The mechanism by which Zhuanggu Jianxi Decoction inhibits inflammatory response of human synovial macrophages

doi:10.12307/2026.111

Abstract

Abstract: BACKGROUND: Synovial macrophages are one of the main cell types in the synovial tissue and are the primary cell type causing synovial inflammation. As a current research hotspot in knee osteoarthritis inflammation-related studies, our research group has previously conducted a series of targeted studies, preliminarily verifying through rabbit synovial tissue and cell levels that the prescription of Zhuanggu Jianxi Decoction can alleviate synovial inflammation by regulating the liver X receptors (LXRs)/nuclear factor- kappaB (NF-κB) pathway. This study takes human synovial macrophages as the research object, which is closer to the real state of patients with knee osteoarthritis, with the aim of further clarifying the mechanism by which Zhuanggu Jianxi Decoction reduces synovial inflammation.
OBJECTIVE: To explore the effects and potential molecular mechanism of Zhuanggu Jianxi Decoction-containing serum on lipopolysaccharide-induced synovial inflammation in human synovial macrophages.
METHODS: (1) Twelve 3-month-old New Zealand white rabbits were used to prepare the drug-containing serum of Zhuanggu Jianxi Decoction and blank serum. (2) Human synovial macrophages were induced to establish an inflammatory injury model by lipopolysaccharide (1.0 μg/mL). The cells were randomly divided into the model group (10% blank serum), the drug-containing serum group (10% drug-containing serum), the LXRα blocker group (10% drug-containing serum + 5 μg LXRα blocker), the nuclear receptor co-repressor (N-CoR) blocker group (10% drug-containing serum + 5 μg N-CoR blocker), and the normal group (10% blank serum). All the groups were intervened for 24 hours. (3) Western blot and RT-qPCR were used to detect the expression of LXRα, N-CoR, P50, and P65 proteins and mRNAs in each group of cells, and ELISA was used to detect the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant.
RESULTS AND CONCLUSION: (1) Compared with the normal group, the model group showed significantly decreased LXRα and N-CoR protein/mRNA expressions (P < 0.01), while P50/P65 protein/mRNA expressions and the levels of interleukin-1β/tumor necrosis factor-α were significantly elevated (P < 0.01). (2) Compared with the model group, drug-containing serum significantly upregulated LXRα/N-CoR expressions at protein and mRNA levels and downregulated P50/P65 expressions at protein and mRNA levels as well as the levels of interleukin-1β/tumor necrosis factor-α (P < 0.01). Compared with the model group, both blocker groups showed a reduction in P50/P65 expressions at protein and mRNA levels and the levels of interleukin-1β/tumor necrosis factor-α (P < 0.01, P < 0.05). (3) Compared with the drug-containing serum group, both blocker groups exhibited significantly lower the protein and mRNA expressions of LXRα/N-CoR and higher protein and mRNA expressions of P50/P65 and higher levels of interleukin-1β/tumor necrosis factor-α (P < 0.01, P < 0.05). To conclude, Zhuanggu Jianxi Decoction-containing serum alleviates synovial macrophage inflammation and then prevent/treat knee osteoarthritis by upregulating the expression of LXRα/N-CoR protein and mRNA, downregulating the expression of P50/P65 proteins and mRNA, inhibiting the transmission of the LXRs/NF-κB signaling pathway, and reducing the secretion of downstream inflammatory markers (interleukin-1β and tumor necrosis factor-α).

Key words: knee osteoarthritis, synovial inflammation, Zhuanggu Jianxi Decoction, human synovial macrophages, LXRs, NF-κB, signaling pathway

CLC Number:

Peng Siwei, Yang Ruifang, Chen Xiaohua, Zheng Zhuoming, Chen Peng, Xiao Yan, Su Youxin, Guo Jiemei. The mechanism by which Zhuanggu Jianxi Decoction inhibits inflammatory response of human synovial macrophages[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(17): 4400-4406.

Figures/Tables 1

2.1 各组人滑膜巨噬细胞中 LXRα、N-CoR、P50及P65蛋白表达如图1，2所示，与正常组相比，模型组人滑膜巨噬细胞中LXRα和N-CoR蛋白表达显著降低(P < 0.01)，P50和P65蛋白表达显著升高(P < 0.01)。与模型组相比，含药血清组LXRα与N-CoR蛋白表达均显著升高(P < 0.01)，P50和P65蛋白表达均显著降低(P < 0.01)；LXRα阻滞剂组及N-CoR阻滞剂组P50和P65蛋白表达降低(P < 0.01，P < 0.05)。与含药血清组相比，LXRα阻滞剂组及N-CoR阻滞剂组LXRα及N-CoR蛋白表达显著降低(P < 0.01)，P50和P65蛋白表达升高(P < 0.01，P < 0.05)。 2.2 各组人滑膜巨噬细胞中LXRα、N-CoR、P50及P65 mRNA表达如图3所示，与正常组相比，模型组人滑膜巨噬细胞中LXRα和N-CoR mRNA表达均显著降低(P < 0.01)，P50和P65 mRNA表达均显著升高(P < 0.01)。与模型组相比，含药血清组LXRα和N-CoR mRNA表达均显著升高(P < 0.01)，P50和P65 mRNA表达均显著降低(P < 0.01)；LXRα阻滞剂组P50和P65 mRNA表达降低(P < 0.05)、N-CoR阻滞剂组P50和P65 mRNA表达显著降低(P < 0.01)；N-CoR阻滞剂组LXRα mRNA表达显著升高(P < 0.01)。与含药血清组相比，LXRα阻滞剂组及N-CoR阻滞剂组LXRα和N-CoR mRNA表达均显著降低(P < 0.01)，P50和P65 mRNA表达均显著升高(P < 0.01)。 2.3 各组人滑膜巨噬细胞上清液白细胞介素1β、肿瘤坏死因子α水平如图4所示，与正常组相比，模型组中白细胞介素1β、肿瘤坏死因子α水平均显著升高(P < 0.01)。与模型组相比，含药血清组、LXRα阻滞剂组及N-CoR阻滞剂组白细胞介素1β水平均显著降低(P < 0.01)，肿瘤坏死因子α水平显著降低(P < 0.01，P < 0.05)。与含药血清组比较，LXRα阻滞剂组和N-CoR阻滞剂组白细胞介素1β、肿瘤坏死因子α水平显著升高(P < 0.01，P < 0.05)。"

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