中国骨质疏松杂志

龙胆苦苷通过MAPK信号通路对骨质疏松大鼠骨吸收的影响

Effect of gentiopicroside on bone resorption through MAPK signaling pathway in osteoporosis rats

DOI：10.3969/j.issn.1006.7108.2022.03.008

中文关键词: 龙胆苦苷骨质疏松核因子κB受体活化因子配体丝裂原活化蛋白激酶

英文关键词:gentiopicroside osteoporosis receptor activator of nuclear factor-κB ligand mitogen-activated protein kinase

基金项目:三亚市医疗卫生科技创新项目（2015YQW09）

作者	单位
王维学* 陈希跃丰景斌罗贤红符仲秋	海南省三亚市人民医院创伤骨科，海南三亚 572000

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中文摘要:

目的基于丝裂原活化蛋白激酶（MAPK）信号通路探讨龙胆苦苷（GENT）对骨质疏松（OP）大鼠骨吸收的影响。方法 78只雌性SD大鼠随机分为假手术组（Sham组）、模型组（OP组）、雌二醇组（E2组，0.05 mg/kg）、龙胆苦苷低（GENT LD，30 mg/kg）、中（GENT MD，60 mg/kg）、高剂量组（GENT HD组，120 mg/kg），除Sham组外大鼠均采用双侧卵巢切除术构建OP大鼠模型。ELISA检测血清骨代谢指标（CBF-α1、CTX-Ⅰ、PINP、OCE2）和炎性指标（IL-6、TNF-α、IL-1β）含量；三点弯曲测试评估右股骨机械强度；Micro-CT扫描大鼠右侧股骨观察股骨远端骨微结构变化；HE染色观察骨组织形态学变化；TRAP染色分析骨表面的破骨细胞数量（N.Oc/BS）；Western Blot检测骨组织RANKL和MAPK通路相关蛋白（JNK、ERK1/2、p38 MAPK及其磷酸化蛋白）的表达。结果与Sham组相比，OP组大鼠血清E2、骨代谢相关标志物、股骨最大负荷和断裂挠度、BMD、骨小梁微结构参数BV/TV、Tb.N、Tb.Th显著降低（P＜0.05），炎症因子、Tb.Sp、N.Oc/BS、骨组织RANKL、p-JNK/JNK、p-ERK1/2/ERK1/2、p-p38/p38的表达明显升高（P＜0.05），骨皮质厚度和骨小梁数量明显减少，且骨小梁出现断裂，排列疏松；经GENT治疗后大鼠血清E2、骨代谢相关标志物、股骨最大负荷和断裂挠度、BMD、BV/TV、Tb.N、Tb.Th明显升高（P＜0.05），炎症因子、Tb.Sp、N.Oc/BS、骨组织RANKL、p-JNK/JNK、p-ERK1/2/ERK1/2、p-p38/p38的表达明显降低（P＜0.05），可见新生骨小梁，形态相对完整。结论 GENT可改善OP大鼠的骨微结构，对OP具有保护作用，其作用机制可能与抑制MAPK信号通路的激活，进而抑制破骨细胞的生成，减少骨吸收有关。

英文摘要:

Objective To investigate the effect of gentiopicroside (GENT) on bone resorption in osteoporosis (OP) rats based on mitogen-activated protein kinase (MAPK) signaling pathway. Methods Seventy-eight female SD rats were randomly divided into sham operation group (Sham group), model group (OP group), estradiol group (E2 group, 0.05 mg·kg-1), gentiopicroside low- (GENT LD, 30 mg·kg-1), medium- (GENT MD, 60 mg·kg-1), and high-dose group (GENT HD groups, 120 mg·kg-1). Except in Sham group, OP model was reproduced with bilateral ovariectomy. ELISA was used to detect the serum levels of bone metabolism indexes (CBF-α1,CTX-Ⅰ, PINP, OC, and E2), and inflammatory indexes (IL-6, TNF-α, and IL-1 β). Three-point bending test was used to evaluate the mechanical strength of the right femur. Micro CT was used to scan the right femur of rats to observe the changes of bone microstructure of the distal femur. HE staining was used to observe the changes of bone histomorphology. TRAP staining was use to analyze the number of osteoclasts on bone surface (N.Oc/BS). Western blotting was used to detect the expression of RANKL, JNK, ERK1/2, p38 MAPK, and their phosphorylated proteins. Results Compared to those in Sham group, the serum E2, bone metabolism related markers, maximum loading and fracture deflection of the femur, BMD, bone trabecular microstructure parameters BV/TV, Tb.N, and Tb.Th decreased significantly in OP group (P<0.05). Inflammatory factors, Tb.Sp, N.Oc/BS, the expression of RANKL, p-JNK/JNK, p-ERK1/2/ERK1/2, and p-p38/p38 in the bone tissue increased significantly (P<0.05). The thickness of cortical bone and the number of trabeculae reduced significantly. The trabeculae were broken and arranged loosely. After the treatment with GENT, the serum levels of E2 and bone metabolism related markers, maximum loading and fracture deflection of the femur, BMD, bone trabecular microstructure parameters BV/TV, Tb.N, and Tb.Th in E2 group and GENT MD, GENT HD groups increased significantly (P<0.05). Inflammatory factors, Tb.Sp, N.Oc/BS, the expression of RANKL, p-JNK/JNK, p-ERK1/2/ERK1/2, and p-p38/p38 in the bone tissue decreased significantly (P<0.05). The new bone trabeculae can be seen with relatively complete morphology. Conclusion GENT improves the bone microstructure of OP rats and has a protective effect from OP. Its mechanism may be related to the inhibition of the activation of MAPK signaling pathway, the formation of osteoclasts, and the reduction of bone resorption.

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