Target NF-κB p65 for preventing posttraumatic joint contracture in rats

Lingpeng Kong; Yuqing Liang; Jing Hou; Weiying Zhang; Shichao Jiang

doi:10.1002/jor.25877

Target NF-κB p65 for preventing posttraumatic joint contracture in rats

J Orthop Res. 2024 May 15. doi: 10.1002/jor.25877. Online ahead of print.

Authors

Lingpeng Kong¹, Yuqing Liang², Jing Hou², Weiying Zhang³, Shichao Jiang²

Affiliations

¹ Department of Orthopedics, Second Affiliated Hospital of Soochow University, Suzhou, China.
² Department of Orthopedics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.
³ Health Management Center, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.

PMID: 38751161
DOI: 10.1002/jor.25877

Abstract

RelA/p65 is as a crucial component of the nuclear factor κB (NF-κB) signaling pathway that has a significant impact on various fibrotic diseases. However, its role in the fibrosis of tissues surrounding the joint after traumatic injury remains unclear. In this study, rats were divided into three groups: non-operated control (NC) group, p65-siRNA treated (siRNA-p65) group, and negative siRNA treated (siRNA-neg) group. Then, 10 μL (10 nmol) of p65-siRNA was injected into the joint of the siRNA-p65 group. Meanwhile, 10 μL of negative siRNA was administered to the knee joint of the operated siRNA-neg group for comparison. The rats in the NC group did not receive surgery or drug intervention. After 4 weeks of right knee fixation in each group, X-ray measurements revealed significantly reduced degree of knee flexion contracture following p65-siRNA treatment (siRNA-neg: 77.73° ± 2.799°; siRNA-p65: 105.7° ± 2.629°, p < 0.0001). Histopathological examination revealed that the number of dense fibrous connective tissues decreased following p65-siRNA inhibition. Western blot analysis revealed significantly different expression levels of fibrosis-related proteins between the siRNA-p65 and siRNA-neg groups. Immunohistochemical analysis revealed a reduction in the average number of myofibroblasts in the siRNA-p65 group compared with that in the siRNA-neg group. Thus, intra-articular p65-siRNA injection could attenuate fibroblast activation and fibrosis-related protein production, suppress periarticular tissue fibrosis, and prevent joint contracture by downregulating the NF-κB p65 pathway. Statement of clinical significance: Intra-articular injection of p65-siRNA could reduce myofibroblast proliferation and fibrosis-related protein expression by downregulating the NF-κB p65 pathway, inhibit periarticular tissue fibrosis, and prevent joint adhesion, which represents a potential therapy in the prevention of joint fibrosis following traumatic injury.

Keywords: NF‐κB; RelA/p65; fibrosis; joint contracture; siRNA.

Abstract

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