刺老苞根皮含药血清对原代成骨细胞Wnt/βcatenin信号通路的影响

依香叫　李金诚　王松月　燕梦云　崔箭　裴凌鹏

[摘要]该文研究了不同浓度刺老苞根皮含药血清对原代成骨细胞Wnt/βcatenin信号通路中βcatenin，Wnt1，Frizzed2，TCF和Axin表达的影响。经SPF级健康雌性SD大鼠（80只）给予蒸馏水、刺老苞根皮水煎剂高、中、低3种剂量，连续灌胃7 d，制备刺老苞根皮含药血清。体外培养原代成骨细胞（OB）并鉴定，取第3代成骨细胞，培养48 h后用各组含药血清干预培养10 d，采用茜素红染色钙化结节并计数；干预培养48 h后收集细胞，分别采用Realtime PCR（RTPCR）和Western blot 法测定βcatenin，Wnt1，Frizzed2，TCF和Axin的表達情况。结果显示，经鉴定体外培养细胞为原代成骨细胞；在干预培养后，与模型组相比，各剂量组可促进原代成骨细胞的矿化能力，且上调βcatenin，Wnt1，Frizzed2，TCF mRNA和蛋白的表达，下调Axin mRNA和蛋白的表达。结果发现刺老苞根皮含药血清可通过调节原代成骨细胞Wnt/βcatenin信号通路中βcatenin，Wnt1，Frizzed2，TCF和Axin表达，增强成骨细胞的分化和增殖。

[关键词]刺老苞根皮； 原代成骨细胞； Wnt/βcatenin； 信号通路

[Abstract]This paper was aimed to investigate the effect of Aralia echinocaulis containing serum on expression of βcatenin， Wnt1， Frizzed2， TCF and Axin in Wnt/βcatenin signaling pathway of primary osteoblasts SD healthy female rats （n=80） were used to make A echinocaulis containing serum by gastric perfusion for seven days with distilled water， A echinocaulis decoction high dosage， middle dosage， and low dosage In vitro， primary osteoblasts were cultured and identified The third generation primary osteoblasts were taken and cultured for 48 h， then cells were treated with the different drug serums for 10 days and calcified nodules were counted by alizarin red staining The cells were collected after treatment for 48 h and the expression levels of βcatenin， Wnt1， Frizzled2， TCF and Axin were detected by Realtime PCR and Western blot The results suggested that the in vitro cells were primary osteoblasts； and after treatment， various doses groups could promote the mineralization ability of primary osteoblasts， upregulate the mRNA and protein expression levels of βcatenin， Wnt1， Frizzled2， and TCF， and downregulate the mRNA and protein expression levels of Axin These findings indicated that A echinocaulis containing serum can enhance the differentiation and proliferation of osteoblasts by regulating the expression levels of βcatenin， Wnt1， Frizzled2， TCF and Axin in Wnt/βcatenin signaling pathway of primary osteoblasts

[Key words]Aralia echinocaulis； primary osteoblasts； Wnt/βcatenin； signaling pathway

近年来，民族药物资源的开发和利用倍受国内外研究学者的关注和青睐，传统土家族医药——刺老苞根皮为五加科植物楤木Aralia chinensis L或棘茎楤木A echinocaulis HandMazz 的根皮或茎皮，分布于我国西南、华南及华东山区，包括云南东南部及西双版纳地区、贵州黔南州及黔东南州、广西壮族自治区、海南省五指山周围地区、广东省山区、福建省中南部地区、江西省南端、台湾省等地山区，资源非常丰富[12]。该药材片状或槽状，气微香，嚼之带黏液性，味辛，性平。归肝、肾经，功能滋阴健肾，祛风湿，壮筋骨，散瘀血，消肿毒。可用于风湿痹痛、跌打损伤、骨折等治疗[34]。……