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        Grem1 accelerates nucleus pulposus cell apoptosis and intervertebral disc degeneration by inhibiting TGF-β-mediated Smad2/3 phosphorylation

        Chen Shunlun,Lei Linchuan,Li Zemin,Chen Fan,Huang Yuming,Jiang Guowei,Guo Xingyu,Zhao Zhuoyang,Liu Hui,Wang Hua,Liu Caijun,Zheng Zhaomin,Wang Jianru 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

        Intervertebral disc degeneration (IVDD) is a main cause of low back pain, and inflammatory factors play key roles in its pathogenesis. Gremlin-1 (Grem1) was reported to induce an inflammatory response in other fields. This study aimed to investigate the mechanisms of Grem1 in the degenerative process of intervertebral discs. Dysregulated genes were determined by analyzing microarray profiles. The expression of Grem1 in 17 human disc samples (male:female = 9:8) and rat models (n = 5 each group) was measured by western blotting (WB), real-time quantitative PCR (RT-qPCR), and immunohistochemistry (IHC). The regulatory effects of Grem1 on apoptosis were examined using siRNAs, flow cytometry, immunofluorescence (IF), and WB. The therapeutic effect was evaluated by locally injecting specific Grem1 siRNA into IVDD rats. The expression of Grem1 was significantly increased in human degenerative intervertebral discs; furthermore, the expression of Grem1 positively correlated with the level of intervertebral disc degeneration. Grem1 was significantly overexpressed in tumor necrosis factor (TNF)-α-induced degenerative NP cells. Apoptosis in degenerative NP cells transfected with siRNA targeting Grem1 was significantly lower than that in the control group. Specific Grem1 siRNA markedly repressed the development of IVDD in surgery-induced IVDD rats. These results indicated that the expression of Grem1 was positively correlated with the severity of intervertebral disc degeneration, and Grem1 siRNA could inhibit Grem1-induced apoptosis and extracellular matrix alterations by mediating the TGF-β/Smad signaling pathway. This study may provide a therapeutic strategy for alleviating inflammation-induced apoptosis associated with intervertebral disc degeneration.

      • SCIESCOPUSKCI등재

        Effect of additives and filling methods on whole plant corn silage quality, fermentation characteristics and in situ digestibility

        Jiao, Ting,Lei, Zhaomin,Wu, Jianping,Li, Fei,Casper, David P.,Wang, Jianfu,Jiao, Jianxin Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.11

        Objective: This project aimed to evaluate the effects of both different additives and filling methods on nutritive quality, fermentation profile, and in situ digestibility of whole plant corn silage. Methods: Whole plant corn forage harvested at 26.72% dry matter (DM) was chopped and treated with two filling methods, i) fill silos at one time (F1), ii) fill silos at three times (F3), packing samples into one/three silo capacity at the first day, another one/three capacity at the second day, then one/three at the third day, three replicates. For each replicate, samples were treated with three additives, i) control (CTRL, no additive), ii) Sila-Max (MAX, Ralco Nutrition Inc., Marshall, MN, USA), and iii) Sila-Mix (MIX, Ralco Nutrition Inc., USA). With three replicates of each secondary treatment, there were nine silos, 54 silos in total. Each silo had a packing density of 137.61 kg of DM/m3. All silos were weighed and stored in lab at ambient temperature. Results: After 60 d of ensiling, all items showed good silage fermentation under MAX filled one time or three times (p<0.01). Higher silage quality for all additives was obtained at filling one time than that filled three times (p<0.01). The highest DM and lowest DM loss rate (DMLR) occurred to MAX treatment at two filling methods (p<0.01); Digestibility of acid detergent fiber, neutral detergent fiber (NDF), and curde protein had the same results as silage quality (p<0.01). Yield of digestible DM and digestible NDF also showed higher value under MAX especially for filling one time (p<0.05). Conclusion: All corn silages showed good fermentation attributes (pH<4.0). The forage filled one time had higher silage quality than that filled three times (p<0.01). MAX with homofermentative lactic acid bacteria enhanced the lactic acid fermentation, silage quality and nutrient digestibility, and so improved the digestible nutrient yield.

      • KCI등재

        Exploring effects of different male parent crossings on sheep muscles and related regulatory genes using mRNA-Seq

        Shi Jinping,Zhang Quanwei,Song Yali,Lei Zhaomin,Fu Lingjuan,Cheng Shuru 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.8

        Objective: With improvements in living standards and increase in global population, the demand for meat products has been increasing; improved meat production from livestock could effectively meet this demand. In this study, we examined the differences in the muscle traits of different male crossbred sheep and attempted to identify key genes that regulate these traits. Methods: Dubo sheep×small-tailed Han sheep (DP×STH) and Suffolk×small-tailed Han sheep (SFK×STH) were selected to determine meat quality and production performance by Masson staining. Transcriptome sequencing and bioinformatic analysis were performed to identify differentially expressed genes (DEGs) related to meat quality. The presence of DEGs was confirmed by real-time polymerase chain reaction. Results: The production performance of SFK×STH sheep was better than that of DP×STH sheep, but the meat quality of DP×STH sheep was better than that of SFK×STH sheep. The muscle fiber diameter of DP×STH sheep was smaller than that of SFK×STH sheep. Twentytwo DEGs were identified. Among them, four gene ontology terms were related to muscle traits, and three DEGs were related to muscle or muscle fibers. There were no significant differences in the number of single nucleotide mutations and mutation sites in the different male parent cross combinations. Conclusion: This study provides genetic resources for future sheep muscle development and cross-breeding research. Objective: With improvements in living standards and increase in global population, the demand for meat products has been increasing; improved meat production from livestock could effectively meet this demand. In this study, we examined the differences in the muscle traits of different male crossbred sheep and attempted to identify key genes that regulate these traits.Methods: Dubo sheep×small-tailed Han sheep (DP×STH) and Suffolk×small-tailed Han sheep (SFK×STH) were selected to determine meat quality and production performance by Masson staining. Transcriptome sequencing and bioinformatic analysis were performed to identify differentially expressed genes (DEGs) related to meat quality. The presence of DEGs was confirmed by real-time polymerase chain reaction.Results: The production performance of SFK×STH sheep was better than that of DP×STH sheep, but the meat quality of DP×STH sheep was better than that of SFK×STH sheep. The muscle fiber diameter of DP×STH sheep was smaller than that of SFK×STH sheep. Twenty-two DEGs were identified. Among them, four gene ontology terms were related to muscle traits, and three DEGs were related to muscle or muscle fibers. There were no significant differences in the number of single nucleotide mutations and mutation sites in the different male parent cross combinations.Conclusion: This study provides genetic resources for future sheep muscle development and cross-breeding research.

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