人参皂苷 Rg3对骨质疏松大鼠骨代谢及肠钙吸收功能的影响

Effects of Ginsenoside Rg3 on bone metabolism and intestinal calcium absorption in osteoporosis rats

摘要:

目的 探究人参皂苷Rg₃对骨质疏松（osteoporosis，OP）大鼠骨代谢及肠钙吸收功能的影响。方法 60只SPF级雌性Wistar大鼠，按照随机数字法分为空白组、模型组、阳性组、实验组，每组大鼠各15只。除空白组外，其他大鼠均采用去势（OVX法）法制备OP大鼠模型。造模成功后，空白组及模型组生理盐水灌胃[10 mL/（kg·d）]，阳性组给予阿仑膦酸钠维D3灌服(每周6.25 mg/kg），实验组给予人参皂苷Rg₃[80 mg/（kg·d）]灌胃治疗，连续干预治疗12周。检测各组股骨、胫骨骨密度变化，酶联免疫法检测大鼠血清中骨保护素（osteoclastogenesis inhibitory factor，OPG）、Ⅰ型前胶原氨基末端肽（procollagen type Ⅰ N-terminal propeptide，PINP）、抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase，TRACP）、I型胶原交联C-末端肽（Type Ⅰcollagen carboxy-terminal peptide，CTX-I）含量变化；原位末端标记法（TdT-mediated dUTP nick end labeling，TUNEL）法分析各组肠黏膜组织细胞凋亡情况；Masson法分析各组肠黏膜组织纤维病理改变；免疫组化法检测各组肠黏膜组织中维生素D膜相关快速反应结合蛋白（membrane-associated rapid response steroid-binding，1，25-D3-MARRS）蛋白表达；蛋白免疫印迹法及RT-PCR法检测各组Jagged1、Notch1、Hes1蛋白及mRNA表达水平。结果 大鼠造模后股骨、胫骨骨密度明显下降，血清中OPG、PINP、TRACP及CTX-I含量明显变化（P<0.05）；肠黏膜组织病理发生明显改变，组织纤维化增强，细胞凋亡程度增加；肠黏膜组织1，25-D3-MARRS蛋白表达降低（P<0.05）。治疗后与模型组对比，阳性组及实验组大鼠股骨、胫骨骨密度明显升高，血清中OPG、PINP、TRACP及CTX-I含量明显改善（P<0.05）；肠黏膜组织病理发生明显改善，纤维化降低，细胞凋亡程度降低；肠黏膜组织中Jagged1、Notch1、Hes1蛋白及mRNA表达明显改善（P<0.05）。结论 人参皂苷 Rg₃能够提高OP大鼠骨密度，减轻肠黏膜组织细胞凋亡及组织纤维化，增加肠黏膜1，25D3-MARRS蛋白表达，改善肠道钙吸收。

Abstract:

Objective To explore the effect of Ginsenoside Rg3 on bone metabolism and intestinal calcium absorption in osteoporosis（OP） rats.Methods 60 SPF grade female Wistar rats were randomly divided into blank group，model group，positive group，and experimental group，with 15 rats in each group. Except for the blank group，all other rats were used to prepare OP rat models using the OVX method. After successful modeling，the blank group and the model group were given normal saline by gavage[10 mL/（kg·d）]，the positive group was given alendronic acid sodium and vitamin D3 by gavage[6.25 mg/（kg·w）]，and the experimental group was given ginsenoside Rg3[80 mg/（kg·d）] by gavage for 12 consecutive weeks. Bone mineral density changes of femur and tibia in each group were detected. Serum osteoprotegerin（OPG），procollagen type I N-terminal peptide（PINP），tartrate resistant acid phosphatase（TRACP） Changes in the content of Type I collagen cross-linked C-terminal peptide（CTX-I）；TUNEL method was used to analyze the apoptosis of intestinal mucosal tissue cells in each group；Masson's method was used to analyze the pathological changes of intestinal mucosal tissue fibers in each group；Immunohistochemical method was used to detect the protein expression of vitamin D membrane associated rapid response steroid binding（1，25-D3-MARRS） in intestinal mucosa tissues of each group；Protein immunoblotting and RT-PCR were used to detect the expression levels of Jagged1，Notch1，Hes1 protein and mRNA in each group.Results After modeling，the bone density of the femur and tibia in rats decreased significantly，and the levels of OPG，PINP，TRACP，and CTX-I in serum showed significant changes（P<0.05）；Significant pathological changes occur in the intestinal mucosa tissue，with increased fibrosis and increased degree of cell apoptosis；The expression of 1，25-D3-MARRS protein in intestinal mucosal tissue decreased（P<0.05）. After treatment，compared with the model group，the bone density of the femur and tibia of the positive and experimental groups were significantly increased，and the serum levels of OPG，PINP，TRACP，and CTX-I were significantly improved（P<0.05）；The pathological changes of intestinal mucosa tissue have significantly improved，with reduced fibrosis and reduced degree of cell apoptosis；The expression of Jagged1，Notch1，Hes1 proteins and mRNA in intestinal mucosal tissue was significantly improved（P<0.05）.Conclusion Ginsenoside Rg3 can increase bone density of OP rats，reduce apoptosis and fibrosis of intestinal mucosa，increase the expression of 1，25D3-MARRS protein in intestinal mucosa，and improve intestinal calcium absorption.