Suppression of XBP1S mediates high glucose-induced oxidative stress and extracellular matrix synthesis in renal mesangial cell and kidney of diabetic rats.
Suppression of XBP1S mediates high glucose-induced oxidative stress and extracellular matrix synthesis in renal mesangial cell and kidney of diabetic rats.
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XBP1S 的抑制介导糖尿病大鼠肾系膜细胞和肾脏中高糖诱导的氧化应激和细胞外基质合成
DOI：
10.1371/journal.pone.0056124
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lu L
中科院分区：
文献类型：
--
作者：
Shao D;Liu J;Ni J;Wang Z;Shen Y;Zhou L;Huang Y;Wang J;Xue H;Zhang W;Lu L
近年来的研究表明，内质网（ER）应激参与了包括糖尿病肾病（DN）在内的多种病理过程。X-box结合蛋白1（X-box binding protein 1，XBP 1）作为内质网应激的重要介质，具有抗氧化应激的作用。在这项研究中，我们研究了剪接的XBP 1（XBP 1 S），XBP 1的主要活性形式，对高糖（HG）诱导的活性氧（ROS）的产生和细胞外基质（ECM）的合成在培养的肾系膜细胞（MCs）和STZ诱导的糖尿病大鼠肾皮质的影响。真实的时间PCR和蛋白质印迹分别用于评估mRNA和蛋白质水平。用携带XBP 1 S基因的重组腺病毒载体（Ad-XBP 1 S）转染大肠杆菌，使XBP 1 S表达上调。XBP 1 S siRNA用于敲低XBP 1 S表达。二氢乙锭（DHE）荧光探针法检测细胞内活性氧水平。结果显示，HG处理后，MCs中XBP 1 S蛋白和mRNA水平明显降低，未剪接的XBP 1（XBP 1U）蛋白和mRNA水平无明显变化。同时，ROS的产生，IV型胶原和纤维连接蛋白的表达增加。氯化二苯碘（DPI）是一种NADPH氧化酶抑制剂，可阻止HG诱导的活性氧增加以及IV型胶原和纤连蛋白表达。Ad-XBP 1 S转染逆转HG诱导的ROS产生和ECM表达。敲低内在XBP 1 S表达诱导ROS产生和ECM表达增加。补充过氧化氢可逆转Ad-XBP 1 S转染对ECM合成的抑制作用。P47 phox在HG处理的MC中增加。Ad-XBP 1 S转染逆转HG诱导的p47 phox表达增加，而XBP 1 S敲低上调p47 phox表达。在糖尿病大鼠肾皮质中，XBP 1 S的表达减少，而p47 phox、IV型胶原和纤维连接蛋白的表达增加。这些结果表明ER应激的XBP 1 S途径参与了HG诱导的氧化应激和ECM合成。XBP 1 S调控ROS形成的下游靶点可能是NADPH氧化酶。
Recent evidences suggest that endoplasmic reticulum (ER) stress was involved in multi pathological conditions, including diabetic nephropathy (DN). X-box binding protein 1(XBP1), as a key mediator of ER stress, has been proved having the capability of preventing oxidative stress. In this study, we investigated the effects of spliced XBP1 (XBP1S), the dominant active form of XBP1, on high glucose (HG)-induced reactive oxygen species (ROS) production and extracellular matrix (ECM) synthesis in cultured renal mesangial cells (MCs) and renal cortex of STZ-induced diabetic rats. Real time PCR and Western blot were used to evaluate the mRNA and protein levels respectively. Transfection of recombinant adenovirus vector carrying XBP1S gene (Ad-XBP1S) was used to upregulate XBP1S expression. XBP1S siRNA was used to knockdown XBP1S expression. ROS level was detected by dihydroethidium (DHE) fluorescent probe assay. The results showed that HG treatment significantly reduced XBP1S protein and mRNA level in the cultured MCs while no obvious change was observed in unspliced XBP1 (XBP1U). In the mean time, the ROS production, collagen IV and fibronectin expressions were increased. Diphenylene-chloride iodonium (DPI), a NADPH oxidase inhibtor, prevented HG-induced increases in ROS as well as collagen IV and fibronectin expressions. Transfection of Ad-XBP1S reversed HG-induced ROS production and ECM expressions. Knockdown intrinsic XBP1S expression induced increases in ROS production and ECM expressions. Supplementation of supreoxide reversed the inhibitory effect of Ad-XBP1S transfection on ECM synthesis. P47phox was increased in HG-treated MCs. Ad-XBP1S transfection reversed HG-induced p47phox increase while XBP1S knockdown upregulated p47phox expression. In the renal cortex of diabetic rats, the expression of XBP1S was reduced while p47phox, collagen IV and fibronectin expression were elevated. These results suggested that XBP1S pathway of ER stress was involved in HG-induced oxidative stress and ECM synthesis. A downstream target of XBP1S in regulating ROS formation might be NADPH oxidase.
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影响因子：
11.4
作者：
Lee, AH;Chu, GC;Glimcher, LH
通讯作者：
Glimcher, LH
影响因子：
3.5
作者：
Geoffroy, K;Troncy, L;El Bawab, S
通讯作者：
El Bawab, S
DOI：
10.1152/ajprenal.00248.2009
发表时间：
2010-01-01
影响因子：
4.2
作者：
Jaimes, Edgar A.;Hua, Ping;Raij, Leopoldo
通讯作者：
Raij, Leopoldo
影响因子：
5.3
作者：
Chen, Jianchun;Chen, Jian-Kang;Harris, Raymond C.
通讯作者：
Harris, Raymond C.
影响因子：
14.9
作者：
Shinya S;Kadokura H;Imagawa Y;Inoue M;Yanagitani K;Kohno K
通讯作者：
Kohno K