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没食子酸表没食子儿茶素酯通过调控 PI3K/Akt 信号通路影响人牙周膜细胞和牙槽骨细胞的增殖及分化。

Epigallocatechin gallate affects the proliferation of human alveolar osteoblasts and periodontal ligament cells, as well as promoting cell differentiation by regulating PI3K/Akt signaling pathway.

机构信息

The Affiliated Stomatologic Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

Department of Stomatology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.

出版信息

Odontology. 2021 Jul;109(3):729-740. doi: 10.1007/s10266-021-00597-1. Epub 2021 Mar 6.

Abstract

Human periodontal ligament cells (hPDLCs) and human alveolar osteoblasts (hAOBs) play pivotal roles in periodontium. The regulatory effects of epigallocatechin gallate (EGCG) on hPDLCs and hAOBs remained unclear. This study probed into the functions of EGCG treating periodontal diseases. Cultured hAOBs and hPDLCs were passaged and observed by microscopic examination, and alkaline phosphatase (ALP) and immumohistochemical staining were performed for verification. hAOBs and hPDLCs were treated with EGCG and LY294002 + EGCG, then the proliferation of the two cells was assayed by MTT. Mineralization of the treated hAOBs and hPDLCs was detected by ALP activity experiment and Alizarin Red S staining kit. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were performed for the detection of the expressions of differentiation-related mRNAs and PI3K/Akt signaling pathway-related proteins in the two cells. The third passage of hAOBs mainly showed triangle shape and were positive by ALP staining. hPDLCs in passage 3 adhered to the wall in spiral or radial pattern with positively stained vimentin and negatively stained keratin. Cell proliferation and ALP activity of the hAOBs and hPDLCs were increased by EGCG treatment. The mineralized nodules and expressions of differentiation-related mRNAs, the phosphorylation of PI3K and Akt of the hAOBs and hPDLCs were promoted by EGCG treatment, while the effects of LY294002 treatment were opposite to EGCG treatment. Epigallocatechin gallate affected the proliferation and differentiation of hAOBs and hPDLCs through regulating PI3K/Akt signaling pathway.

摘要

人牙周韧带细胞(hPDLCs)和人牙槽骨细胞(hAOBs)在牙周组织中发挥着关键作用。表没食子儿茶素没食子酸酯(EGCG)对 hPDLCs 和 hAOBs 的调节作用尚不清楚。本研究探讨了 EGCG 治疗牙周病的作用。通过显微镜观察传代培养的 hAOBs 和 hPDLCs,并进行碱性磷酸酶(ALP)和免疫组化染色验证。用 EGCG 和 LY294002+EGCG 处理 hAOBs 和 hPDLCs,然后通过 MTT 测定两种细胞的增殖。通过 ALP 活性实验和茜素红 S 染色试剂盒检测处理后的 hAOBs 和 hPDLCs 的矿化。通过定量实时聚合酶链反应(qRT-PCR)和 Western blot 检测两种细胞中分化相关 mRNAs 和 PI3K/Akt 信号通路相关蛋白的表达。第 3 代 hAOBs 主要呈三角形,ALP 染色阳性。第 3 代 hPDLCs 呈螺旋状或放射状贴壁生长,波形蛋白染色阳性,角蛋白染色阴性。EGCG 处理可增加 hAOBs 和 hPDLCs 的细胞增殖和 ALP 活性。EGCG 处理可促进 hAOBs 和 hPDLCs 的矿化结节形成和分化相关 mRNAs 的表达,以及 PI3K 和 Akt 的磷酸化,而 LY294002 处理的效果则与 EGCG 处理相反。表没食子儿茶素没食子酸酯通过调节 PI3K/Akt 信号通路影响 hAOBs 和 hPDLCs 的增殖和分化。

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