左归丸对同种异性骨髓移植小鼠肝组织Wnt信号通路的影响

Effects of Zuoguiwan on the Wnt signaling pathway in female mice with male mice bone marrow transplant

目的:探讨左归丸对同种异性骨髓移植小鼠肝组织Wnt信号通路的影响。方法:建立同种异性骨髓移植小鼠模型,随机分为补肾组和对照组,每组40只,分别给予左归丸汤剂和生理盐水灌胃,给药6个月后处死小鼠,取小鼠肝组织,采用基因芯片技术检测同种异性骨髓移植小鼠肝组织基因表达谱,采用Western Blot检测β-catenin蛋白表达。结果:补肾组相对于对照组小鼠肝组织Wnt信号通路中上调表达的基因有Wnt1、分泌型Frz相关蛋白(Secreted frizzled-related sequence protein,sfrp)1、sfrp5,下调表达的基因有Gsk3b、Wnt10a、Rac1、Mapk9、Prkcc、Frat1、Nfatc4、Csnk2b、Ppard等。补肾组比对照组小鼠肝组织中β-catenin蛋白表达明显增强。结论:左归丸可通过激活Wnt信号通路经典途径,游离β-catenin蛋白积累并进入细胞核激活下游靶基因,这可能是"补肾生髓成肝"调控肝再生的重要分子机制之一。

Objective： This research is to observe the effects of Zuoguiwan on the Wnt signaling pathway in female mice with male mice bone marrow transplant. Methods： We established the animal model that the recipient mice were injected via the tail vein with the bonemarrow cell suspension （0. 2ml, about 2 × 10^6 cells） of homogenic donor mice after exposed to irradiation with 9. 0Gy 60Co γ-rays. The recipient mice were randomly divided into nourish-kidney group and control group, and were respectively treated with Zuoguiwan and physiological saline. After treated 6 months, the mice were sacrificed to detect the gene expression profile with gene chip, and to observe the expression of β-catenin protein with Western Blot. Results： We found that contracted with control group, 3 genes （Wnt1, sFRP1 and sFRP5 ） of nourish-kidney group were up regulated, and 9 genes （GSK3b, Wnt10a, Racl, MAPK9, Prkcc, Fratl, Nfatc4, CSNK2b and Ppard） of nourish-kidney group were down regulated. Therefore, the expression of β-catenin protein in nourish-kidney group was apparently stronger than which in control group. Conclusion： Nourish kidney could activate classical way of Wnt signal pathway, and accumulated β-catenin protein transferred into nuclei to activate target genes maybe one of mechanisms that nourish kidney to promote bone marrow forming liver cell.