Augmenter of Liver Regeneration Alleviates Renal Hypoxia-Reoxygenation Injury by Regulating Mitochondrial Dynamics in Renal Tubular Epithelial Cells

Long, Rui-ting,Peng, Jun-bo,Huang, Li-li,Jiang, Gui-ping,Liao, Yue-juan,Sun, Hang,Hu, Yu-dong,Liao, Xiao-hui Korean Society for Molecular and Cellular Biology 2019 Molecules and cells Vol.42 No.12

Mitochondria are highly dynamic organelles that constantly undergo fission and fusion processes that closely related to their function. Disruption of mitochondrial dynamics has been demonstrated in acute kidney injury (AKI), which could eventually result in cell injury and death. Previously, we reported that augmenter of liver regeneration (ALR) alleviates renal tubular epithelial cell injury. Here, we gained further insights into whether the renoprotective roles of ALR are associated with mitochondrial dynamics. Changes in mitochondrial dynamics were examined in experimental models of renal ischemia-reperfusion (IR). In a model of hypoxia-reoxygenation (HR) injury in vitro, dynamin-related protein 1 (Drp1) and mitochondrial fission process protein 1 (MTFP1), two key proteins of mitochondrial fission, were downregulated in the Lv-ALR + HR group. ALR overexpression additionally had an impact on phosphorylation of Drp1 Ser637 during AKI. The inner membrane fusion protein, Optic Atrophy 1 (OPA1), was significantly increased whereas levels of outer membrane fusion proteins Mitofusin-1 and -2 (Mfn1, Mfn2) were not affected in the Lv-ALR + HR group, compared with the control group. Furthermore, the mTOR/4E-BP1 signaling pathway was highly activated in the Lv-ALR + HR group. ALR overexpression led to suppression of HR-induced apoptosis. Our collective findings indicate that ALR gene transfection alleviates mitochondrial injury, possibly through inhibiting fission and promoting fusion of the mitochondrial inner membrane, both of which contribute to reduction of HK-2 cell apoptosis. Additionally, fission processes are potentially mediated by promoting tubular cell survival through activating the mTOR/4E-BP1 signaling pathway.

Transposon <i>Ac/Ds</i> -induced chromosomal rearrangements at the rice <i>OsRLG5</i> locus

Xuan, Yuan Hu,Piao, Hai Long,Je, Byoung Il,Park, Soon Ju,Park, Su Hyun,Huang, Jin,Zhang, Jian Bo,Peterson, Thomas,Han, Chang-deok Oxford University Press 2011 Nucleic acids research Vol.39 No.22

<P>Previous studies have shown that pairs of closely-linked <I>Ac/Ds</I> transposable elements can induce various chromosomal rearrangements in plant genomes. To study chromosomal rearrangements in rice, we isolated a line (<I>OsRLG5-161)</I> that contains two inversely-oriented <I>Ds</I> insertions in <I>OsRLG5</I> (<I>Oryza sativa</I> Receptor like kinase Gene 5). Among approximately 300 plants regenerated from <I>OsRLG5-161</I> heterozygous seeds, 107 contained rearrangements including deletions, duplications and inversions of various sizes. Most rearrangements were induced by previously identified alternative transposition mechanism. Furthermore, we also detected a new class of rearrangements that contain juxtaposed inversions and deletions on the same chromosome. We propose that these novel alleles were generated by a previously unreported type of alternative transposition reactions involving the 5′ and 3′ termini of two inversely-oriented <I>Ds</I> elements located on the same chromatid. Finally, 11% of rearrangements contained inversions resulting from homologous recombination between the two inverted <I>Ds</I> elements in <I>OsRLG5-161</I>. The high frequency inheritance and great variety of rearrangements obtained suggests that the rice regeneration system results in a burst of transposition activity and a relaxation of the controls which normally limit the transposition competence of individual <I>Ds</I> termini. Together, these results demonstrate a greatly enlarged potential of the <I>Ac/Ds</I> system for plant chromosome engineering.</P>

Production of a Phage-displayed Mouse ScFv Antibody against Fumonisin B1 and Molecular Docking Analysis of Their Interactions

Zu-Quan Hu,He-Ping Li,Jin-Long Liu,Sheng Xue,An-Dong Gong,Jing-Bo Zhang,Yu-Cai Liao 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

Fumonisins produced by Fusarium pathogens are mycotoxins present in maize and other grains in the field as well as during storage worldwide and pose a serious threat to humans and domestic animals. Fumonisin B consists of different chemotypes, and fumonisin B1 (FB1) is the most predominant fumonisin found in food/feed commodities. Recombinant antibody can be deployed to analyze the fumonisin toxicological mechanism and develop a simple and cost-effective method for the detection of fumonisins, which is vitally important for monitoring and preventing fumonisins from entering food/feed chains. In this study, FB1 conjugated to keyhole limpet hemocyanin was used to immunize mice, from which RNA was isolated to construct a recombinant antibody library. Successive panning of the library by phage display was used to select monoclonal phage clones reactive to FB1 conjugated to bovine serum albumin. Subsequent phage ELISA and sequencing analyses revealed four different reactive scFv antibodies specific to FB1. Soluble expression and ELISA analysis showed that one scFv antibody, FBMA1, had the highest reactivity and could be purified from bacterial cells in large quantities. Surface plasmon resonance measurements further revealed that the FBMA1 scFv antibody had a binding kinetics of KD = 1.89 × 10–7 M. Molecular modeling and docking analyses suggested that the FBMA1 antibody shaped a proper cavity to embed the whole FB1 molecule and that a steady-state complex was formed relying on intermolecular forces, including hydrogen bonding, electrostatic force and hydrophobic interactions. Thus, the scFv antibody can be applied for mechanistic studies of intermolecular interactions and fumonisin toxicity, and for the development of an immunoassay for fumonisin-contaminated food/feed samples.

Selective miRNA Expression Profile in Chronic Myeloid Leukemia K562 Cell-derived Exosomes

Feng, Dan-Qin,Huang, Bo,Li, Jing,Liu, Jing,Chen, Xi-Min,Xu, Yan-Mei,Chen, Xin,Zhang, Hai-Bin,Hu, Long-Hua,Wang, Xiao-Zhong Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.12

Background: Chronic myeloid leukemia (CML) is a myeloproliferative disorder of hematopoietic stem cell scarrying the Philadelphia (Ph) chromosome and an oncogenic BCR-ABL1 fusion gene. The tyrosine kinase inhibitor (TKI) of BCR-ABL1 kinase is a treatment of choice for control of CML. Objective: Recent studies have demonstrated that miRNAs within exosomes from cancer cells play crucial roles in initiation and progression. This study was performed to assess miRNAs within exosomes of K562 cells. Methods: miRNA microarray analysis of K562 cells and K562 cell-derived exosomes was conducted with the 6th generation miRCURYTM LNA Array (v.16.0). Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were also carried out. GO terms and signaling pathways were categorized into 66 classes (including homophilic cell adhesion, negative regulation of apoptotic process, cell adhesion) and 26 signaling pathways (such as Wnt). Results: In exosomes, 49 miRNAs were up regulated as compared to K562 cells, and two of them were further confirmed by quantitative real-time PCR. There are differentially expressed miRNAs between K562 cell derived-exosomes and K562 cells. Conclusion: Selectively expressed miRNAs in exosomes may promote the development of CML via effects on interactions (e.g. adhesion) of CML cells with their microenvironment.

Nonselective Blocking of the Sympathetic Nervous System Decreases Detrusor Overactivity in Spontaneously Hypertensive Rats

Kim, Khae-Hawn,Jin, Long-Hu,Choo, Gwoan-Youb,Lee, Hun-Jae,Choi, Bo-Hwa,Kwak, Jiyeon,Yoon, Sang-Min,Park, Chang-Shin,Lee, Tack Molecular Diversity Preservation International (MD 2012 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.13 No.4

<P>The involuntary dual control systems of the autonomic nervous system (ANS) in the bladder of awake spontaneously hypertensive rats (SHRs) were investigated through simultaneous registrations of intravesical and intraabdominal pressures to observe detrusor overactivity (DO) objectively as a core symptom of an overactive bladder. SHRs (<I>n</I> = 6) showed the features of overactive bladder syndrome during urodynamic study, especially DO during the filling phase. After injection of the nonselective sympathetic blocking agent labetalol, DO disappeared in 3 of 6 SHRs (50%). DO frequency decreased from 0.98 ± 0.22 min<SUP>−1</SUP> to 0.28 ± 0.19 min<SUP>−1</SUP> (<I>p</I> < 0.01), and DO pressure decreased from 3.82 ± 0.57 cm H<SUB>2</SUB>O to 1.90 ± 0.86 cm H<SUB>2</SUB>O (<I>p</I> < 0.05). This suggests that the DO originating from the overactive parasympathetic nervous system is attenuated by the nonselective blocking of the sympathetic nervous system. The detailed mechanism behind this result is still not known, but parasympathetic overactivity seems to require overactive sympathetic nervous system activity in a kind of balance between these two systems. These findings are consistent with recent clinical findings suggesting that patients with idiopathic overactive bladder may have ANS dysfunction, particularly a sympathetic dysfunction. The search for newer and better drugs than the current anticholinergic drugs as the mainstay for overactive bladder will be fueled by our research on these sympathetic mechanisms. Further studies of this principle are required.</P>

Augmenter of Liver Regeneration Alleviates Renal Hypoxia-Reoxygenation Injury by Regulating Mitochondrial Dynamics in Renal Tubular Epithelial Cells

Xiao-hui Liao,Rui-ting Long,Jun-bo Peng,Li-li Huang,Gui-ping Jiang,Yue-juan Liao,Hang Sun,Yu-dong Hu 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.12

Mitochondria are highly dynamic organelles that constantly undergo fission and fusion processes that closely related to their function. Disruption of mitochondrial dynamics has been demonstrated in acute kidney injury (AKI), which could eventually result in cell injury and death. Previously, we reported that augmenter of liver regeneration (ALR) alleviates renal tubular epithelial cell injury. Here, we gained further insights into whether the renoprotective roles of ALR are associated with mitochondrial dynamics. Changes in mitochondrial dynamics were examined in experimental models of renal ischemia-reperfusion (IR). In a model of hypoxia-reoxygenation (HR) injury in vitro, dynamin-related protein 1 (Drp1) and mitochondrial fission process protein 1 (MTFP1), two key proteins of mitochondrial fission, were downregulated in the Lv-ALR + HR group. ALR overexpression additionally had an impact on phosphorylation of Drp1 Ser637 during AKI. The inner membrane fusion protein, Optic Atrophy 1 (OPA1), was significantly increased whereas levels of outer membrane fusion proteins Mitofusin-1 and -2 (Mfn1, Mfn2) were not affected in the Lv-ALR + HR group, compared with the control group. Furthermore, the mTOR/4E-BP1 signaling pathway was highly activated in the Lv-ALR + HR group. ALR overexpression led to suppression of HR-induced apoptosis. Our collective findings indicate that ALR gene transfection alleviates mitochondrial injury, possibly through inhibiting fission and promoting fusion of the mitochondrial inner membrane, both of which contribute to reduction of HK-2 cell apoptosis. Additionally, fission processes are potentially mediated by promoting tubular cell survival through activating the mTOR/4E-BP1 signaling pathway.

Aberrant microRNAs Expression in CD133+/CD326+ Human Lung Adenocarcinoma Initiating Cells from A549

Sheng Lin,Zheng-tang Chen,Jian-guo Sun,Jing-bo Wu,Hai-xia Long,Cong-hui Zhu,Tong Xiang,Hu Ma,Zhong-quan Zhao,Quan Yao,An-mei Zhang,Bo Zhu 한국분자세포생물학회 2012 Molecules and cells Vol.33 No.3

Increasing evidence demonstrates that miRNAs are in-volved in the dysregulation of tumor initiating cells (TICs) in various tumors. Due to a lack of definitive markers, cell sorting is not an ideal separation method for lung adeno-carcinoma initiating cells. In this study, we combined pa-clitaxel with serum-free medium cultivation (inverse-induc-tion) to enrich TICs from A549 cells, marked by CD133/ CD326, defined features of stemness. We next investigated aberrant microRNAs in this subpopulation compared to normal cells with miRNA microarray and found that 50 miRNAs exhibited a greater than 2-fold change in expres-sion. As further validation, 10 miRNAs were chosen to perform quantitative RT-PCR on the A549 cell line and primary samples. The results suggest that aberrant ex-pression of miRNAs such as miR-29ab, miR-183, miR-17-5p and miR-127-3P may play an important role in regulat-ing the bio-behavior of TICs.

Vp28 of Shrimp White Spot Syndrome Virus Is Involved in the Attachment and Penetration into Shrimp Cells

( Guo Hua Yi ),( Zhi Min Wang ),( Yi Peng Qi ),( Lun Guang Yao ),( Juan Qian ),( Long Bo Hu ) 생화학분자생물학회 2004 BMB Reports Vol.37 No.6

White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within OS It However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition EI,ISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.