摘要
【目的】观察滋肾降糖丸对糖尿病大鼠骨代谢指标、骨密度及骨组织形态的影响及其对Wnt/β-连环蛋白(β-catenin)通路的调控作用,探讨其对糖尿病介导的骨量减少及骨质疏松形成的防治机制。【方法】采用腹腔注射链脲佐菌素(STZ)法建立糖尿病大鼠模型。将造模成功的大鼠随机分为模型组、中药组、西药组,另设正常组,每组10只。中药组大鼠给予滋肾降糖丸浓缩液(剂量为5.85 g·kg^(-1)·d^(-1))灌胃,西药组大鼠给予雷奈酸锶浓缩液(剂量为0.9 g·kg^(-1)·d^(-1))灌胃,正常组与模型组大鼠给予等体积生理盐水灌胃。干预3个月后,观察各组大鼠生化指标的变化;采用苏木素—伊红(HE)染色法观察各组大鼠骨组织形态变化;采用实时定量PCR(qPCR)方法检测骨组织Wnt3α、低密度脂蛋白受体相关蛋白5(LRP-5)、β-catenin、Runt相关转录因子2(RUNX-2)、Dickkopf相关蛋白1(DKK-1)、硬化蛋白(SOST)基因表达水平。【结果】(1)与模型组比较,中药组大鼠空腹胰岛素(Fins)、血钙(BC)、血磷(BP)、骨特异性碱性磷酸酶(B-ALP)、骨钙素(OC)、骨密度(BMD)水平均升高(P<0.01),空腹血糖(FBG)、β-I型胶原羧基端肽(β-CTX)、1型前胶原酶N端肽(P1NP)、骨保护素(OPG)水平均明显降低(P<0.01),骨组织Wnt3α、LRP-5、β-catenin、RUNX-2基因表达量升高(P <0.01),而DKK-1及SOST基因表达量无显著性差异(P> 0.05)。(2)中药组骨组织结构紊乱较模型组明显减轻,基本接近正常。【结论】滋肾降糖丸可能通过上调Wnt/β-catenin信号通路中重要分子Wnt3α、LRP-5、β-catenin、RUNX-2的基因表达,促进成骨细胞的分化,抑制骨破坏,改善骨组织形态,增加骨密度,调节骨代谢紊乱,进而发挥对糖尿病大鼠骨量减少和骨质疏松形成的防治作用。
Objective To observe the influence of Zishen Jiangtang Pills on bone metabolism indexes, bone mineral density and histomorphological characteristics of bone tissues in diabetic rats,and to explore its regulatoryeffect on Wnt/β-catenin signaling pathway,thus to explore the preventive and therapeutic mechanism of Zishen Jiangtang Pills for diabetes-induced bone mass loss and formation of osteoporosis. Methods The rat model ofdiabetes was established by intraperitoneal injection of streptozotocin（STZ）. The modeled rats were randomlydivided into model group, Chinese medicine group, and western medicine group, 10 rats in each group.Additionally, 10 healthy rats were served as the normal group. Chinese medicine group was given intragastricadministration of Zishen Jiangtang Pills（at the dosage of 5.85 g·kg-1·d-1）,and western medicine group was givenintragastric administration of Strontium Ranelate concentrated solution（at the dosage of 0.9 g·kg-1·d-1）,and thenormal group and the model group were given intragastric administration of the same volume of normal saline water.After 3-month medication, the changes of biochemical indexes in various groups were observed, thehistomorphological features of bone tissues in various groups were examined by hematoxylin and eosin（HE）staining method, and the gene expression levels of Wnt3, low-density lipoprotein receptor-related protein 5（LRP-5）,β-catenin,Runt-related transcription factor 2（RUNX-2）,Dickkopf-1（DKK-1）and sclerostin（SOST）were detected by real-time quantitative PCR（qPCR）. Results（1）Compared with the model group,thelevels of fasting insulin（Fins）,blood calcium（BC）,blood phosphorus（BP）,bone alkaline phosphatase（B-ALP）,osteocalcin（OC）,bone mineral density（BMD）were increased（P〈0.01）,and the levels of fasting bloodglucose（FBG）and beta isomer of the C-terminal telopeptide of type Ⅰ collagen（β-CTX）,procollagen 1 amino-terminal propeptide（P1 NP）and osteoprotegerin（OPG）were significantly decreased（P〈0.01）,and the geneexpression levels of Wnt3α、LRP-5、β-catenin、RUNX-2 in rat bone tissues were increased（P 〈 0.01）,whileDKK-1 and SOST had no obvious changes（P〉0.05）in Chinese medicine group.（2）The disorder of rat bonetissue structure in Chinese medicine group was obviously improved as compared with that in the model group,nearly close to the normal level. Conclusion Zishen Jiangtang Pills play a role in preventing and treating bone massloss and formation of osteoporosis in diabetes rats through up-regulating the gene expression of important moleculesWnt3α、LRP-5、β-catenin and RUNX-2 in Wnt/β-catenin signaling pathway,promoting the formation anddifferentiation of osteoblast, inhibiting bone destruction, improving the histomorphology of bone tissues,increasing bone mineral density,and regulating bone metabolism disorder.
作者
赵恒侠
周道成
陈叶
ZHAO Heng-Xia;ZHOU Dao-Cheng;CHEN Ye(Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 518033 Guangdong,China)
出处
《广州中医药大学学报》
CAS
2018年第6期1051-1057,共7页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
深圳市科技计划项目(编号:JCYJ20150401163841053)
深圳市三名工程项目(编号:SZSM201512043)
国家中医药管理局全国名老中医药专家传承工作室建设项目(2012年)
