淫羊藿苷及脱水淫羊藿素对人牙髓细胞增殖和成矿能力的影响

Effects of icariin and anhydroicaritin on proliferation and mineralization of human dental pulp cells

目的观察淫羊藿苷及脱水淫羊藿素对人牙髓细胞增殖和成矿能力的影响。方法将人牙髓细胞随机分为观察A组、观察B组和对照组。观察A、B组分别加入不同浓度淫羊藿苷、脱水淫羊藿素,对照组不予处理。采用CCK-8法检测各组增殖能力(以OD450表示),Real-time PCR法检测骨钙素(OCN)和牙本质涎磷蛋白(DSPP)mRNA,碱性磷酸酶染色和茜素红染色观察各组成矿能力。结果观察A组加入0.1、1、10μmol/L淫羊藿苷后OD450分别为1.86±0.08、1.96±0.07、1.82±0.04,观察B组加入0.1、1、10μmol/L脱水淫羊藿素后OD450分别为1.90±0.07、1.93±0.09、1.88±0.02,对照组为1.76±0.10;观察A、B组均高于对照组,P均<0.05;观察A、B组比较,P均>0.05。观察A、B组OCN、DSPP mRNA表达均高于对照组,P均<0.05;观察A、B组比较,P均>0.05。在成骨诱导剂存在情况下,观察A、B组和对照组的染色结果无明显差别。结论淫羊藿苷和脱水淫羊藿素均能提高人牙髓细胞的增殖和成矿能力,但成矿能力与成骨诱导剂相比作用较微弱。

Objective To investigate the effects of icariin and anhydroicaritin on the proliferation and mineralization of human dental pulp cells( DPCs). Methods DPCs were divided into the observation groups A,B and the control group.Observation groups A and B were treated with icariin or anhydroicaritin at different concentrations,respectively; the control group was not treated. Cell proliferation was determined by CCK-8 assay( OD450 represents the absorbance value at 450 nm wavelength). The real-time PCR was used to detect the mRNA expression of mineralization-related gene osteocalcin( OCN) and dentinogensis-specific gene dentin sialophosphoprotein( DSPP). Alkaline phosphatase( ALP) histochemical staining and alizarin red staining were used to evaluate the effects of icariin and anhydroicaritin on the mineralization capacity of DPCs. Results After the cells were treated with 0. 1,1 and 10 μmol / L icariin,the OD450 values in the observation group A were 1. 86 ± 0. 08,1. 96 ± 0. 07 and 1. 82 ± 0. 04,respectively. After the cells were treated with 0. 1,1,10μmol /L anhydroicaritin,the OD450 values in the observation group B were 1. 90 ± 0. 07,1. 93 ± 0. 09 and 1. 88 ± 0. 02,respectively. Meanwhile,the OD450 value in the control group was 1. 76 ± 0. 10. The OD450 values in the observation groups A and B were higher than that in the control group( all P < 0. 05). However,there was no significant difference between observation groups A and B( all P > 0. 05). Similarly,OCN and DSPP mRNA expression in the observation groups A and B was higher than that in the control group( all P < 0. 05),and no significant difference was found between observation groups A and B( all P > 0. 05). There was no significant difference between observation groups A,B and the control group under osteoinductive condition. Conclusion Icariin and anhydroicaritin may promote proliferation and mineralization abilities of DPCs. However,compared with osteogenesis supplement,icariin and anhydroicaritin have a slight osteogenic effect on DPCs.