补肾健脾活血方对UMR106细胞成骨分化及ERα的影响

Effect of Kidney-tonifying Spleen-invigorating Blood-activating Prescription on Osteogenic Differentiation and ERαof UMR106

目的:探讨补肾健脾活血方对过表达分泌型卷曲相关蛋白1(secreted frizzled related protein 1,SFRP1)、沉默SFRP1的UMR106细胞成骨分化及雌激素受体α(estrogen receptorα,ERα)的影响。方法:通过构建SFRP1过表达及沉默重组腺病毒载体,并转染大鼠类成骨细胞系UMR106细胞,初步分为空载腺病毒组、过表达SFRP1组、沉默SFRP1组,并根据含药血清和生理盐水(空白)血清干预的不同分为6组,观察6组细胞的碱性磷酸酶(alkaline phosphatase,ALP)活性及细胞ERα蛋白表达情况。结果:含药血清干预的空载腺病毒组、SFRP1沉默组及SFRP1过表达组72 h后UMR106细胞ALP活性和ERα蛋白表达均高于空白血清干预的空载腺病毒组、SFRP1沉默组及SFRP1过表达组(P<0.05);空白血清+SFRP1沉默组的UMR106细胞ALP活性及ERα蛋白表达高于空白血清+空载腺病毒组(P<0.05),而空白血清+SFRP1过表达组的UMR106细胞ALP活性及ERα蛋白表达低于空白血清+空载腺病毒组(P<0.05)。结论:过表达SFRP1可以抑制UMR106细胞成骨分化,并下调ERα蛋白表达;沉默SFRP1和补肾健脾活血方均可促进UMR106细胞成骨分化,并上调ERα蛋白表达,且两者共同干预时作用更显著,说明补肾健脾活血方能够抑制SFRP1表达,而SFRP1并不是补肾健脾活血方调节成骨细胞代谢,提高成骨分化活性和促进ERα蛋白表达的唯一靶点,可能存在其他靶点共同促进调节成骨细胞代谢。

Objective:To explore the influence of kidney-tonifying spleen-invigorating blood-activating prescription on osteogenic differentiation and estrogen receptorα(ERα)of UMR106 cells via SFRP1 overexpression or gene knockdown.Methods:After constructing recombinant adenoviral vectors for SFRP1 overexpression or gene knockdown,rat osteoblast-like UMR106 cells,transfected with the virus,were primarily divided into empty adenovirus group,overexpression SFRP1 group and silencing SFRP1 group,and they were allocated to six groups according to the intervention with the medicated serum and physiological saline(blank)serum,to observe the activity of ALP and the expressions of cellular ERαin six groups.Results:After intervening with medicated serum for 72 hours,the activity of ALP and the expressions of cellular ERαin empty adenovirus group,silencing SFRP1 group and overexpression SFRP1 group were higher than these of empty adenovirus group,silencing SFRP1 group and overexpression SFRP1 group intervened with blank serum(P<0.05);ALP activity and the expressions of ERαprotein in blank serum+silencing SFRP1 group were higher than these of blank serum+empty adenovirus group(P<0.05),while ALP activity and the expressions of ERαprotein in UMR106 cells of blank serum+overexpression SFRP1 group were lower than these in blank serum+empty adenovirus group(P<0.05).Conclusion:Overexpression of SFRP1 could inhibit osteogenic differentiation of UMR106 cells and downregulate the expressions of ERαprotein;Silencing of SFRP1 and kidney-tonifying spleeninvigorating blood-activating prescription could promote osteogenic differentiation of UMR106 cells,up regulate the expressions of ERαprotein,and the effects are more evident when the two acted together,demonstrating that the formula could inhibit SFRP1 expression,but SFRP1 is not the only target in the process of the formula regulating the metabolism of osteoblast,improving the activity of osteogenic differentiation and ERαprotein expressions,and the other targets may exist which could promote the regulation of osteoblast metabolism.