Song Peng, Yao Juan, Ma Hui-Ping, Ge Bao-Feng, Chen Ke-Ming, Guo Xiao-Yu, Lü Xiang
Institute of Orthopaedics, Lanzhou General Hospital of PLA, Lanzhou 730050, China.
Yao Xue Xue Bao. 2012 Jul;47(7):890-6.
This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.
本研究旨在比较山柰酚和脱水淫羊藿素对大鼠成骨细胞(ROB)体外生物矿化的影响。无菌解剖新生SD大鼠的颅骨,通过酶消化获得成骨细胞,并在含有10%胎牛血清的MEM中培养。每三天更换一次培养基,当细胞覆盖培养皿的90%时进行连续传代培养。在成骨分化条件下,分别加入终浓度为1×10⁻⁴、1×10⁻⁵、1×10⁻⁶和1×10⁻⁷mol·L⁻¹的山柰酚和脱水淫羊藿素。采用MTT法检测细胞增殖情况,并在成骨诱导培养第9天通过碱性磷酸酶(ALP)活性检测最佳浓度。比较山柰酚、脱水淫羊藿素和最佳浓度对照组的成骨指标。结果表明,浓度为1×10⁻⁵mol·L⁻¹的脱水淫羊藿素显著促进了ALP活性、钙含量和骨钙素含量,增加了CFU-F(ALP)数量和矿化结节,提高了成骨分化关键基因BMP-2、OSX和Runx-2的mRNA水平,并提高了I型胶原蛋白的蛋白含量。然而,山柰酚组未显著表现出促进ROB成骨分化的能力。山柰酚和脱水淫羊藿素对ROB成骨分化的差异是由淫羊藿苷C-8位的异戊烯基引起的。