Anti-Osteoporosis Effect of Heme on Osteoblast Pre-Cells and Rat Model

Abstract

Objectives: Osteoporosis is a metabolic bone disease caused by various factors. As a prosthetic group of haemoglobin, heme can promote the formation of blood vessels and the regeneration of related cells by increasing the proliferation level of endothelial cells and reticulocytes. This study observed the anti-osteoporosis effect of heme on preosteoblast (POB) cells and rat models. Methods: Heme mimics were transfected into POB cells of patients in stable culture, and the effect of heme transfection on the activity of POB cells was detected via the cell scratch test and cell migration assay. 45 SD female rats were randomly divided into group A, the rats were only treated with surgery (n = 15); group B, the rats were given 30 mg/kg of heme chloride by tail vein injection on the 1st, 7th, and 14th days after surgery (n = 15); group C, rats were injected with the same amount of saline in the tail vein (n = 15). The mRNA and protein expressions of alkaline phosphatase (ALP), bone-specific alkaline phosphatase (B-ALP), and type I collagen C-terminal peptide (CTX) in rats serum were detected via western blot and real-time PCR (real-time polymerase chain reaction). The expressions of IL-6 and TNF-α in rats serum were detected via Elisa. The pathological changes in the morphology of distal femur were observed by Hematoxylin-eosin staining. Results: After transfected with heme mimics, the proliferation, migration and invasion ability of the patient’s cells increased significantly, and the apoptosis rate of the cells decreased significantly. The relative expression levels of ALP and CTX mRNA and protein in serum of osteoporosis patients and rat significantly decreased 24 hours after transfection of heme mimics (p < 0.05), while B-ALP significantly increased. The expression of IL-6 and TNF-α in the rat model of osteoporosis group significantly increased (p < 0.05), but after transfection with heme mimic, the expression of IL-6 and TNF-α in the rat reduced significantly. HE showed that, after transfection, the femoral trabeculae were substantially broader and thicker, and the number also increased significantly. The number of trabecular fractures and fat cells was decreased, the trabecular bones were smoother and close to the control group. Conclusion: Heme can significantly promote the proliferation, migration and invasion of POB cells in the body, and also can achieve the anti-osteoporosis effect by adjusting the expression of ALP, B-ALP, and CTX, IL-6 and TNF-α in osteoporosis model of rat.