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. 2022 May 26:13:881057.
doi: 10.3389/fphar.2022.881057. eCollection 2022.

Total Flavonoids of Rhizoma Drynariae Ameliorate Bone Growth in Experimentally Induced Tibial Dyschondroplasia in Chickens via Regulation of OPG/RANKL Axis

Tingting Xu  1 , Jingjing Zheng  1 , WeiXing Jin  2 , Lu Li  1 , Luxi Lin  1 , Aftab Shaukat  3 , Chaodong Zhang  1 , Qinqin Cao  1 , Muhammad Ashraf  4 , Shucheng Huang  1
Affiliations

Total Flavonoids of Rhizoma Drynariae Ameliorate Bone Growth in Experimentally Induced Tibial Dyschondroplasia in Chickens via Regulation of OPG/RANKL Axis

Tingting Xu et al. Front Pharmacol. .

Erratum in

Abstract

Background: Rhizoma Drynariae, traditional Chinese herb, is widely used to treat and prevent bone disorders. However, experimental evidence on the use of Rhizoma Drynariae extract, total flavonoids of Rhizoma Drynariae (TFRD) to treat tibial dyschondroplasia (TD) in chickens and its underlying mechanisms have not been investigated. Purpose: To evaluate the therapeutic effect of TFRD on leg disease caused by TD and elucidate its mechanisms in modulating the bone status. Methods: Thiram-induced chicken TD model has been established. The tibia status was evaluated by analyzing tibia-related parameters including tibial weight, tibial length and its growth plate width and by performing histopathological examination. The expression of tibial bone development-related genes and proteins was confirmed by western blotting and qRT-PCR. Results: The results showed that administration of TFRD mitigated lameness, increased body weight, recuperated growth plate width in broilers affected by TD and the increase of tibia weight and tibia length is significantly positively correlated with body weight. Compared with the TD group broilers, 500 mg/kg TFRD evidently reduced the damage width of the growth plate and improved its blood vessel distribution by elevating the gene expression levels of BMP-2 and Runx2 and OPG/RANKL ratio. Furthermore, correlation analysis found that the damage width of the growth plate was negatively correlated with the expression levels of BMP-2 and OPG. Conclusion: The present study revealed that TFRD could promote the bone growth via upregulating OPG/RANKL ratio, suggesting that TFRD might be a potential novel drug in the treatment of TD in chickens.

Keywords: Chinese herbal medicine; bone development; leg disease; tibial dyschondroplasia; total flavonoids of Rhizoma Drynariae.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AL declared a shared affiliation with the author AS to the handling editor at the time of review.

Figures

FIGURE 1
FIGURE 1
Effects of TFRD on tibial growth in TD broilers. (A) The tibial of chickens. (B) Body weight of chickens. (C) Tibia weight. (D) Tibia weight index. (E) Tibia length. (F) Tibia growth plate width. (G) Tibia growth plate index. (H) The pearson correlation analysis between tibia weight and body weight. (I) The pearson correlation analysis between tibia length and body weight. (J) The pearson correlation analysis between tibia growth plate width and body weight. The results are represented as mean ± SD. a, b, c, and d represent significant differences between groups (p < 0.05).
FIGURE 2
FIGURE 2
Effects of TFRD on growth plate of tibia bone in TD broilers. (A) The morphology result of tibial growth plate. GP, growth plate; TDL, tibial dyschondroplasias lesion. (B) The HE staining observation result of tibial growth plate. Scar bar = 1,000 µm.
FIGURE 3
FIGURE 3
Effects of TFRD on vascular invasion of tibial growth plate in TD broilers. HE staining was used to observe the vascular infiltration of growth plates in each group. The black arrow points to the blood vessel. Scar bar = 100 µm.
FIGURE 4
FIGURE 4
Effects of TFRD on gene expression levels of BMP-2/Runx2 and OPG/RANKL in tibial growth plate in TD broilers. (A–D) Principal component analysis (PCA) of the BMP-2, Runx2, OPG and RANKL gene expressions with an unsupervised pattern recognition method. (E) The mRNA expression of genes BMP-2, Runx2, OPG, RANKL and the ratio of OPG/RANKL at day 7, 14 and 21. (F) The heat map shows the mRNA levels of the bone-related genes. The results are represented as mean ± SD. a, b, c, and d represent significant differences between groups (p < 0.05).
FIGURE 5
FIGURE 5
Effects of LTFRD on OPG/RANKL protein level of tibial growth plate in TD broilers. (A) The gray scale analysis of OPG, RANKL and β-ACTIN. (B) The protein level of OPG. (C) The protein level of RANKL. (D) The protein level of OPG/RANKL The results are represented as mean ± SD. a, b, c, and d represent significant differences between groups (p < 0.05).
FIGURE 6
FIGURE 6
Correlation heat map analysis of tibial growth parameters and osteogenesis-related biological indicators in broilers. Note: The different rectangles are colored based on the Pearson correlation coefficients between tibia growth performance expression levels and bone formation-related indices. The intensity of color represents the degree of correlation, red represents positive correlation, blue represents negative correlation. *indicates significant correlation (p < 0.05), ***indicates highly significant correlation (p < 0.001).
FIGURE 7
FIGURE 7
The schematic diagram of the protective mechanism of TFRD on TD broilers by regulating the OPG/RANKL expression.
See this image and copyright information in PMC

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