摘要
目的:探究益肾补骨汤对骨质疏松症大鼠骨形成及一氧化氮-环磷酸鸟苷(NO-cGMP)信号通路的影响。方法:采用摘除双侧卵巢法建立绝经后骨质疏松症(PMOP)SD大鼠模型,将大鼠按照随机数字表法分为假手术组(sham组)、PMOP大鼠模型组(PMOP组),益肾补骨汤L组、M组和H组(分别给予益肾补骨汤2、4和8 mL/kg灌胃干预),以及阳性对照组(给予戊酸雌二醇灌胃干预)。比较各组大鼠术前和术后体重,观察各组大鼠一般状态,采用酶联免疫吸附试验检测血清中总碱性磷酸酶(TALP)、血清抗酒石酸酸性磷酸酶(TRAP)含量,双能X线检测大鼠骨密度,苏木精-伊红染色检测大鼠股骨组织形态学变化,分别用分光光度计法和放射免疫法检测大鼠股骨组织中NO、cGMP含量;采用蛋白质印迹法检测股骨组织中Wnt3a、β-catenin蛋白表达。结果:与sham组相比,PMOP组大鼠体重明显降低(P<0.05);与PMOP组相比,益肾补骨汤L组、M组和H组大鼠体重明显升高(P<0.05),差异均有统计学意义。PMOP组大鼠血清TALP含量和股骨BMD低于sham组,血清TRAP含量高于sham组(P<0.05);益肾补骨汤L组、M组和H组血清TALP含量和股骨BMD高于PMOP组,血清TRAP含量低于PMOP组(P<0.05),且具有浓度依赖性,差异均有统计学意义。益肾补骨汤H组和阳性对照组大鼠血清TALP、TRAP含量和股骨BMD的差异无统计学意义(P>0.05)。与sham组比较,PMOP组大鼠骨小梁间隙变大不规则,粗细不均匀,骨小梁表型更薄;与PMOP组比较,益肾补骨汤L组、M组、H组以及阳性对照组大鼠骨小梁间隙均不同程度变小,且骨小梁粗细均匀且逐渐规则。与sham组比较,PMOP组股骨组织中NO、cGMP含量明显升高(P<0.05);益肾补骨汤L组、M组和H组大鼠股骨组织中NO、cGMP含量低于PMOP组(P<0.05),差异均有统计学意义。与sham组比较,PMOP组大鼠股骨组织中Wnt3a、β-catenin蛋白表达明显降低(P<0.05);益肾补骨汤L组、M组和H组大鼠股骨组织中Wnt3a、β-catenin蛋白表达明显高于PMOP组(P<0.05),且呈剂量依赖性,差异均有统计学意义。结论:益肾补骨汤可促进PMOP大鼠骨形成,可能是通过抑制NO-cGMP信号通路抑制骨吸收。
OBJECTIVE:To probe into the effects of Yishen Bugu decoction on bone formation and nitric oxide-cyclic guanosine monophosphate(NO-cGMP)signaling pathway in osteoporotic rats.METHODS:The SD rat model of postmenopausal osteoporosis(PMOP)was established by removing both ovaries.According to the random number table method,the rats were divided into the sham operation group(sham group),PMOP rat model group(PMOP group),Yishen Bugu decoction L group,M group and H group(treated with Yishen Bugu decoction 2,4 and 8 mL/kg by gavage,respectively),and positive control group(treated with estradiol valerate by gavage).The preoperative and postoperative body mass of rats in each group were compared,and the general state of rats in each group was observed.The content of total alkaline phosphatase(TALP)and serum anti-tartrate acid phosphatase(TRAP)in serum were detected by enzyme-linked immunosorbent assay.Rat bone mineral density was detected by dual-energy X-ray.Hematoxylin and Eeosin staining was used to detect the histomorphological changes of rat femur,and spectrophotometer and radioimmunoassay were used to detect the content of NO and cGMP in rat femoral tissue.The protein expressions of Wnt3a andβ-catenin in femur tissue were detected by Western blotting.RESULTS:Compared with the sham group,the body mass of the PMOP group decreased significantly(P<0.05);compared with the PMOP group,the body mass of the Yishen Bugu decoction L,M,and H groups increased significantly(P<0.05),with statistically significant differences.The serum TALP content and femoral BMD of the PMOP group were lower than those of the sham group,and the TRAP content was higher than that of the sham group(P<0.05);the TALP content and femoral BMD in the Yishen Bugu decoction L,M,and H groups were higher than those in the PMOP group,the TRAP content was lower than the PMOP group(P<0.05),with concentration-dependent manner,the differences were statistically significant.There was no significant difference in serum TALP,TRAP content and femoral BMD between Yishen Bugu decoction H group and positive control group(P>0.05).Compared with the sham group,the trabecular bone space of rats in the PMOP group became larger and irregular,the thickness was uneven,and the trabecular bone phenotype was thinner.Compared with the PMOP group,the trabecular bone gap of the rats in the Yishen Bugu decoction L,M,and H groups and positive control group decreased to varying degrees,and the thickness of the trabecular bone was uniform and gradually regular.Compared with the sham group,the contents of NO and cGMP in the femoral tissue of the PMOP group increased significantly(P<0.05);the contents of NO and cGMP in the femoral tissue of Yishen Bugu decoction L,M and H group were lower than those of PMOP group(P<0.05),the differences were statistically significant.Compared with the sham group,the expressions of Wnt3a andβ-catenin proteins in the femoral tissue of the PMOP group were decreased significantly(P<0.05);the expressions of Wnt3a andβ-catenin proteins in the femoral tissue of the Yishen Bugu decoction L,M and H group were higher than those of the PMOP group(P<0.05),with dose-dependent manner,the differences were statistically significant.CONCLUSIONS:Yishen Bugu decoction can promote bone formation in PMOP rats,possibly by inhibiting the NO-cGMP signaling pathway to inhibit bone resorption.
作者
王勤俭
张睿昕
WANG Qinjian;ZHANG Ruixin(Dept.of Osteopathy,Henan Provincial Hospital of Traditional Chinese Medicine/the Second Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)
出处
《中国医院用药评价与分析》
2023年第1期21-26,共6页
Evaluation and Analysis of Drug-use in Hospitals of China
基金
河南省中医药科学研究专项课题(No.20-21ZY2120,No.2019JDZX055)。
关键词
益肾补骨汤
骨质疏松症
骨形成
一氧化氮-环磷酸鸟苷信号通路
Yishen Bugu decoction
Osteoporosis
Bone formation
Nitric oxide-cyclic guanosine monophosphate signaling pathway
