Effect of KH-BaRoKer-SeongJangTang based on traditional medicine theory on longitudinal bone growth

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  • ABSTRACT

    KH-BaRoKer-SeongJangTang (KBS) is a recently developed formulation by using traditional drugs considering traditional medical theory of Oriental books such as ShinNongBonChoGyeong and JuRye, which has been used to improve the growth of child in Korea. Although KBS is usually prescribed to many children who are in retard for their age, its pharmacological effects have not been fully understood in experimental models. The aim of this study was to evaluate the effects of KBS on bone growth. Growth plate thickness and bone parameters such as bone volume/tissue volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), connection density (Conn.D), and total porosity were analyzed by means of microcomputed tomography. Serum insulin-like growth factor-I (IGF-I) levels were measured by enzyme-linked immunosorbent assay. Hepatic IGF-I mRNA expression was analyzed by real-time polymerase chain reaction. Phosphorylation of signal transducer and activator of transcription5 (STAT5) was investigated using Western blot analysis and immunohistochemistry. The thickness of growth plate was increased by KBS. BV/TV, Tb.Th, TbN, Conn.D, and total porosity were improved by KBS. Hepatic IGF-I mRNA and serum IGF-I levels were elevated by KBS. Phosphorylation of STAT5 was increased with administration of KBS. These results suggest that KBS would be helpful to children who are in retard for their age through the elevation of IGF-I.


  • KEYWORD

    growth plate , bone parameters , insulin-like growth factor-I , signal transducer and activator of transcription5

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  • [Table 1.] Composition of Experimental Diets.a
    Composition of Experimental Diets.a
  • [Table 2.] The Ratio of the Components in KBS
    The Ratio of the Components in KBS
  • [Fig. 1.] Effect of KBS on tibial growth plate thickness. (A) Representative 3D μCT images of knee joint showing growth plate. (B) The thickness of excised bone growth plate was determined on five points using the Sky Scan 1076. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Arg-administered group; Glu, low protein diet + Glu-administered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p< 0.05; significantly different from the PEM value.
    Effect of KBS on tibial growth plate thickness. (A) Representative 3D μCT images of knee joint showing growth plate. (B) The thickness of excised bone growth plate was determined on five points using the Sky Scan 1076. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Arg-administered group; Glu, low protein diet + Glu-administered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p< 0.05; significantly different from the PEM value.
  • [Fig. 2.] Effect of KBS on trabecular bone properties. BMD and trabecular parameters of excised bone at the tibia were determined. (A) BMD using 2D μCT. (B-F) Quantified parameters of proximal tibia using 3D μCT. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Argadministered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p < 0.05; significantly different from the PEM value.
    Effect of KBS on trabecular bone properties. BMD and trabecular parameters of excised bone at the tibia were determined. (A) BMD using 2D μCT. (B-F) Quantified parameters of proximal tibia using 3D μCT. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Argadministered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p < 0.05; significantly different from the PEM value.
  • [Fig. 3.] Effect of KBS on the regulation of IGF-I. (A) IGF-I levels in the serum were measured with the ELISA method. (B) The expression levels of IGF-I mRNA were evaluated by the real-time PCR method. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBSadministered group; Arg, low protein diet + Arg-administered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p < 0.05; significantly different from the PEM value.
    Effect of KBS on the regulation of IGF-I. (A) IGF-I levels in the serum were measured with the ELISA method. (B) The expression levels of IGF-I mRNA were evaluated by the real-time PCR method. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBSadministered group; Arg, low protein diet + Arg-administered group. Each datum represents the mean ± SEM of three independent experiments. #p < 0.05; significantly different from the CON value. *p < 0.05; significantly different from the PEM value.
  • [Fig. 4.] Effect of KBS on the phosphorylation of STAT5. (A) Liver tissues of each mouse were homogenized and analysed for pSTAT5 and STAT5 by Western blot analysis using specific anti-pSTAT5 and STAT5 antibodies as described in the experimental procedures. (B) pSTAT5 was analyzed by immunohistochemistry in the tibia. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Arg-administered group.
    Effect of KBS on the phosphorylation of STAT5. (A) Liver tissues of each mouse were homogenized and analysed for pSTAT5 and STAT5 by Western blot analysis using specific anti-pSTAT5 and STAT5 antibodies as described in the experimental procedures. (B) pSTAT5 was analyzed by immunohistochemistry in the tibia. CON, adequate protein diet + DW-administered group; PEM, low protein diet + DW-administered group; KBS, low protein diet + KBS-administered group; Arg, low protein diet + Arg-administered group.