ERK/Smad信号通路影响固本增骨方对去势大鼠骨代谢的研究

Effect of Guben Zenggu prescription on bone metabolism in ovariectomized rats based on the ERK/Smad signaling pathway

目的由ERK/Smad信号通路探讨固本增骨方对骨质疏松模型大鼠的作用机制,揭示其防治骨质疏松症的药理学效应。方法将SPF级Wistar大鼠,分为固本增骨方低、中、高剂量组,雌二醇组、假手术组及模型组,每组10只,固本增骨方低、中、高剂量组分别以固本增骨方6.1 g/(kg·d)、12.2 g/(kg·d)、24.4 g/(kg·d)的剂量进行灌胃,雌二醇组给予0.09 mg/(kg·d)的戊酸雌二醇灌胃,假手术组和模型组给予等体积生理盐水灌胃。末次灌胃后心脏采血并取材,通过ELISA检测血清BALP、TRACP-5b水平,DXA检测大鼠右侧股骨近端骨密度,并进行HE染色。运用RT-PCR检测ERK-1、ERK-2、Smad4 mRNA表达,Western Blot检测ERK、p-ERK及Smad4蛋白表达。结果经固本增骨方干预后,与模型组比较,中、高剂量可提高骨密度,骨小梁形态明显改善,血清BALPBGP水平,ERK-1、ERK-2、Smad4 mRNA显著增加,ERK、p-ERK及Smad4蛋白表达显著上升(P<0.05),血清TRACP水平显著降低(P<0.05)。结论固本增骨方可改善绝经后骨质疏松模型大鼠的骨代谢,提高骨密度,可能与激活ERK/Smad信号通路促进成骨分化有关。

Objective To explore the mechanism of action of Guben Zenggu prescription on osteoporosis model rats based on the ERK/Smad signaling pathway,and to reveal its pharmacological effects on preventing and treating osteoporosis.Methods SPF grade Wistar rats were divided into high,medium,and low dose groups of Guben Zenggu prescription,estradiol group,sham operation group,and model group,with 10 rats each.Low,medium,and high doses of Guben Zenggu prescription were gavaged with 6.1 g/(kg·d),12.2 g/(kg·d),and 24.4 g/(kg·d).Sham operation group and model group were given equal volume normal saline irrigation.Blood was collected from the heart after the last intragastric administration,the serum BALP and TRACP-5b levels were detected by ELISA.Bone mineral density of the proximal femur of the rats was measured by DXA,and HE staining was performed.RT-PCR was used to detect the mRNA expression of ERK-1,ERK-2,and Smad4,while Western blot was used to detect the protein expression of ERK,p-ERK,and Smad4.Results The intervention of Guben Zenggu prescription,compared with the model group,medium and high doses can increase bone density,significantly improve bone trabecular morphology,significantly increase serum BALP and BGP levels,ERK-1,ERK-2,and Smad4 mRNA,significantly increase the expression of ERK,p-ERK,and Smad4 proteins(P<0.05),and significantly decrease serum TRACP levels(P<0.05).Conclusion Guben Zenggu prescription can improve bone metabolism and increase bone density in postmenopausal osteoporosis model rats,which may be related to activating the ERK/Smad signaling pathway to promote osteogenic differentiation.