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Abstract
Background
The method of action of Bushen Formula (BSHXF) in the treatment of intervertebral disc degeneration (IVDD) was uncovered in this work using in vivo and in vitro tests. To clarify the mechanism of action of BSHXF, we validated the rat intervertebral disc degeneration model and the nucleus pulposus cell degeneration model.
Methods
In an in vivo model of IVDD the study explores the impact of BSHXF on mitochondrial function, pro-inflammatory cytokines, pro-apoptotic factors, and matrix metalloproteinases. Additionally, it evaluates the induced degeneration of nucleus pulposus (NP) cells in an in vitro model stimulated by interleukin-1 β (IL-1β). The study measures the effects of BSHXF on both the inflammatory response and mitochondrial function.
Results
The MRI results showed that BSHXF reduced intervertebral disc volume reduction and degradation of NP tissue. HE, SO-FG and immunofluorescence further confirmed the protective effect of BSHXF on degenerative intervertebral discs. BSHXF reduced the inflammatory levels of IL-6 IL-1β and TNF-α in degenerative intervertebral disc tissue. Meanwhile, JC-1, mPTP and ROS detection revealed that BSHXF can restore mitochondrial function by regulating the expression of antioxidant proteins, playing a protective role in NP cells. Finally, the WB results showed that BSHXF can alleviate IL-1β mediate the degeneration of NP cells. BSHXF can alleviate NP cell apoptosis by inhibiting the expression of bax, cleaved caspase-3, caspase-3, and cyt-c, and increasing the expression of Bcl-2.
Conclusion
This study reveals that BSHXF inhibits the development of inflammatory factors, which may play a significant role in intervertebral disc degeneration. This implies that BSHXF is a suitable herbal medication for future research into inflammatory cytokine treatment.
Data Sharing Statement
The datasets utilized or scrutinized during this study are accessible upon reasonable request from the corresponding author.
Ethics Approval and Consent to Participate
12-week-old male Sprague-Dawley rats were procured from Jinan Pengyue Animal Co., LTD. These animals were housed in the Laboratory Animal Center of the Affiliated Hospital of Shandong University of Chinese Medicine, ensuring a barrier environment at the SPF level. The experimental protocol involving animal research received approval from the Animal Ethics Committee of the Affiliated Hospital of Shandong University of Chinese Medicine, under the Ethics approval number: 2020-35.
Author Contributions
From designing the study to its execution, encompassing data collection and analysis, all authors have made substantial contributions to this manuscript; have been actively engaged in drafting, revising, and reviewing these provisions. Moreover, all authors have given their final approval for the forthcoming version, collectively agreeing to assume responsibility for all facets of the work.
Disclosure
The authors claim that they have no known competing financial interests or personal relationships that might affect this work.