Down‐regulated long non‐coding RNA ANRIL restores the learning and memory abilities and rescues hippocampal pyramidal neurons from apoptosis in streptozotocin‐induced diabetic rats via the NF‐κB signaling pathway
Authors: Yong-Jian Wang
Dong-Mei Wu
Xin-Rui Han
Bin Hu
Chun-Hui Sun
Xin Wen
Min Shen
Liang Wang
Meng-Qiu Li
Jun Lu
Shan Wang
Shao-Hua Fan
Qun Shan
Zi-Feng Zhang
Yuan-Lin Zheng
Journal: Journal of Cellular Biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: antisense non-coding RNA in the INK4 locus; diabetes; hippocampal neuronal apoptosis; learning and memory abilities; nuclear factor kappa B
Affiliations: Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu ProvinceSchool of Life ScienceJiangsu Normal UniversityXuzhouJiangsuChina, College of Health SciencesJiangsu Normal UniversityXuzhouJiangsuChina

Abstract:  Diabetes often causes learning and memory deficits, which leads to unfavorable behavioral performance. In this study, we investigated the effects of long non-coding RNA (lncRNA) ANRIL on learning, memory abilities, and hippocampal neuronal apoptosis via the NF-κB signaling pathway in streptozotocin (STZ)-induced diabetic rats. After successful establishment of diabetic rat models, the subjects were then assigned into the DM, DM + si-ANRIL, DM + si-negative control (si-NC) groups, as well as an additional normal group. Morris water maze test was employed to assess behavioral performance of rats, followed by the recording of body weight and blood glucose levels. Expressions of ANRIL, NF-κB signaling pathway-related, and apoptosis-related genes were examined by qRT-PCR and western blotting. Rat hippocampus expression levels of cleaved-caspase-3 were determined by immunofluorescence. Cell apoptosis was examined by TUNEL assay. Versus to the normal group, revealed there to be activation of the NF-κB signaling pathway, decreased weight, increased blood glucose, increased escape latency, reduced residence time, memory impairment, increased cleaved-caspase-3 expression, and increased apoptosis were detected in the DM and DM + si-NC groups. The DM + si-ANRIL group exhibited inhibited NF-κB signaling pathway, weight loss, decreased blood glucose, recovered memory, decreased cleaved-caspase-3 expression and reduced apoptosis compared to the DM group, with higher weight of rats, lower blood glucose levels, and stronger memory abilities in the DM + si-ANRIL group. Taken together, these findings indicate that silencing lncRNA ANRIL promotes memory recovery and decreases hippocampal neurons apoptosis in diabetic rats through the inhibition of the NF-κB signaling pathway. 

Retracted: Yes

PubPeer comments:
  FIGURE 5 Expressions of NF-u03baB signaling pathway-related proteins in rat hippocampus by Western blotting among the normal group, DM, DM + si-ANRIL, DM + si-NC, and DM + si-ANRIL groups. Note: A, the protein bands of NF-u03baB signaling pathway-related proteins following Western blotting .  FIGURE 6 Expression of apoptosis-related proteins in rat hippocampus by Western blotting among the normal group, the DM group, the DM + si-ANRIL group, and the DM + si-NC group. Note: A, the protein bands of apoptosis-related proteins following Western blotting . ![file] The protein blobs seem to come from the same restricted gamut shown in this montage. ![] Clockwise from upper left, Fig 6B,  protein band graph , from  MicroRNA-374 Exerts Protective Effects by Inhibiting SP1 Through Activating the PI3K Akt Pathway in Rat Models of Myocardial Ischemia-Reperfusion After Sevoflurane Preconditioning [(Zhang et al 2018)] . Fig 4D,  GLP-1R and CREB proteins in the myocardium of mice in each group examined by western blotting , from  Suppression of Long Non-Coding RNA LINC00652 Restores Sevoflurane-Induced Cardioprotection Against Myocardial Ischemia-Reperfusion Injury by Targeting GLP-1R Through the cAMP PKA Pathway in Mice [(Zhang et al 2018)] . Fig. 6B,  ELISA and western blot analysis were applied to determine the miR- 495 expression and mRNA and protein expression of NLRP3, TNF-u03b1, IL-1u03b2, IL-18 and caspase-1 , from  MicroRNA-495 Ameliorates Cardiac Microvascular Endothelial Cell Injury and Inflammatory Reaction by Suppressing the NLRP3 Inflammasome Signaling Pathway [(Zhou et al 2018)] . Fig 6B,  WB images showing protein expression of different genes in each group , from  MicroRNA-27a Suppresses Detrusor Fibrosis in Streptozotocin-Induced Diabetic Rats by Targeting PRKAA2 Through the TGF-u03b21 Smad3 Signaling Pathway . [(Wu et al 2018)] . Fig 11E,  protein bands of each factor in ASMCs of asthma mice in each group detected by western blot analysis , from  MicroRNA-485 Modulates the TGF-u03b2 Smads Signaling Pathway in Chronic Asthmatic Mice by Targeting Smurf2 . [(Wang et al 2018)] . , 
 Comparing Fig 6A *[right]* with *[left]* 9C from [Han, Ju Geng (2018)] . The resemblance between a Bcl-2 band and a GAPDH loading control is what caught my attention. ![file] UPDATE - I have added Fig 3E from [Wen et al (2018)] *[below]* ![file] , 
 Fig 9.  Ultrastructure of pyramidal neurons in the hippocampal CA1 region observed under the transmission electron microscope among the normal group, DM, DM + si-ANRIL, DM + si-NC groups . ![file] Or possibly rat corpus striatum: Fig. 5 from [Wu et al (2018)] .  Inhibition of miR-200a or inactivation of cAMP improves the striatum structure in PD rats as detected by electron microscopy . ![] Or possibly breast-carcinoma cell lines Fig 3D from [Gu et al (2018)] :  GAS5 inhibits autophagy in breast cancer cells. ... (D) Transmission electron microscopy was used to detect autophagy in MDA-MB-23l and MCF-7 cells after overexpression or down-regulation of GAS5 . ![file] Or possibly hippocampus again. Fig 1 from [Liu et al (2018)] : ![] Fig. 2 from [Yu et al (2018)] . LFA-1 gene silencing improves ultrastructure of hippocampal neurons in CA1 area. ![] , 
 Retracted [19 July 2021] . ---------------------------------------------- The above article, published online on 30 March 2018 in Wiley Online Library has been retracted by agreement between the journal s Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. The retraction has been agreed following an investigation based on allegations raised by a third party. A detailed investigation revealed that several image elements of the experimental data were published elsewhere in a different scientific context. Thus, the editors consider the conclusions of this article to be invalid. , 
 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 31:  Xin Wen # , Xin-Rui Han # , Yong-Jian Wang # , Shan Wang, Min Shen, Zi-Feng Zhang, Shao-Hua Fan, Qun Shan, Liang Wang, Meng-Qiu Li, Bin Hu, Chun-Hui Sun, Dong-Mei Wu*, Jun Lu*, and Yuan-Lin Zheng*. **Down-regulated long non-coding RNA ANRIL restores the learning and memory abilities and rescues hippocampal pyramidal neurons from apoptosis in streptozotocin-induced diabetic rats via the NF-u03baB signaling**. Journal of Cellular Biochemistry, 2018, 119(7):5821-5833.u201d (with fund numbers 81271225, 81171012, 30950031, 81571055, 81400902, 81570531, 81672731) .... 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 ,