骨碎补总黄酮对骨质疏松大鼠骨组织中硬化蛋白表达的影响及其作用机制

doi:10.13481/j.1671-587x.20200503

摘要/Abstract

摘要： 目的：探讨骨碎补总黄酮对骨质疏松大鼠骨组织中骨硬化蛋白（SOST）表达的影响，阐明骨碎补总黄酮抗骨质疏松的作用机制。方法：将40只8月龄雌性大鼠分为正常对照组（每天灌胃生理盐水）、骨质疏松模型组（90 mg·kg^-1·d^-1维甲酸灌胃给药，持续14 d）、阳性药组（维甲酸灌胃14 d，同时以0.1 mg·kg^-1·d^-1皮下注射阿仑膦酸钠30 d）和骨碎补总黄酮组（维甲酸灌胃14 d，同时以58 mg·kg^-1·d^-1骨碎补总黄酮灌胃给药30 d），每组10只。采集各组大鼠静脉血，检测血清中钙、磷和碱性磷酸酶（ALP）水平，肝和肾功能指标，包括丙氨酸氨基转移酶（ALT）、门冬氨酸氨基转移酶（AST）、肌酐（Crea）和尿素氮（BUN）；取大鼠一侧股骨，采用HE染色观察骨组织病理形态表现；采用qRT-PCR法检测各组大鼠骨组织中SOST mRNA表达水平，Western blotting法检测大鼠骨组织中SOST蛋白表达水平。结果：与骨质疏松模型组比较，阳性药组大鼠血清中钙和磷水平明显降低（P<0.05或P<0.01），而骨碎补总黄酮组大鼠血清中钙水平明显升高（P<0.05）；与阳性药组比较，骨碎补总黄酮组大鼠血清中钙、磷和ALP水平明显升高（P<0.05或P<0.01）。与正常对照组比较，骨质疏松模型组、阳性药组和骨碎补总黄酮组大鼠血清中Crea水平明显降低（P<0.01）；与骨质疏松模型组和阳性药组比较，骨碎补总黄酮组大鼠血清中Crea水平明显降低（P<0.01）；各组大鼠血清中ALT、AST和BUN水平比较差异均无统计学意义（P>0.05）。光镜下观察，骨质疏松模型组大鼠骨组织骨小梁稀疏、变薄和断裂；与骨质疏松模型组比较，阳性药组和骨碎补总黄酮组大鼠骨小梁明显变厚、断裂减少。与骨质疏松模型组比较，阳性药组和骨碎补总黄酮组大鼠骨组织中SOST mRNA表达水平均明显降低（P<0.01）；阳性药组大鼠骨组织中SOST蛋白表达水平较正常对照组和骨质疏松模型组均明显降低（P<0.01），骨碎补总黄酮组大鼠SOST蛋白表达水平虽然较正常对照组和骨质疏松模型组低，但差异无统计学意义（P>0.05）。结论：骨碎补总黄酮拮抗骨质疏松症的作用机制可能是通过抑制骨细胞合成SOST进而促进骨形成。

关键词: 骨质疏松症, 骨代谢, 骨碎补, 骨硬化蛋白

Abstract: Objective: To analyze the effect of total flavonoids of Rhizomadrynariae on the expression of sclerostin (SOST) in bone tissue of the osteoporosis rats, and to explore the mechanism of tatal flavonoids of Rhzomadrynariae in the anti-osteoporosis process. Methods: A total of 40 female rats aged 8 months were divided into normal control group (fed daily with saline by gavage), osteoporosis model group(administrated with 90 mg·kg^-1·d^-1 retinoic acid by gavaged for 14 d),positive drug group (administrated with retinoic acid by gavage for 14 d and subcutaneously injected with 0.1 mg·kg^-1·d^-1 alendronare for 30 d),and total flavonoid of Rhizomadrynariae group (administrated with retinoic acid by gavaged for 14 d and 58 mg·kg^-1·d^-1 total flavonoids of Rhizomadrynariae by gavage for 30 d);there were 10 rats in each group. The vein blood samples of the rats were collected after administratration, and the levels of calcium, phosphorus and alkaline phosphatase (ALP) and the indexes of liver and kidney function were detected, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (Crea) and blood urea nitrogen (BUN); one sides of the femurs of the rats were taken and the pathomorphology of bone tissue was observed by HE staining. The expression levels of SOST mRNA in bone tissue of the rats in various groups were detected by qRT-PCR and the expression levels of SOST protein in bone tissue of the rats were detected by Western blotting method. Results: Compared with osteoporosis model group, the serum calcium and phosphorus levels of the rats in positive drug group were significantly decreased(P<0.05 or P<0.01), however, the serum calcium level of the rats in total flavonoids of Rhizomadrynariae group was significantly increased (P<0.05); compared with positive drug group, the serum levels of calcium, phosphorus and ALP in total flavonoids of Rhizomadrynariae group were significantly increased (P<0.05 or P<0.01). Compared with normal control group, the serum Crea levels of the rats in osteoporosis model group, positive drug group and total flavonoids of Rhizomadrynariae group were significantly decreased (P<0.01). Compared with osteoporosis model group and positive drug group, the serum Crea level of the rats in total flavonoids of Rhizomadrynariae group was significantly reduced (P<0.01); there were no significant differences in the serum ALT, AST,and BUN levels of the rats among various groups (P>0.05). Under the light microscope, the metaphysis trabeculae of the rats in osteoporosis model group were atrophy, thin and broken, but the trabeculae in positive group and total flavonoids of Rhizomadrynariae group were obviously thickened and continuity enhancement. Compared with osteoporosis model group, the expression levels of SOST mRNA in bone tissue of the rats in positive drug group and total flavonoids of Rhizomadrynariae group were significantly decreased(P<0.01).The expression level of SOST protein in bone tissue of the rats in positive drug group was significantly lower than those in normal control group and osteoporosis model group (P<0.01). The expression level of SOST protein in bone tissue of the rats in total flavonoids of Rhizomadrynariae group was lower than those in normal control group and osteoporosis model group, but the differences were not statistically significant (P>0.05). Conclusion: The mechanism of anti-osteoporosis of total flavonoids of Rhizomadrynariae may be related to inhibiting the synthesis of SOST by osteocytes to promote the bone formation.

Key words: osteoporosis, bone metabolism, Rhizomadrynariae, sclerostin

中图分类号:

R285.5

孙景春, 金辉, 杨雯棋, 徐辉. 骨碎补总黄酮对骨质疏松大鼠骨组织中硬化蛋白表达的影响及其作用机制[J]. 吉林大学学报(医学版), 2020, 46(05): 911-916.

SUN Jingchun, JIN Hui, YANG Wenqi, XU Hui. Effect of total flavonids of Rhizomadrynariae on sclerostin expression in bone tissue of osteoporosis rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 911-916.

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