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王桂云,吴结枝,雷晓明,李荣慧,曹宝丹,刘慧萍,张国民.壮骨止痛胶囊对去卵巢大鼠MAPK信号通路的影响[J].湖南中医药大学学报英文版,2021,41(2):205-210.[Click to copy
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| 壮骨止痛胶囊对去卵巢大鼠MAPK信号通路的影响 |
| 王桂云,吴结枝,雷晓明,李荣慧,曹宝丹,刘慧萍,张国民 |
| (湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察壮骨止痛胶囊对去卵巢大鼠骨组织MAPK信号通路的影响，探讨其治疗绝经后骨质疏松的机制。方法 将36只SPF级SD雌性大鼠随机分为6组，每组6只。随机选定2组为空白组和假手术组，其余大鼠均进行切除卵巢造模处理，即采用国内外公认的去除雌性大鼠双侧卵巢方法建立绝经后骨质疏松病理模型，将造模成功的24只大鼠依次分为模型组、壮骨止痛胶囊组、雷帕酶素（rapamycin，RAPA）组和3-甲基腺嘌呤（3-methyladenine，3-MA）组。术后第6天开始灌胃给药，连续3个月。末次灌胃给药后第2天开始取材。HE染色观察骨组织病理形态；ELISA检测血清雌二醇（estradiol，E2）的含量；免疫组化法分析股骨ERK1/2、p-ERK1/2、p38 MAPK、p-p38 MAPK蛋白的表达。结果 （1）空白组和假手术组骨髓腔无明显变化，模型组大鼠骨髓腔明显扩大，壮骨止痛胶囊组和RAPA组大鼠骨髓腔扩大不明显。（2）空白组和假手术组的血清E2含量差异无统计学意义（P>0.05）；与假手术组相比，模型组血清E2值降低，且有显著性差异（P<0.01）；与模型组相比，壮骨止痛胶囊组和RAPA组血清E2值升高、3-MA组血清E2值降低，差异均具有统计学意义（P<0.01）；与3-MA组相比，壮骨止痛胶囊组和RAPA组大鼠E2值升高，差异具有统计学意义（P<0.01）。（3）空白组和假手术组骨组织ERK1/2、P38MAPK及其磷酸化蛋白表达差异无统计学意义（P>0.05）；与假手术组相比，模型组骨组织的ERK1/2、P38 MAPK及其磷酸化蛋白表达降低，差异具有统计学意义（P<0.01）；与模型组相比，壮骨止痛胶囊组和RAPA组ERK1/2、P38 MAPK及其磷酸化蛋白表达升高（P<0.01），3-MA组ERK1/2、P38 MAPK蛋白表达降低（P<0.01），差异均具有统计学意义；与3-MA组相比，壮骨止痛胶囊组和RAPA组的ERK1/2、P38 MAPK及其磷酸化蛋白表达升高，差异具有统计学意义（P<0.01），且壮骨止痛胶囊组和RAPA组股骨组织ERK1/2、P38 MAPK及其磷酸化蛋白表达差异无统计学意义（P>0.05）。结论 壮骨止痛胶囊能升高血清中E2的表达，可能通过上调去势大鼠骨组织中ERK1/2、p-ERK1/2、p38 MAPK及p-p38 MAPK蛋白表达来调控MAPK信号通路。 |
| 关键词: 绝经后骨质疏松症 壮骨止痛胶囊 MAPK信号通路 雌二醇 |
| DOI：10.3969/j.issn.1674-070X.2021.02.009 |
| Received:September 04, 2020 |
| 基金项目:国家自然科学基金面上项目（81973920）；湖南省自然科学基金面上项目（2018JJ2297）；湖南省教育厅科学研究重点项目（2019A370）；湖南省大学生研究性学习和创新性实验计划项目（2017281，2018420）。 |
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| Effect of Zhuanggu Zhitong Capsule on MAPK Signal Pathway in Ovariectomized Rats |
| WANG Guiyun,WU Jiezhi,LEI Xiaoming,LI Ronghui,CAO Baodan,LIU Huiping,ZHANG Guomin |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effect of Zhuanggu Zhitong Capsule on MAPK signaling pathway in bone tissue of ovariectomized rats, and to explore the mechanism in treating postmenopausal osteoporosis. Methods 36 SPF female SD rats were randomly divided into 6 groups, with 6 rats in each group. Two groups were randomly selected as the blank group and the sham operation group, the other rats were ovariectomized to establish the pathological model of postmenopausal osteoporosis by removing bilateral ovaries of female rats, 24 rats with successful model were randomly divided into the model group, the Zhuanggu Zhitong Capsule group, the rapamycin (RAPA) group and the 3-methyladenine (3-MA) group. The drug was administered 6 days after operation for 3 months. The samples were taken from the second day after the last intragastric administration. The pathological morphology of bone tissue was observed by HE staining; the content of serum estradiol (E2) was detected by ELISA; the expressions of ERK1/2 p-ERK1/2, p38 MAPK and P-P38 MAPK were analyed by immunohistochemistry. Results (1) There was no significant change in the bone marrow cavity of the blank group and the sham operation group. The bone marrow cavity of the model group was significantly enlarged, while that of the Zhuanggu Zhitong Capsule group and the RAPA group was not. (2) Compared with the blank group, the level of serum E2 in the sham operation group was not statistically significant (P>0.05). Compared with the sham operation group, the level of serum E2 in the model group was significantly lower, and there was a significant difference (P<0.01). Compared with the model group, the serum E2 value of Zhuanggu Zhitong Capsule group and the RAPA group increased, and the serum E2 value of the 3-MA group decreased, with significant differences (P<0.01). Compared with the 3-MA group, the E2 value in the Zhuanggu Zhitong Capsule group and the RAPA group increased, there were significant differences (P<0.01). (3) There were no significant difference in the expressions of ERK1/2, p38 MAPK and phosphorylated protein of bone tissue between the sham operation group and the blank group (P>0.05). Compared with the sham operation group, the expressions of ERK1/2, p38 MAPK and their phosphorylated protein of bone tissue in the model group were significantly lower, and the differences were statistically significant (P<0.01). Compared with the model group, the expressions of ERK1/2, p38 MAPK and their phosphorylated proteins in the Zhuanggu Zhitong Capsule group and the RAPA group were significantly higher, and the difference were statistically significant (P<0.01), and the expressions of ERK1/2, p38 MAPK and their phosphorylated proteins in 3-MA group were significantly lower, and the difference were statistically significant (P<0.01). Compared with the 3-MA group, the expressions of ERK1/2, p38 MAPK and their phosphorylated proteins in the Zhuanggu Zhitong Capsule group and the RAPA group were significantly higher, and the difference were statistically significant (P<0.01), there were no significant difference in the xpressions of ERK1/2, p38 MAPK and phosphorylated protein between the Zhuanggu Zhitong Capsule group and the RAPA group (P>0.05). Conclusion Zhuanggu Zhitong Capsule can increase the expression of E2 in serum, and regulate MAPK signaling pathway by up regulating the protein expressions of ERK1/2, p-ERK1/2, p38 MAPK and P-P38 MAPK in bone tissue of ovariectomized rats. |
| Key words: postmenopausal osteoporosis Zhuanggu Zhitong Capsule MAPK signaling pathway estradiol |
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