目的 研究龍牙楤木皂苷Ⅳ（Tarasaponin Ⅳ）對(duì)原代成骨細(xì)胞的分化及礦化的影響。方法 通過(guò)消化SD乳鼠顱蓋骨，獲得原代前成骨細(xì)胞，體外誘導(dǎo)培養(yǎng)并鑒定；采用MTT比色法檢測(cè)Tarasaponin Ⅳ（10、20、30、40、50、60、70、80、90、100 μg/mL）作用于前成骨細(xì)胞的適宜質(zhì)量濃度范圍；用低、中、高濃度（0.5、5、50 μg/mL） Tarasaponin Ⅳ對(duì)成骨細(xì)胞進(jìn)行干預(yù)，培養(yǎng)4 d后，試劑盒法進(jìn)行細(xì)胞上清中堿性磷酸酶（ALP）活性測(cè)定；培養(yǎng)14 d后，按茜素紅S染色試劑說(shuō)明對(duì)細(xì)胞進(jìn)行鈣化結(jié)節(jié)染色。結(jié)果 與對(duì)照組比較，Tarasaponin Ⅳ各濃度組均能明顯提高前成骨細(xì)胞活性（P<0.05、0.01），0~60 μg/mL內(nèi)細(xì)胞生存率呈遞增趨勢(shì)。與對(duì)照組比較，Tarasaponin Ⅳ低、中、高濃度組細(xì)胞上清ALP活性均顯著下降（P<0.01），中、高濃度組細(xì)胞礦化結(jié)節(jié)數(shù)目顯著增多（P<0.01），且均呈劑量相關(guān)性。結(jié)論 Tarasaponin Ⅳ可以促進(jìn)前成骨細(xì)胞生長(zhǎng)，增強(qiáng)成骨細(xì)胞礦化過(guò)程。

Objectives To study the influence of Tarasaponin Ⅳ on the differentiation and mineralization of primary osteoblasts.Methods The craniums of male SD neonatal rats were digested to obtain primary pre-osteoblasts,and cells were then cultured in vitro and identified.The suitable concentration range of Tarasaponin Ⅳ(10,20,30,40,50,60,70,80,90,100 μg/mL) was determined by MTT colorimetry.The osteoblasts were respectively cultured with Tarasaponin Ⅳ(0.5,5,50 μg/mL).Alkaline phosphatase (ALP) activity in cell supernatant was determined by kit method after 4 d of culture,and calcified nodules were stained by alizarin red S staining after 14 d of culture.Results Compared with the control group,all concentration groups of Tarasaponin Ⅳ could significantly increase the activity of osteoblasts (P< 0.05,0.01),and the cell survival rate showed an increasing trend in the concentration range of 0-60 μg/mL.Compared with the control group,the ALP activities in the supernatant of Tarasaponin Ⅳ low,medium,and high dose groups were decreased significantly (P< 0.01),and the number of mineralized nodules in the medium and high concentration groups was increased significantly (P< 0.01),in a dose-dependent manner.Conclusion Tarasaponin Ⅳ can promote the proliferation of pre-osteoblast and enhance the mineralization process of osteoblasts.

國(guó)家自然科學(xué)基金資助項(xiàng)目（81673769、81473451）；中央民族大學(xué)碩士研究生自主科研項(xiàng)目（182150）