摘要
目的:探讨补肾健脾方药对尾部悬吊大鼠骨髓间充质干细胞成骨诱导分化后GSK-3β、β-catenin表达的影响。方法:随机将30只大鼠分为正常组、悬吊组、补肾健脾组,悬吊组、补肾健脾组采用头低位-30°尾部悬吊法模拟失重,补肾健脾组给予补肾健脾方灌服,其他两组给予等量0.9%氯化钠溶液。灌胃第22天取各组大鼠双侧胫骨行HE染色进行骨组织形态学观察,取双侧股骨和胫骨分离培养骨髓间充质干细胞(BMSCs)进行培养,于诱导第15天,取出96孔板,采用p-NPP法检测BMSCs ALP活性,免疫组织化学染色及Western blot法检测BMSCs GSK-3β、β-catenin的表达情况。结果:与正常组比较,悬吊组BMSCs ALP活性明显降低(P<0.01);与悬吊组比较,补肾健脾组BMSCs ALP活性升高(P<0.05)。与正常组比较,悬吊组BMSCs核中GSK-3β表达较强、蛋白表达水平显著升高(P<0.01),β-catenin表达水平显著降低(P<0.01);与悬吊组比较,补肾健脾组BMSCs核中GSK-3β表达降低、蛋白表达水平降低(P<0.05),β-catenin表达显著升高(P<0.05)。结论:补肾健脾方对尾部悬吊大鼠的骨丢失具有防治作用,其作用机制可能与促进BMSCs成骨诱导分化后β-catenin的表达、抑制GSK-3β的表达,激活Wnt通路有关,通过Wnt信号通路的激活,以促进骨形成。
Objective：To observe the effects of Bushen Jianpi Formula on the expression of GSK-3β,β-catenin in bone marrow stem cells（BMSCs） of tail suspension rats.Methods：Thirty rats were randomly divided into control（C） group,tail suspended（S） group,Bushen Jianpi Formula（TKIS） group.Rats in S and TKIS groups were simulated weightlessness by head down tilt-30° tail suspension.TKIS groups were intragastric administrated with Bushen Jianpi Formula and other groups were administrated with 0.9% normal saline by gavage.After 22 days,the morphology of bone tissue was observed by HE staining of bilateral tibia,and the BMSCs from the bilateral femur and tibia of each group were isolated and cultured.After 15 days,the activity of BMSCs ALP was detected by using p-NPP method in the 96-well plates,and the expression of BMSCs GSK-3β,β-catenin were detected by immunohistochemistry and Western blot.Results：Compared with the C group,the BMSCs ALP activity of S group was significantly decreased（P〈0.01）;compared with S group,the BMSCs ALP activity of TKIS group was increased（P〈0.05）.Compared with the C group,the expression of GSK-3β in BMSCs was higher,while protein levels were significantly increased（P〈0.01） and β-catenin expression was significantly decreased（P〈0.01）.Compared with the S group,the expression of GSK-3β and protein in BMSCs were decreased（P〈0.05）,while β-catenin expression was significantly increased（P〈0.05）.Bushen Jianpi Formula had prevention and treatment effect on the bone loss of tail suspension rats.Its mechanism might be related to promote the expression of β-catenin after the osteoinductive culture of the BMSCs,inhibit the expression of GSK-3β,activate the Wnt signaling pathway,and promote bone formation by activating the Wnt signaling pathway.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2016年第9期3745-3748,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81001493)
辽宁省高等学校杰出青年学者成长计划(No.LJQ2014103)~~
关键词
模拟失重
骨丢失
骨髓间充质干细胞
Wnt信号通路
补肾健脾
Simulated weightlessness
Bone loss
BMSCs
Wnt signaling pathway
Invigorating kidney for strengthening spleen
