乐新, 刘超, 马天龙, 等. LncRNA H19调控肾小球足细胞焦亡中作用[J]. 实用医学杂志, 2023, 39(1): 12-20., articleTitle=LncRNA H19调控肾小球足细胞焦亡中作用, refAbstract=null), Reference(id=1228369784551703112, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, doi=null, pmid=null, pmcid=null, year=2023, volume=39, issue=1, pageStart=12, pageEnd=20, url=null, language=null, rfNumber=[23], rfOrder=40, authorNames=Wang Lexin, Liu Chao, Ma Tianlong, journalName=Journal of Practical Medicine, refType=null, unstructuredReference=Wang Lexin, Liu Chao, Ma Tianlong, et al. The role of LncRNA H19 in regulating glomerular podocyte pyroptosis[J]. Journal of Practical Medicine, 2023, 39(1): 12-20., articleTitle=The role of LncRNA H19 in regulating glomerular podocyte pyroptosis, refAbstract=null)], funds=[Fund(id=1228369778654511467, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, awardId=2021D03023, language=CN, fundingSource=新疆少数民族科技人才特殊培养计划(2021D03023), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1228369771033461759, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, xref=null, ext=[AuthorCompanyExt(id=1228369771037656064, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, companyId=1228369771033461759, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Nephrology Department of the Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830028, China), AuthorCompanyExt(id=1228369771046043649, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, companyId=1228369771033461759, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=新疆医科大学第二附属医院肾内科, 乌鲁木齐 830028)])], figs=[ArticleFig(id=1228369774686699700, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.1, caption=qRT-PCR detection of lncRNA Rpph1 expression levels in podocytes between the control group and the model group, figureFileSmall=8w3joD3bkLatrTwn/0H04w==, figureFileBig=PvEWwkjTmU3m2qHwDnAMtg==, tableContent=null), ArticleFig(id=1228369774787363001, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图1, caption=对照组与模型组足细胞lncRNA Rpph1表达量

**表示P<0.01

, figureFileSmall=8w3joD3bkLatrTwn/0H04w==, figureFileBig=PvEWwkjTmU3m2qHwDnAMtg==, tableContent=null), ArticleFig(id=1228369774917386439, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.2, caption=qRT-PCR detection of lncRNA Rpph1 expression levels in podocytes of each group, figureFileSmall=QsBc7lq0Lxjh94Ix6W/pLA==, figureFileBig=3Iq0wSWLU8nDT5Y5DYnoFw==, tableContent=null), ArticleFig(id=1228369775009661134, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图2, caption=qRT-PCR检测各组足细胞lncRNA Rpph1表达量

**表示P<0.01

, figureFileSmall=QsBc7lq0Lxjh94Ix6W/pLA==, figureFileBig=3Iq0wSWLU8nDT5Y5DYnoFw==, tableContent=null), ArticleFig(id=1228369775139684566, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.3, caption=AMPK/Nrf2 signaling pathway protein expression levels in podocytes of each group by Western blot, figureFileSmall=pCoDc1F5EnL1LNCYppYUVA==, figureFileBig=EVK6wfSX5vfzKQvCWI54Fw==, tableContent=null), ArticleFig(id=1228369775257125088, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图3, caption=Western blot检测各组足细胞AMPK/Nrf2信号通路蛋白表达量

**表示P<0.01

, figureFileSmall=pCoDc1F5EnL1LNCYppYUVA==, figureFileBig=EVK6wfSX5vfzKQvCWI54Fw==, tableContent=null), ArticleFig(id=1228369775370371304, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.4, caption=Statistical analysis results of AMPK signaling pathway protein in each group, figureFileSmall=DLi/WmoNE1khlWenj1rSgA==, figureFileBig=IOnnHVygmdMsu5qBFwqsTQ==, tableContent=null), ArticleFig(id=1228369775475228912, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图4, caption=各组足细胞AMPK通路蛋白统计分析结果

**表示P<0.01

, figureFileSmall=DLi/WmoNE1khlWenj1rSgA==, figureFileBig=IOnnHVygmdMsu5qBFwqsTQ==, tableContent=null), ArticleFig(id=1228369775592669431, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.5, caption=Statistical analysis results of Nrf2 signaling pathway protein in each group, figureFileSmall=+JQU7htMSKjnymeb8cTgLw==, figureFileBig=hKqG9K3uiXxwcByF3otGSg==, tableContent=null), ArticleFig(id=1228369775697527040, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图5, caption=各组足细胞Nrf2信号通路蛋白统计分析结果

**表示P<0.01

, figureFileSmall=+JQU7htMSKjnymeb8cTgLw==, figureFileBig=hKqG9K3uiXxwcByF3otGSg==, tableContent=null), ArticleFig(id=1228369775777218822, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.6, caption=Pyroptosis protein expression in podocyte cells of each group by Western blot, figureFileSmall=1RSdLJ9r73UDy4+Irt9PUA==, figureFileBig=yqCZJuakIVXPggJoT01G9A==, tableContent=null), ArticleFig(id=1228369775865299211, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图6, caption=Western blot检测各组足细胞细胞焦亡蛋白表达量

**表示P<0.01

, figureFileSmall=1RSdLJ9r73UDy4+Irt9PUA==, figureFileBig=yqCZJuakIVXPggJoT01G9A==, tableContent=null), ArticleFig(id=1228369775957573909, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.7, caption=Statistical analysis results of NLRP3 protein in each group, figureFileSmall=aoORGCGuEFhsslOpFBcQEA==, figureFileBig=8c9ZPQaJmvHd3Fq5BtFeEw==, tableContent=null), ArticleFig(id=1228369776054042907, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图7, caption=各组足细胞NLRP3蛋白统计分析结果

**表示P<0.01

, figureFileSmall=aoORGCGuEFhsslOpFBcQEA==, figureFileBig=8c9ZPQaJmvHd3Fq5BtFeEw==, tableContent=null), ArticleFig(id=1228369776133734689, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.8, caption=Statistical analysis results of caspase-1 protein in each group, figureFileSmall=aWokM6ppji89AkWJ4Mp+Jw==, figureFileBig=0f7nnXffsVneL0PAPgfeNw==, tableContent=null), ArticleFig(id=1228369776221815081, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图8, caption=各组足细胞caspase-1蛋白统计分析结果

**表示P<0.01

, figureFileSmall=aWokM6ppji89AkWJ4Mp+Jw==, figureFileBig=0f7nnXffsVneL0PAPgfeNw==, tableContent=null), ArticleFig(id=1228369776305701170, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.9, caption=Statistical analysis results of GSDMD-N protein in each group, figureFileSmall=AhWJQ5k3oNkN8j1AnwFZQA==, figureFileBig=2VJHrr0BPMUA5aQ9VtO+5g==, tableContent=null), ArticleFig(id=1228369776410558777, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图9, caption=各组足细胞GSDMD-N蛋白统计分析结果

**表示P<0.01

, figureFileSmall=AhWJQ5k3oNkN8j1AnwFZQA==, figureFileBig=2VJHrr0BPMUA5aQ9VtO+5g==, tableContent=null), ArticleFig(id=1228369776527999297, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.10, caption=The survival of podocytes in each group by MTT, figureFileSmall=ZqPIcUMLQafkP+RYX6UGPg==, figureFileBig=VyOhh8T7Z6IhOIxpmNrI9g==, tableContent=null), ArticleFig(id=1228369776632856901, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图10, caption=MTT检测各组足细胞存活

**表示P<0.01

, figureFileSmall=ZqPIcUMLQafkP+RYX6UGPg==, figureFileBig=VyOhh8T7Z6IhOIxpmNrI9g==, tableContent=null), ArticleFig(id=1228369776825794893, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.11, caption=Podocyte apoptosis in each group, figureFileSmall=/jihTpsrKkm5uR1BVRzlHg==, figureFileBig=R4/UZ7QNZMLlyV/2b/gYkw==, tableContent=null), ArticleFig(id=1228369776909680980, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图11, caption=各组足细胞凋亡情况

**表示P<0.01

, figureFileSmall=/jihTpsrKkm5uR1BVRzlHg==, figureFileBig=R4/UZ7QNZMLlyV/2b/gYkw==, tableContent=null), ArticleFig(id=1228369778297995613, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=EN, label=Fig.12, caption=The comparison of apoptosis rate in each group, figureFileSmall=fT+GkbcYJw0sauFsFMw2KA==, figureFileBig=r0DS/K/07PAiHte6tnx7nw==, tableContent=null), ArticleFig(id=1228369778448990560, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1228279666851640257, language=CN, label=图12, caption=各组凋亡率比较

**表示P<0.01

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医药、卫生lncRNA Rpph1对糖尿病肾病足细胞AMPK/Nrf2通路、焦亡的影响
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古丽鲜·吐尔洪 , 姑丽孜巴·塔衣尔
科学技术与工程 | 论文·医药、卫生 2025,25(22): 9305-9311
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科学技术与工程 | 论文·医药、卫生 2025, 25(22): 9305-9311
医药、卫生lncRNA Rpph1对糖尿病肾病足细胞AMPK/Nrf2通路、焦亡的影响
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古丽鲜·吐尔洪 , 姑丽孜巴·塔衣尔
作者信息
  • 新疆医科大学第二附属医院肾内科, 乌鲁木齐 830028
  • 古丽鲜·吐尔洪(1983—),女,维吾尔族,新疆乌鲁木齐人,硕士,副主任医师。研究方向:慢性肾小球疾病。E-mail:

Effect of lncRNA Rpph1 on AMPK/Nrf2 Signal Pathway and Cytopyrosis in Diabetes Nephropathy
GULIXIAN·Turhong , GULIZIBA·Tayier
Affiliations
  • Nephrology Department of the Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830028, China
出版时间: 2025-08-08 doi: 10.12404/j.issn.1671-1815.2409673
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探讨长链非编码RNA(lncRNA)Rpph1促进糖尿病肾病(diabetes nephropathy,DN)足细胞损伤,是否与腺苷酸激活蛋白激酶(AMP-activated protein kinase,AMPK)/核因子E2相关因子2(Nrf2)信号通路以及细胞焦亡和凋亡有关。体外培养人肾小球足细胞(Human glomerular podocytes,HGPC)随机分为对照组、模型组、lncRNA Rpph1过表达组、低表达组和空载体组,5 mmol/L的D-葡萄糖孵育HGPC为对照组,其他三组采用30 mmol/L的D-葡萄糖孵育细胞建立DN模型。脂质体转染法将携带lncRNA Rpph1过表达、低表达与空载体的稳定质粒与HGPC共孵育。qRT-PCR检测lncRNA Rpph1表达,Western blot检测p-AMPK/AMPK和Nrf2蛋白,以及细胞焦亡相关蛋白包括Nod样受体热蛋白结构域相关蛋白3(NLRP3)、胱天蛋白酶-1(caspase-1)和GSDMD-N的表达量,MTT法检测细胞存活率,流式细胞术检测凋亡率。与对照组相比,模型组lncRNA Rpph1表达量显著增加(P<0.05)。模型组p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加(P<0.05)。模型组细胞存活率显著减少,凋亡率增加(P<0.05)。与模型组和空载体组相比,lncRNA Rpph1过表达组lncRNA Rpph1、p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加,细胞存活率显著减少,凋亡率增加(P<0.05);lncRNA Rpph1低表达组lncRNA Rpph1、p-AMPK/AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N蛋白表达量显著下降,细胞存活率显著增多,凋亡率下降(P<0.05)。DN中lncRNA Rpph1高表达可以促进足细胞损伤,可能通过AMPK/Nrf2信号通路激活细胞焦亡和凋亡有关。

糖尿病肾病  /  足细胞  /  长链非编码RNA Rpph1  /  腺苷酸激活蛋白激酶  /  核因子E2相关因子2  /  细胞焦亡

To investigate the mechanism of lncRNA (long chain non coding RNA) Rpph1 activating cytopyrosis through AMP-AMPK(activated protein kinase)/Nrf2(nuclear factor E2 related factor 2) signaling pathway, then to promote podocyte injury in DN (diabetes nephropathy). HGPC(Human glomerular podocytes) were cultured in vitro and randomly divided into control group, model group, lncRNA Rpph1 over-expression group, low-expression group, and empty vector group. HGPC were incubated with 5 mmol/L D-glucose as control group, while the other three groups were incubated with 30 mmol/L D-glucose to establish DN model. Liposome transfection method was used to co-incubate stable plasmids carrying Rpph1 over-expression, low-expression, and empty vector with HGPC. qRT-PCR was used to detect lncRNA Rpph1 expression, Western blot was used to detect p-AMPK/AMPK and Nrf2 proteins, as well as the expression levels of cytopyrosis related proteins including NLRP3(Nod like receptor thermal domain associated protein 3), caspase-1, and GSDMD-N. MTT assay was used to detect cell survival rate. Flow cytometry was used to detecte apoptosis rate. Compared with control group, the expression level of lncRNA Rpph1 in model group significantly increased (P<0.05). The expression levels of p-AMPK/AMPK, Nrf2, NLRP3, caspase-1, and GSDMD-N proteins significantly increased in model group (P<0.05). The survival rate of model group cells significantly reduced, while apoptosis rate increased in model group (P<0.05). Compared with model group and empty vector group, lncRNA Rpph1, p-AMPK/AMPK, Nrf2,NLRP3, caspase-1, and GSDMD-N proteins in lncRNA Rpph1 over-expression group significantly increased, and cell survival rate significantly reduced, apoptosis rate increased (P<0.05). The expression levels of lncRNA Rpph1, p-AMPK/AMPK, Nrf2, NLRP3, caspase-1, and GSDMD-N proteins significantly decreased in lncRNA Rpph1 low-expression group, and cell survival rate significantly increased, apoptosis rate reduced (P<0.05). In all, High expression of lncRNA Rpph1 in DN may activate cytopyrosis and promote podocyte injury by AMPK/Nrf2 signaling pathway.

diabetes nephropathy  /  podocyte  /  long non coding RNA Rpph1  /  AMP-activated protein kinase  /  nuclear factor E2 related factor 2  /  cytopyrosis
古丽鲜·吐尔洪, 姑丽孜巴·塔衣尔. 医药、卫生lncRNA Rpph1对糖尿病肾病足细胞AMPK/Nrf2通路、焦亡的影响. 科学技术与工程, 2025 , 25 (22) : 9305 -9311 . DOI: 10.12404/j.issn.1671-1815.2409673
GULIXIAN·Turhong, GULIZIBA·Tayier. Effect of lncRNA Rpph1 on AMPK/Nrf2 Signal Pathway and Cytopyrosis in Diabetes Nephropathy[J]. Science Technology and Engineering, 2025 , 25 (22) : 9305 -9311 . DOI: 10.12404/j.issn.1671-1815.2409673
糖尿病肾病(diabetes nephropathy,DN)糖尿病肾病(diabetic kidney disease,DKD)是指由糖尿病引起的慢性肾脏疾病,现已成为慢性肾脏病和终末期肾病的主要原因。发病机制,涉及炎症反应、氧化应激、细胞凋亡、高糖毒性等[1]。足细胞、肾小球内皮细胞及肾小球基底膜共同构成了肾小球滤过屏障,调控肾小球滤过功能。足细胞损伤、脱落和凋亡等是DKD最具特征的病理变化[2],因此探索足细胞损伤机制至关重要。研究发现,长链非编码RNA(long non coding RNA,lncRNA)在DN的发病机制中发挥重要作用,可以影响下游靶基因和效应蛋白的功能表达,进而加重DN病情和影响预后。lncRNA Rpph1是一类重要的lncRNA,既往研究发现其在恶性肿瘤[3]、糖尿病[4]、肾小管上皮细胞损伤[5]等疾病中异常表达,可能参与了疾病的发生。
但是,中外关于lncRNA Rpph1在DN中的确切发病机制仍不是十分清楚。细胞焦亡是一种细胞程序性死亡方式,已经被证实在DN发病机制中同样发挥重要作用[6-7]。lncRNA Rpph1是否通过调控细胞焦亡进而影响DN足细胞的损伤还没有统一认识。腺苷酸激活蛋白激酶(AMP-activated protein kinase,AMPK)/核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)信号通路在调控细胞增殖、凋亡、炎症反应等病理过程中发挥重要作用[8-9]。基于此,现利用高糖刺激人肾小球足细胞建立DN模型,探讨lncRNA Rpph1促进DN足细胞损伤的相关机制,是否与AMPK/Nrf2信号通路以及细胞焦亡和凋亡有关。
体外培养人肾小球足细胞(human glomerular podocytes,HGPC)购自上海生工细胞实验中心,常规复苏后在DMEM-F12培养基(含10%胎牛血清和1%青-链霉素)中,于37 ℃ 5% CO2培养箱中培养,隔天换液至细胞体积达到85%后,胰酶消化终止反应,PBS洗涤重悬细胞浓度为1×106/mL,分装备用。
Lipofectamine 2000转染试剂购自赛默飞世尔科技(中国)有限公司,逆转录试剂盒和PCR试剂盒购自美国R&D公司,BCA蛋白定量试剂盒购自美国Sigma公司,鼠抗人p-AMPK、AMPK、Nrf2、NLRP3、caspase-1和GSDMD-N抗体一抗及对应二抗购自江苏碧云天科技有限公司,MTT试剂盒购自翌圣生物科技(上海)股份有限公司。
HGPC细胞随机分为对照组、模型组、lncRNA Rpph1过表达组、低表达组和空载体组,5 mmol/L的D-葡萄糖孵育HGPC为对照组,其他3组采用30 mmol/L的D-葡萄糖孵育细胞建立DN模型。
脂质体转染法将携带lncRNA Rpph1过表达、低表达与空载体的稳定质粒与HGPC共孵育24 h。实时荧光定量PCR(qRT-PCR)检测lncRNA Rpph1表达量,鉴定转染效率。
Trizol试剂提取细胞RNA,逆转录合成cDNA后测量纯度和浓度,设计引物序列lncRNA Rpph1:上游5’-ACTGGGTGTGATGCCTCTCAAG-3’,下游5’-GGAACCTGAACCCCTGCTGTG-3’;内参β-actin:上游5’CTGGTATCGTGGAAGGACTC-3’,下游5’-GTAGAGGCAGGGATGATGTCT-3’。根据试剂盒说明书配制反应体系包括10×PCR缓冲液、MgCl2、dNTPs、Taq 酶、cDNA和反应水,总体积30 μL。反应条件为94 ℃预变性5 min;94 ℃ 45 s、60 ℃ 45 s、72 ℃ 45 s,共循环 40次;72 ℃延伸10 min结束。构建扩增曲线和熔解曲线,结果以2-△△Ct表示。
蛋白包括p-AMPK/AMPK和Nrf2蛋白,以及细胞焦亡相关蛋白包括Nod样受体热蛋白结构域相关蛋白3(nod like receptor thermal domain associated protein 3,NLRP3)、胱天蛋白酶-1(caspase-1)和GSDMD-N的表达量。RIPA裂解液提取蛋白,对蛋白进行定量、电泳分离,转PVDF膜后室温封闭2 h。滴加鼠抗人单克隆抗体一抗(稀释浓度为1∶1 000),4 ℃孵育过夜,洗涤后加入辣根酶标记羊抗兔二抗(稀释浓度为1∶500)室温下孵育4 h。PBS洗涤,ECL显色,Lab Works 4.5凝胶成像软件行半定量分析,以目的蛋白与内参GAPDH条带的灰度值比值表示。每组设置3个复孔,结果取平均值。
各组处理24 h后分别加入 MTT 每孔 20 μL,继续孵育 4 h后弃上清液,加 DMSO 每孔 200 μL,振荡 10 min,置于波长 570 nm 处测定吸光度(A)值。细胞存活率=A 处理组/A 对照组×100%。每组设置6个复孔,结果取平均值。
各组细胞培养24 h后用预冷PBS 冲洗,加入 100 μL 1×结合缓冲液重悬,然后加入 5 μL Annexin V-FITC 和 PI 染液充分混匀,室温遮光染色 15 min,流式细胞仪检测细胞凋亡率。
采用SPSS 23.0统计软件对计量资料(均数±标准差)多组间比较采用单因素ANOVA分析,然后两两组间再比较采用LSD-t法检验;P<0.05表示差异有统计学意义。
与对照组相比,模型组lncRNA Rpph1表达量显著增加(P<0.05),提示DN中足细胞lncRNA Rpph1表达量上调。与空载体组相比,lncRNA Rpph1过表达组lncRNA Rpph1表达量显著增加,lncRNA Rpph1低表达组lncRNA Rpph1表达量显著下降(P<0.05),提示转染成功,如图1图2所示。
与对照组相比,模型组p-AMPK/AMPK和Nrf2蛋白表达量显著增加(P<0.05),提示AMPK/Nrf2信号通路激活参与了DN足细胞的损伤。与模型组和空载体组相比,lncRNA Rpph1过表达组p-AMPK/AMPK和Nrf2蛋白表达量显著增加,lncRNA Rpph1低表达组p-AMPK/AMPK和Nrf2蛋白表达量显著下降(P<0.05),提示靶向干预lncRNA Rpph1表达可以正向调控AMPK/Nrf2信号通路的活性,如图3~图5所示。
与对照组相比,模型组的NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加(P<0.05),提示细胞焦亡参与了DN足细胞的损伤。与模型组和空载体组相比,lncRNA Rpph1过表达组NLRP3、caspase-1和GSDMD-N蛋白表达量显著增加,lncRNA Rpph1低表达组NLRP3、caspase-1和GSDMD-N蛋白表达量显著下降(P<0.05),提示靶向干预lncRNA Rpph1表达可以影响细胞焦亡的活性,如图6~图9所示。
与对照组相比,模型组的细胞存活率显著减少,凋亡率显著增高(P<0.05),提示DN可导致足细胞损伤。与模型组和空载体组相比,lncRNA Rpph1过表达组的细胞存活率显著减少,凋亡率显著增加;lncRNA Rpph1低表达组的细胞存活率显著增多,凋亡率显著降低(P<0.05),提示靶向干预lncRNA Rpph1表达可以影响DN足细胞损伤,如图10~图12所示。
DN是糖尿病最常见的微血管并发症之一,发病机制复杂,涉及多种细胞信号通路和分子调控网络。足细胞是一种高度分化的肾脏实质细胞,是肾小球滤过屏障的关键组成部分,当足细胞受到损伤时,肾小球滤过屏障受损,蛋白尿生成,最终导致肾小球滤过率的下降和终末期肾病。然而足细胞的增殖能力有限,一旦出现损伤则无法通过自我调节来弥补。
LncRNA是缺乏编码蛋白能力的内源性分子,其在肿瘤、心脑血管疾病和神经退行性疾病中具有重要调节作用。越来越多的研究证明LncRNA可直接或间接参与 DKD 的发生、发展。lncRNA Rpph1作为一种重要的调控分子,表达水平在DN患者肾组织中显著上调,提示lncRNA Rpph1可能参与了该疾病的发生、发展[10]。本文研究中模型组lncRNA Rpph1表达量显著增加,同样提示DN中足细胞lncRNA Rpph1表达量上调。lncRNA Rpph1是一种由RNA聚合酶III转录的lncRNA,主要定位于细胞核内,参与多种生理病理过程的调控。正常生理条件下,lncRNA Rpph1主要调控细胞的增殖、分化和凋亡等基本生命活动。lncRNA Rpph1通过与特定的转录因子或表观遗传调控因子结合,影响下游基因的表达。例如,lncRNA Rpph1可以招募EZH2等表观遗传调控蛋白,下调肿瘤抑制基因的表达,从而促进肿瘤细胞的增殖和侵袭[11]。lncRNA Rpph1还可以通过竞争性结合microRNA方式,调控靶基因的表达,参与机体的免疫调节和炎症反应[12-13]。lncRNA Rpph1作为一个多功能调控因子,表达水平异常可能导致多种疾病的发生与发展[14]
在DN发病过程中,lncRNA Rpph1表达水平显著上调,可能由于高糖环境下lncRNA Rpph1转录被激活[15]。并且,lncRNA Rpph1通过多种机制参与DN的发病过程。lncRNA Rpph1可以抑制肾小管细胞的增殖,促进细胞凋亡,加重肾脏功能的损害;lncRNA Rpph1可以诱导细胞外基质的过度沉积,加重肾小球硬化;lncRNA Rpph1可以激活炎症信号通路,增强肾脏的炎症反应,加重肾脏损伤;lncRNA Rpph1可以通过调控DNA甲基转移酶的表达,改变肾小管细胞的表观遗传状态,影响细胞的增殖和分化[16-17]。因此,推测lncRNA Rpph1在DN发病机制中发挥重要的作用,可作为疾病的潜在治疗靶点。进一步通过靶向干预lncRNA Rpph1表达发现,可以正向调控AMPK/Nrf2信号通路的活性。王静等[18]指出,AMPK/Nrf2信号通路参与了DN的发生以及药物治疗过程。何亚萍等[19]也指出,Nrf2参与了DN的发病机制。
本文研究还显示,靶向干预lncRNA Rpph1表达可以影响细胞焦亡的活性和足细胞的存活率和凋亡率。姜莉莉等[20]指出,系膜细胞的氧化应激和焦亡参与了DN的发生。李佳武等[21]指出,NLRP3/IL-1β/TGF-β1 通路参与了DN中足细胞的焦亡损伤过程。研究发现,lncRNA Rpph1可以通过多种方式参与调控DN足细胞的焦亡过程[22-23]。lncRNA Rpph1可以抑制Sirt1表达,激活TGF-β/Smad通路,促进肾小球系膜细胞和足细胞的增殖和细胞外基质的沉积。lncRNA Rpph1可以竞争性结合miR-29b,降低对Col4a1、Col4a2等基因的抑制作用,加重肾脏纤维化。lncRNA Rpph1可以招募EZH2抑制Klotho基因的表达,加重肾脏功能的损害。lncRNA Rpph1还可以通过调控NF-κB信号通路,增强肾脏的炎症反应。lncRNA Rpph1作为一个“主控”因子,通过调控多条信号通路参与DN的发病机制。
DN中lncRNA Rpph1高表达可以促进足细胞损伤,可能通过AMPK/Nrf2信号通路调控细胞焦亡。干预lncRNA Rpph1有望成为临床诊治DN的新型靶点。
  • 新疆少数民族科技人才特殊培养计划(2021D03023)
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2025年第25卷第22期
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doi: 10.12404/j.issn.1671-1815.2409673
  • 接收时间:2024-12-29
  • 首发时间:2026-02-11
  • 出版时间:2025-08-08
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  • 收稿日期:2024-12-29
  • 修回日期:2025-05-19
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新疆少数民族科技人才特殊培养计划(2021D03023)
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    新疆医科大学第二附属医院肾内科, 乌鲁木齐 830028
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2种不同金属材料的力学参数

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属数
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genus
种数
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species
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Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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