Duhuo Jisheng Decoction Disassembled Prescription inhibits chondrocyte inflammatory response  throughbased on Wnt/beta-catenin signaling pathway

doi:10.3969/j.issn.2095-4344.2885

Abstract

Abstract:

BACKGROUND: ThePrevious studies have shown that the Wnt/β-catenin signaling pathway plays an important role in the pathogenesis and prognosis of chondrocyte inflammation, and the effective inhibition of chondrocyte inflammatory response byDuhuo Jisheng Decoction Disassembled Prescription remains to be further studied.

OBJECTIVE: To investigate whether Duhuo Jisheng Decoction Disassembled Prescription can inhibit the inflammatory response of chondrocytes induced by lipopolysaccharide via the Wnt/β-catenin signaling pathway.
METHODS: Male Sprague-Dawley rats were used to obtain knee joint chondrocytes by mechanical-type II collagenase digestion. The morphology and structure of chondrocytes were observed by inverted phase contrast microscope. Chondrocytes were identified by type II collagen immunohistochemistry. Duhuo Jisheng Decoction Disassembled Prescription with different mass concentrations (300, 400, 500 mg/L) was used to deal with the chondrocyte inflammation model induced by lipopolysaccharide (10 μg/L). After 8 hours of intervention, the ELISA method was used to determine the expression of interleukin-1β and tumor necrosis factor-α in the cell culture fluid to determine the intervention concentration of Duhuo Jisheng Decoction Disassembled Prescription. The second-generation chondrocytes were divided into four groups, followed by culture with normal medium in blank group, 10 μg/L lipopolysaccharide in model group, 10 μg/L lipopolysaccharide plus 400 mg/L Duhuo Jisheng Decoction Disassembled Prescription in 400 mg/L Duhuo Jisheng Decoction Disassembled Prescription group, and 10 μg/L lipopolysaccharide plus 10 mg/L Dickkopf-1 in inhibitor group. After 8 hours of intervention, the expression levels of β-catenin, Wnt-4, Frizzled-2, GSK-3β and CKI-ε were detected by western blot.
RESULTS AND CONCLUSION: Chondrocyte identification results showed that the cytoplasm of chondrocytes was brown-yellow, with the typical characteristics of chondrocytes. ELISA results showed that the expression levels of interleukin-1β and tumor necrosis factor-α were higher in the model group than in the blank group, and the expression levels of interleukin-1β and tumor necrosis factor-α were significantly reduced in the 400 mg/L Duhuo Jisheng Decoction Disassembled Prescription group compared with the model group. Therefore, the intervention concentration of Duhuo Jisheng Decoction Disassembled Prescription was determined to be 400 mg/L. The results of western blot showed that the expression levels of Wnt-4, Frizzled-2, β-catenin and CKI-ε in the model group were higher than those in the blank group, whereas the expression of GSK-3β protein was lower than that in the blank group. The expression levels of Wnt-4, Frizzled-2, β-catenin and CKI-ε proteins in the Duhuo Jisheng Decoction Disassembled Prescription group and the inhibitor group were lower than those in the model group, whereas the protein expression level of GSK-3β was higher than that in the model group. All the findings indicate that Duhuo Jisheng Decoction Disassembled Prescription can inhibit lipopolysaccharide-induced chondrocyte inflammation by regulating the Wnt/β-catenin signaling pathway.

Key words: Duhuo Jisheng Decoction Disassembled Prescription, chondrocytes, Wnt/β-catenin signaling pathway, Wnt-4, Frizzled-2, β-catenin, GSK-3β, CKI-ε

CLC Number:

Li Hui, Ma Yuhuan, Xu Limei, Wang Shengjie, Xu Yunteng, He Xiaojuan, Jia Liangliang, Zeng Jianwei, Wang Lili, Li Xihai, Ye Hongzhi. Duhuo Jisheng Decoction Disassembled Prescription inhibits chondrocyte inflammatory response throughbased on Wnt/beta-catenin signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(35): 5589-5594.

Figures/Tables 4

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