Li Jun, Peng Youjian
Department of Stomatology, Renmin Hospital of Wuhan University, Wuhan, China.
J Int Med Res. 2020 Apr;48(4):300060519851641. doi: 10.1177/0300060519851641. Epub 2019 Dec 29.
To investigate the effects of the flavonoid, puerarin, on osteogenic differentiation of human periodontal ligament stem cells (PDLSCs).
Human PDLSCs were isolated from patients undergoing orthodontic treatment, and the cell surface markers CD146, CD34, CD45, and STRO-1 were identified by immunofluorescence. Cell proliferation was detected by MTT assay; alkaline phosphatase (ALP) activity was measured, and calcium deposition was detected by alizarin red staining. PCR was then used to detect the distributions of , , , and , genes related to osteogenic differentiation.
Staining was positive for cytokines CD146, CD34, CD45, and STRO-1 in the experimental group; staining was also positive for silk protein, but negative for keratin. After 7 days of culture, exposure to puerarin significantly promoted the level of intracellular ALP; increased puerarin concentration led to increased intracellular ALP. Red mineralized nodules appeared upon exposure to puerarin and the number of nodules was concentration-dependent. PCR analysis revealed that , , , and expression levels increased as puerarin concentration increased.
Exposure to puerarin can promote proliferation and ALP activity in human PDLSCs, thus promoting both molecular and osteogenic differentiation; these findings may provide a theoretical basis for the clinical treatment of periodontal disease with puerarin.
研究黄酮类化合物葛根素对人牙周膜干细胞（PDLSCs）成骨分化的影响。
从接受正畸治疗的患者中分离出人牙周膜干细胞，通过免疫荧光鉴定细胞表面标志物CD146、CD34、CD45和STRO-1。采用MTT法检测细胞增殖；测定碱性磷酸酶（ALP）活性，并用茜素红染色检测钙沉积。然后用PCR检测与成骨分化相关的、、、和基因的分布。
实验组细胞因子CD146、CD34、CD45和STRO-1染色呈阳性；丝蛋白染色也呈阳性，但角蛋白染色呈阴性。培养7天后，葛根素处理显著提高了细胞内ALP水平；葛根素浓度增加导致细胞内ALP增加。葛根素处理后出现红色矿化结节，且结节数量呈浓度依赖性。PCR分析显示，随着葛根素浓度增加，、、、和的表达水平升高。
葛根素处理可促进人牙周膜干细胞的增殖和ALP活性，从而促进分子和成骨分化；这些发现可能为葛根素临床治疗牙周病提供理论依据。
