Effect of Gubitongxiao granules in a mouse model of steroid-induced necrosis of the femoral head / 中国组织工程研究

BACKGROUND:Glucocorticoids can inhibit the expression of hub genes in the parathyroid hormone type Ⅰ receptor(PTH1R)/protein kinase A(PKA)signaling axis and interfere with the osteogenic and angiogenic differentiation of bone marrow mesenchymal stem cells,leading to the disruption of blood supply in bone and bone tissue structures.Previous studies of the research team showed that Gubitongxiao granules can induce blood vessel formation and inhibit osteoblast apoptosis,which has a certain effect on the prevention and treatment of steroid-induced femoral head necrosis. OBJECTIVE:To observe the therapeutic effect of Gubitongxiao granules in a mouse model of steroid-induced femoral head necrosis,and to explore its mechanism from the PTH1R/PKA signaling axis. METHODS:An animal model of steroid-induced necrosis of the femoral head was established by intraperitoneal injection of lipopolysaccharide and gluteal muscle injection of prednisolone acetate.After identification by nuclear magnetic resonance method,60 mice that were successfully modeled were divided into model group,Gubitongxiao granule group and Tongluo Shenggu capsule group,with 20 mice in each group.Another 12 normal mice were used as control group.The corresponding groups were intragastrically given the corresponding drugs for 12 weeks,and then the samples were taken under anesthesia.Histomorphology of femoral head samples was observed by hematoxylin-eosin staining.Enzyme-linked immunosorbent assay was used to detect the serum levels of bone alkaline phosphatase,type Ⅰ amino-terminal extension peptide,parathyroid hormone,osteocalcin and alkaline phosphatase.Western blot and RT-qPCR were used to detect PTH1R,PKA,myocyte enhancer factor 2,sclerostin and guanylate-binding protein activity-stimulating peptide at protein and gene expression levels,respectively. RESULTS AND CONCLUSION:Gubitongxiao granules may reduce the serum PTH level in mice,inhibit the activation of the PTH1R/PKA signal axis,further up-regulate the protein expressions of sclerostin and myocyte enhancer factor 2,and increase the levels of bone alkaline phosphatase,type Ⅰ amino-terminal extension peptide,osteocalcin and alkaline phosphatase in mice,thus improving femoral head necrosis,which is comparable to the intervention effect of Tongluo Shenggu capsules.It is speculated that Gubitongxiao granules may prevent and treating hormonal femoral head necrosis by regulating the PTH1R/PKA signaling axis.

Effect of losartan on expression of PTHrP and PTH1R induced by high glucose in NRK-52E cells / 中华内分泌代谢杂志

To study the effect of losartan on high glucose up-regulated expression of parathyroid hormone-related peptide (PTHrP) and its type 1 receptor (PTH1R) in NRK-52E cells. The expression of PTHrP and PTH1R were detected by RT-PCR and Western blot in control group (0 mmol/L glucose) ,normal glucose group (5 mmol/ L glucose) ,moderately high glucose group (16.7 mmol/L glucose) ,high glucose group (25 mmol/L glucose) ,and after intervention by 10 μmol/L losartan for 72 h (only Western blot). The expression of PTHrP and PTH1R were up-regulated by high glucose (PTHrP mRNA : 0. 66 ± 0.08, 0. 84 ± 0. 13,1.57 ± 0. 15, and 1.73 ± 0.21 ; PTHrP protein :0.63±0.12,0.68±0.06,1.02±0. 11, and 1.04±0.08 ; PTH1R mRNA :0.26±0. 08,0.28±0.07,2.35± 0. 10,and 2.47±0. 05 ; PTH1R protein:0. 88±0. 05,0. 87±0. 10, 1.05±0. 11, and 1.12±0. 09) ,and losartan inhibited the effects of high glucose (PTHrP 0.74±0. 15, PTH1R 0.98±0.06, both P<0.01). The results suggest that losartan could inhibit the expression of FTHrP and PTH1R induced by high glucose in NRK-52E.