Repression of microRNA-382 inhibits glomerular mesangial cell proliferation and extracellular matrix accumulation via FoxO1 in mice with diabetic nephropathy
Authors: Yong-Jian Wang
Dong-Mei Wu
Xin-Rui Han
Bin Hu
Chun-Hui Sun
Juan Zhuang
Xin Wen
Min Shen
Meng-Qiu Li
Jun Lu
Shan Wang
Shao-Hua Fan
Qun Shan
Zi-Feng Zhang
Yuan-Lin Zheng
Journal: Cell Proliferation (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: FoxO1; diabetic nephropathy; extracellular matrix; glomerular mesangial cells; microRNA-382
Affiliations: Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, China., College of Health Sciences, Jiangsu Normal University, Xuzhou, China., School of Environment Science and Spatial Informatics, China University of Mining and Technology, Xuzhou, China., Jiangsu Key Laboratory for Eco-Agricultural Biotechnology around Hongze Lake, School of Life Sciences, Huaiyin Normal University, Huaian, China.

Abstract:  Objectives: Diabetic nephropathy (DN) is a nerve damaging disorder, characterized by glomerular mesangial cell expansion and accumulation of extracellular matrix (ECM) proteins. In this study, we aimed to investigate mesangial cell proliferation and ECM accumulation when promoting or suppressing endogenous miR-382 in glomerular mesangial cells of DN. Materials and methods: Model establishment consisted of DN induction by streptozotocin (STZ) in mice. The underlying regulatory mechanisms of miR-382 were analysed in concert with the treatment of miR-382 mimics, miR-382 inhibitors or siRNA against FoxO1 in cultured glomerular mesangial cells isolated from DN mice. Results: FoxO1 was identified as the downregulated gene in DN based on the microarray data of GSE1009. We found that miR-382 was significantly upregulated in renal tissues of DN mice and its downregulation dephosphorylated FoxO1, reduced glomerular mesangial cell proliferation and ECM accumulation in vitro. The determination of luciferase activity suggested that miR-382 negatively targeted FoxO1. Expectedly, distinct levels of phosphorylated FoxO1 were observed in the renal cortices of DN mice, while the silencing of FoxO1 was found to increase glomerular mesangial cell proliferation and ECM accumulation in vitro. Reduced glomerular mesangial cell proliferation and ECM accumulation elicited by miR-382 inhibitors were reversed by silencing FoxO1. Conclusions: This study demonstrates miR-382 suppression exerts a potent anti-proliferative effect that may be applied to inhibit glomerular mesangial cell proliferation and ECM accumulation in DN. 

Retracted: No

PubPeer comments:
 *[Left]* FIGURE 5B.  Upregulation of miR-382 negatively regulates FoxO1 and increases ECM accumulation in the glomerular mesangial cells.... Panels B and C, western blot analysis was used to determine the protein expressions of FoxO1, p-FoxO1, FN, Col I, Col IV and PAI-1; the protein expression of FoxO1 does not differ among the six groups . *[Right]* Figure 6A,C from [Wen et al (2018)] .  Expressions of EMT-related proteins (E-cadherin and Vimentin) detected by western blotting Note: A and C, expressions of EMT-related proteins (E-cadherin and Vimentin) in A549 and H1975 cells co-transfected with si-SPRY4-IT1 and siEZH2 . A loading control seems to be used in both. ![file] , 
 #1 It is also worth comparing Fig 5B with 3B,  When the mice were induced by STZ, miR-382 is upregulated while the FoxO1 is reciprocal, and extracellular matrix proteins are increased in renal cortices of DN mice. ... Panels B and C, western blot analysis was used to determine the protein expressions of FoxO1, p-FoxO1, FN, Col I, Col IV and PAI-1; the protein expression of FoxO1 does not differ after STZ inducement. ![file] Another sighting of the control band: Fig 5B from [Wu et al (2018)] .  Expression of Wnt u03b2-catenin signaling pathway-related factors in rats of each group. ... B, relative protein expression of u03b2-catenin, GSK-3u03b2, GSK-3u03b2-Ser9 and cyclinD1 detected by western blot . ![] , 
 Fig 7C.  Upregulation of miR-382 or glomerular FoxO1 affects cell cycle distribution while inhibiting apoptosis in glomerular mesangial cells... Panel C, glomerular mesangial cells in the scatter plots in which the upper left quadrant identifies the necrotic cells (annexin V- PI+), the upper right quadrant identifies the late apoptotic cells (annexin V+ PI+), the lower left quadrant identifies the live cells (annexin V- PI-) and the lower right quadrant identifies the early apoptotic cells (annexin V+ PI-). ![] Fig 6A,C from [Wen et al (2017)] : ![file] Fig. 10A from [Sun et al (2018)] .  Cell apoptosis after transfection in the control, blank, NC, miR-592-5p mimics, miR-592-5p inhibitors, siRNA-PTGDR, and miR-592-5p inhibitors + siRNA-PTGDR groups. Note: A, cell apoptosis diagram for each group . ![file] Boxed plots are obviously not identical, but have more cells in common than one would expect from unrelated experiments involving completely unrelated cell-lines and conditions. Most apparent in the upper left quadrants. For instance, ![] ![file] UPDATE: Fig 10A of [Wang et al (2018)] : ![] , 
 Assessed here, as part of the first of 10 broad investigations: https: www.nsfc.gov.cn publish portal0 tab442 info85495.htm **Decisions on handling scientific research misconduct cases investigated and dealt with in 2022 (first batch)** Date 2022-05-13 Source: u3000 Author: u3000 u3010 Large, Middle and Small u3011 u3010Printu3011 u3010Closeu3011 u3000u3000The National Natural Science Foundation of China resolutely implements the spirit of the  Several Opinions on Further Strengthening the Construction of Scientific Research Integrity and  Opinions on Further Promoting the Spirit of Scientists and Strengthening the Construction of Style and Study Style by the General Office of the Central Committee of the Communist Party of China and the General Office of the State Council. Horizontal science and technology are self-reliant and self-reliant, continue to carry out scientific research integrity construction, and seriously investigate and deal with scientific research misconduct involving the National Natural Science Foundation of China. The Supervision Committee of the National Natural Science Foundation of China conducts investigations, verifications and comprehensive judgments on the accepted scientific research integrity cases, in accordance with the Regulations of the National Natural Science Foundation of ) and  Measures for Investigation and Handling of Scientific Research Misconduct in National Natural Science Foundation of China and other regulations and systems to propose handling suggestions. After deliberation at the second executive meeting of the National Natural Science Foundation of China in 2022, it was decided to deal with the relevant parties involved. The 10 handling decisions (online release version) that have been criticized and criticized are hereby announced. -------------------------------- About the existence of papers published on Wu Dongmei, Zheng Yuanlin, Lu Jun, etc. - Falsification of pictures or data, fabrication of research process, use of others signatures without consent, and false information in project final reports - processing decision - National Science and Technology Financial Supervisory Office [2022] No. 1 u3000u3000The Supervision Committee of the National Natural Science Foundation of China launched an investigation into several papers of Jiangsu Normal University, including Wu Dongmei, Zheng Yuanlin, and Jun Jun, for suspected academic misconduct. The papers involved are as follows: ... Paper 16:  Shan Wang # , Xin Wen # , Xin-Rui Han, Yong-Jian Wang, Min Shen, Shao-Hua Fan, Juan Zhuang, Zi-Feng Zhang, Qun Shan,Meng-Qiu Li, Bin Hu, Chun -Hui Sun, Dong-Mei Wu*, Jun Lu*, Yuan-Lin Zheng*. **Repression of microRNA-382 inhibits glomerular mesangial cell proliferation and extracellular matrix accumulation via FoxO1 in mice with diabetic nephropathy**. Cell Proliferation, 2018, e12462.u201d (marked with fund numbers 81271225, 31201039, 81171012, 30950031, 81571055, 81400902) ... u3000u3000According to the investigation of the supporting unit, the above 35 papers involved all have the problem of improper use of pictures or data fraud, and the pictures or data in question are all provided by third-party companies. Among the 35 papers involved, 34 (except paper 21) still have the problem of using others signatures without consent, and papers 4, 5, 9, 12, 30, and 34 also have the problem of fabricating the research process. Wu Dongmei is the author of the above 35 papers involved, of which 12 papers (papers 2, 8, 9, 10, 11, 12, 14, 15, 25, 26, 30, 35) are the first author and 22 papers (Paper 1, 4, 5, 6, 7, 13, 16, 17, 18, 19, 20, 21, 22, 23, 24, 27, 28, 29, 31, 32, 33, 34) Corresponding author, They should be primarily responsible for the objective results of falsifying pictures or data, using others signatures without consent, and fabricating research processes; Zheng Yuanlin and Lu Jun, as the co-corresponding authors of the 35 papers involved, were negligent in reviewing and took secondary responsibility. In addition, Wu Dongmei included 4 papers involved (papers 21, 24, 28, 29) in the final report of the National Natural Science Foundation of China (Grant No. 81400902), and Zheng Yuanlin included 9 papers involved (papers 6, 8, 10, 13, 21, 24, 26, 28, 29) included in the final report of the National Natural Science Foundation of China (Grant No. 81571055), and should be responsible for the objective results of false information in the final report. u3000u3000After deliberation at the 12th meeting of the Fifth Supervision Committee of the National Natural Science Foundation of China (Life Medicine Professional Committee) and the approval of the second executive meeting of the National Natural Science Foundation of China in 2022, it was decided to follow the thirty-fifth National Natural Science Foundation of China Regulations. Article 4, Article 17, Items 3, 4, and 9 of  Measures for Handling Misconduct in Science Fund Work by the Supervision Committee of the National Natural Science Foundation of China (for Trial Implementation) , revocation of Wu Dongmei s National Natural Science Foundation of Molecular mechanism of TRPV4-mediated domoic acid treatment-induced cognitive deficits in mice (Grant No. 81400902), recovering the allocated funds, permanently disqualifying Wu Dongmei from the National Natural Science Foundation of China (since January 18, 2022), Give Wu Dongmei a circular of criticism; decide to follow Article 35, item 4 of the  National Natural Science Foundation of China Regulations , and refer to  The National Natural Science Foundation of China Supervision Committee s Handling of Misconduct in Science Fund Work (Trial) Tenth Items 3 and 4 of seven articles, the Zheng Yuanlin National Natural Science Foundation of China project  The Mechanism of HDAC6 Activating HMGB1 RAGE Signaling Axis to Mediate Cognitive Defects in Type 2 Diabetic Mice (Approval No. 81571055), recover the allocated funds, cancel Zheng Yuanlin will be eligible to apply for the National Natural Science Foundation of China for 5 years (January 18, 2022 to January 17, 2027), and will give Zheng Yuanlin a notice of criticism; it is decided to refer to the  National Natural Science Foundation of China Supervision Committee on misconduct in the work of the National Natural Science Foundation of China. Items 3 and 4 of Article 17 of the Measures (for Trial Implementation), the Army s National Natural Science Foundation of China project application qualification will be cancelled for 5 years (January 18, 2022 to January 17, 2027), and the Army will be criticized in a circular. - National Natural Science Foundation of China - March 30, 2022 ,