香砂六君子汤通过Notch信号通路调控骨组织成骨/成脂平衡防治绝经后骨质疏松症实验研究  

Experimental Study on Prevention and Treatment of Postmenopausal Osteoporosis by Regulating Bone Tissue Osteogenesis/Lipogenesis Balance through Notch Signaling Pathway with Xiangsha Liujunzi Decoction(香砂六君子汤)

作　　者：乔隆 关雪峰 乔野 于冬冬[2] 齐鹏坤[1,2] 李泽 齐文诚[2] QIAO Long;GUAN Xuefeng;QIAO Ye;YU Dongdong;QI Pengkun;LI Ze;QI Wencheng(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Shenyang Pharmaceutical University,Shenyang 117004,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032 [3]沈阳药科大学,辽宁沈阳117004

出　　处：《中华中医药学刊》2023年第10期168-172,I0032-I0037,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(8170933);辽宁省自然科学基金指导项目博士启动资金项目(2022-BS-202);辽宁省教育厅科学研究经费项目(L202032)。

摘　　要：目的 探讨香砂六君子汤防治去卵巢骨质疏松模型大鼠(prevention and treatment of ovariectomy osteoporosis model, OVX)大鼠的作用机制。方法 取SD雌性大鼠60只,随机分为空白组、模型组、仙灵骨葆胶囊组、香砂六君子汤(低剂量、中剂量、高剂量)给药组,除空白组外其余各组采用卵巢去势手术诱导绝经后大鼠骨质疏松模型,灌胃给药8周后取材,留取各组大鼠血清以及股骨。酶联免疫吸附试验(Enzyme linked immunosorbent assay, ELISA)法检测骨代谢指标碱性磷酸酶(alkaline phosphatase, ALP)、Ⅰ型前胶原氨基端延长肽(typeⅠprocollagen N-terminal propeptide, P1NP)、Ⅰ型胶原羧基端肽β(typeⅠcollagen carboxyterminal peptide β-specific sequence, β-CTX);生物力学检测骨强度,Micro-CT观察骨组织形态及骨密度;苏木精-伊红染色法(hematoxylin-eosin staining, HE)染色检测骨结构以及骨髓脂肪含量,免疫组织化学分析成骨蛋白Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)、成脂蛋白过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptors, PPARγ)、通路蛋白Notch1的表达情况。结果 香砂六君子汤可以提高大鼠股骨最大荷载力,提高大鼠股骨的骨小梁面积和骨小梁数目,促进血清ALP、P1NP表达,抑制β-CTX表达,抑制骨髓腔内脂肪粒含量,免疫组化结果发现香砂六君子汤可促进RUNX2、Notch1蛋白的表达,抑制PPARγ蛋白的表达。结论 香砂六君子汤能够促进骨形成、增加骨量、提高骨密度、改善骨结构、增强骨强度、降低骨髓腔脂肪含量,防治绝经后骨质疏松症(postmenopausal osteoporosis, PMOP)。香砂六君子汤在防治PMOP的过程中,可通过激活Notch通路来调RUNX2和下调PPAPγ的表达,促进成骨,从而治疗骨质疏松症。Objective To investigate the action mechanism of Xiangsha Liujunzi Decoction(香砂六君子汤)in the prevention and treatment of ovariectomy osteoporosis model(OVX)rats.s Methods Sixty female rats with SD were randomly divided into blank group,model group,Xianling Gubao Tablets(仙灵骨葆片)administration group,Xiangsha Liujunzi Decoction(low,medium and high dose)administration groups.Except the normal group,the remaining groups used ovarian castration surgery to induce postmenopausal rat osteoporosis model.After 8 weeks of intragastric administration,samples were taken and serum and femur samples were collected from each group of rats.ELISA method was used to detect bone metabolism indicators[alkaline phospha-tase(ALP),typeⅠprocollagen N-terminal propeptide(P1NP)and TypeⅠcollagen carboxyterminal peptideβ-specific se-quence(β-CTX)].Biomechanical method was used to detect the bone strength.Micro-CT observation was for bone tissue morphology and bone density.HE staining detected bone structure and bone marrow fat content,and immunohistochemistry ana-lyzed the expressions of osteogenesis protein RUNX 2,lipogenic protein PPARγand pathway protein Notch1.s Results Xiangsha Liujunzi Decoction could increase the maximal load capacity of the femur,increase the area of bone trabeculae and the number of trabecular bones of femur,promote serum ALP and P1NP expressions,inhibitβ-CTX expression,inhibit the contents of fat parti-cles in the bone marrow cavity of the rats.And the immunohistochemistry method has found that Xiangsha Liujunzi Decoction could promote the expressions of RUNX2 and Notch1 proteins and inhibit the expression of PPARγprotein.n Conclusion Xiangsha Liujunzi Decoction can promote bone formation,increase bone mass,improve bone density and structure,enhance bone strength,reduce bone marrow cavity fat content and prevent postmenopausal osteoporosis(PMOP).In the process of preventing PMOP,it also can up-regulate the expression of RUNX2 and down-regulate PPAPγby activating the Notch pathway so as t

关 键 词：骨质疏松症 NOTCH通路 香砂六君子汤 

分 类 号：R285.5[医药卫生—中药学]