Effect of Tongluo Shenggu Capsule on toll-like receptor 4 signaling pathway in rats with steroid-induced osteonecrosis of the femoral head

doi:10.12307/2021.113

Abstract

Abstract:
BACKGROUND: Early pharmacological studies have found that Tongluo Shenggu Capsule has various mechanisms for repairing steroid-induced osteonecrosis of the femoral head (SANFH), but its influence on the toll-like receptor 4 (TLR4) signaling pathway has not been documented.
OBJECTIVE: To investigate the molecular mechanism of Tongluo Shenggu Capsule in SANFH treatment based on the TLR4 signaling pathway.
METHODS: A total of 60 female Sprague-Dawley rats were randomly divided into a normal group (20 rats) and a model group (40 rats). Endotoxin combined with methylprednisolone was used to establish the SANFH rat model. The normal group was injected with normal saline at the same volume. After modeling, 5 rats from the normal group and 10 rats from the model group were selected for observation under light microscope. The 30 rats successfully modeled were randomly divided into two groups: a hormone group and an intervention group with 15 rats in each group. The intervention group was given Tongluo Shenggu Capsule solution 0.4 g/kg once a day for 4 weeks. The hormone group and the normal group were given the same dose of normal saline for 4 weeks, once a day. The hormone and normal groups were given the same amount of normal saline once a day for 4 weeks. After the end of intervention, bilateral femoral heads of rats in each group were taken for morphological observation and pathological detection. Expressions of TLR4, MyD88, tumor necrosis factor-β and nuclear factor-κB proteins were detected by western blot.
RESULTS AND CONCLUSION: The femoral head of rats in the normal group had a lustrous and smooth appearance without deformation or collapse. The femoral head of rats in the hormone group had a dull, purple appearance and a large area, while the femoral head of rats in the intervention group tended to be normal. Hematoxylin-eosin staining results showed that in the hormone group, trabeculae of the femoral head were sparse and even broken, bone nuclei were solidified, dissolved and disappeared, and the number of empty bone lacunae was the largest. Compared with the normal group and the intervention group, the difference in the rate of empty bone lacunae was significant (P < 0.05). In the intervention group, the bone trabeculae were large and regular, and the cell morphology was mostly normal, which was improved compared with the hormone group. Protein levels of nuclear factor-κB, tumor necrosis factor-β, MyD88 and TLR4 detected by western blot were significantly lower in the normal group and the intervention group than in the hormone group (P < 0.05). There was no significant difference in TLR4 and MyD88 expression between the intervention group and the normal group (P > 0.05). In conclusion, the mechanism of Tongluo Shenggu Capsule in SANFH treatment may be related to inhibition of MyD88-dependent TLR4 signaling pathway.

Key words: bone, osteonecrosis of the femoral head, steroid-induced osteonecrosis of the femoral head, Tongluo Shengu Capsule, TLR4, signaling pathway, rat

CLC Number:

Liu Jinfu, Zeng Ping, Nong Jiao, Fan Siqi, Qian Xiaofen, Feng Chengqin, Lei Zhiqiang, Luo Jun, Huang Jiaxing. Effect of Tongluo Shenggu Capsule on toll-like receptor 4 signaling pathway in rats with steroid-induced osteonecrosis of the femoral head[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(26): 4150-4155.

Figures/Tables 3

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