Intervention Effects of Salvia Miltiorrhiza Extract on Human Periodontal Ligament Fibroblasts and Its Effect on CTGF and ALP Levels

OBJECTIVE To study the effects of Salvia miltiorrhiza extract on the levels of connective tissue growth factor (CTGF) and alkaline phosphatase(ALP) in human periodontal ligament fibroblasts. METHODS Two first premolars extracted by orthodontic treatment in Hebei Hospital of Traditional Chinese Medicine were put into aseptic α-MEM medium containing penicillin and streptomycin immediately after tooth extraction. Human periodontal ligament fibroblasts were isolated and cultured. The fifth generation of human periodontal ligament fibroblasts were used for follow-up test, which were divided into blank group, low and high concentration group of Salvia miltiorrhiza extract. The blank group was added with 100 μL basic medium, low concentration group of Salvia miltiorrhiza extract was added to 1 μmol·L^-1 Salvia miltiorrhiza extract solution, and 2 μmol·L^-1 Salvia miltiorrhiza extract solution was added to high concentration group of Salvia miltiorrhiza extract. Cell proliferation, apoptosis, osteogenic differentiation and expression of phosphorylated c-Jun amino terminal kinase(p-JNK), CTGF, bone morphogenetic protein-2(BMP-2), osteogenic specific transcription factor(Runx-2) and ALP were measured. RESULTS The proliferation rate and ALP activity of high concentration group of Salvia miltiorrhiza extract in 24, 48, 72 h were higher than that of low concentration group of Salvia miltiorrhiza extract(P<0.05). The apoptosis rate of high concentration group of Salvia miltiorrhiza extract in 24, 48, 72 h was lower than that of low concentration group of Salvia miltiorrhiza extract(P<0.05). The expression levels of p-JNK, CTGF, BMP-2, Runx-2 and ALP in the high concentration group of Salvia miltiorrhiza extract were all higher than those in the low concentration group of Salvia miltiorrhiza extract(P<0.05). CONCLUSION Salvia miltiorrhiza extract by acting on JNK pathway, raise the CTGF expression, promote the periodontal ligament fibroblasts proliferation, through the regulation of BMP-2, Runx-2 expression, increase ALP activity, promote the periodontal ligament fibroblasts osteogenesis differentiation, and has a certain concentration dependence.