Effect of Wulao Qisun Prescription on Proliferation and Osteogenic Differentiation of AS Fibroblasts by Regulating Wnt/β-catenin Signaling Pathway
10.13422/j.cnki.syfjx.20241406
- VernacularTitle:五劳七损方调控Wnt/β-catenin信号通路对AS成纤维细胞增殖及成骨分化的影响
- Author:
Juanjuan YANG
1
;
Ping CHEN
2
;
Haidong WANG
2
;
Zhendong WANG
1
;
Haolin LI
1
;
Zhimin ZHANG
1
;
Yuping YANG
1
;
Weigang CHENG
1
;
Jin SU
1
;
Jingjing SONG
1
;
Dongsheng LU
1
Author Information
1. Clinical College of Chinese Medicine，Gansu University of Chinese Medicine，Lanzhou 730000，China
2. Gansu Provincial Hospital of Traditional Chinese Medicine，Lanzhou 730050，China
- Publication Type:Journal Article
- Keywords:
Wulao Qisun prescription;
ankylosing spondylitis;
Wnt/β-catenin signaling pathway;
fibroblasts;
osteogenic differentiation
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(2):67-73
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis （AS）. MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group， low drug-containing serum group， medium drug-containing serum group， high drug-containing serum group， and positive drug group. After drug intervention， cell proliferation was measured using the cell counting kit-8 （CCK-8） assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase （ALP） activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin （OCN）， osteopontin （OPN）， and runt-related transcription factor 2 （Runx2） was detected by Western blot. The mRNA expression levels of Wnt5a， β-catenin， and Dickkopf-1 （DKK-1） were measured by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsCompared with the blank group， each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression （P<0.05）. The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression （P<0.05， P<0.01）， with the positive drug group showing the most pronounced effect （P<0.01）. The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins （P<0.05， P<0.01）， while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN， OPN， and Runx2 proteins （P<0.05， P<0.01）. ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts， thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression， further inhibiting the transcription of downstream target genes.
