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GSK3β inhibits HDAC4 to Alleviate Intravertebral Disc Degeneration Through KLF5 and ASK1 Downregulation
29 Pages Posted: 11 Aug 2020
More...Abstract
Intervertebral disc degeneration (IDD) is a major cause of lower back pain, yet the molecular mechanisms are not fully understood. Therefore, the effects of HDAC4 and its upstream and downstream signaling molecules on mediating IDD are determined in this study. IDD mouse model was established by injury of the vertebrate. Nucleus pulposus (NP) cells were collected for culture and transfection. Disc height index (DHI) was determined by X-ray. Pain in IDD mice was evaluated by mechanical and thermal sensitivity testes. Interaction between GSK3β and HDAC4 was assessed by co-immunoprecipitation. The effect of HDAC4 on KLF5 acetylation was determined by immunoprecipitation. Promoter region binding was assessed by chromatin immunoprecipitation. NP cells apoptosis was determined by TUNEL assay. Degradation of extracellular matrix was studied by safranin O fast green staining. Morphology of intervertebral disc tissue was determined by HE staining. mRNA and protein expression of GSK3β, HDAC4, phosphorylated HDAC4, KLF5, and ASK1 were determined by qRT-PCR and Western blotting, respectively. HDAC4 was upregulated while GSK3β was downregulated in IDD mice. HDAC4 silencing alleviated IDD. GSK3β promoted HDAC4 phosphorylation and degradation. HDAC4 promoted ASK1 expressed through, leading to enhanced IDD. GSK3β overexpression increased DHI, inhibited NP cell apoptosis, alleviated disc degeneration, and increased mechanical and thermal pain thresholds in IDD mice. The addition of HDAC4 overexpression reversed these effects of GSK3β. In conclusion, HDAC4 was degraded by GSK3β, leading to reduced IDD through KLF5 and ASK1 downregulation.
Funding Statement: None.
Declaration of Interests: The authors declare no conflicts of interest.
Ethics Approval Statement: The above experiments were approved by the Animal Ethics Committee in Guangxi University of Chinese Medicine, Guangxi International Zhuang Medicine Hospital.
Keywords: intravertebral disc degeneration; GSK3β; KLF5; ASK1; HDAC4
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