| 杨佳迪,石丹宁,陈 瑶,何悦双,赵丕文.四物汤在卵巢衰老大鼠骨骼肌中的雌激素样作用机制研究[J].,2023,(4):607-612 |
| 四物汤在卵巢衰老大鼠骨骼肌中的雌激素样作用机制研究 |
| Study on the Estrogen-like Action Mechanism of Siwu Decoction in Skeletal Muscle of Ovarian Aging Rats |
| 投稿时间：2022-07-22 修订日期：2022-08-18 |
| DOI：10.13241/j.cnki.pmb.2023.04.002 |
| 中文关键词: 四物汤 肌肉减少症 IGF-1/PI3K/AKT/mTOR 4-乙烯基环己烯二环氧化合物 植物雌激素 |
| 英文关键词: Siwu Decoction Sarcopenia IGF-1/PI3K/AKT/mTOR VCD Phytoestrogens |
| 基金项目:国家自然科学基金项目（81673764） |
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| 中文摘要: |
| 摘要 目的：探讨四物汤通过胰岛素样生长因子-1（Insulin-ike growth factors-1，IGF-1）/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素靶蛋白（Mammalian target of rapamycin，mTOR）mTOR信号通路发挥对4-乙烯基环己烯二环氧化合物（4-Vinylcyclohexene Diepoxide，VCD）诱导的卵巢衰老大鼠骨骼肌的保护作用及其分子机制。方法：选用28日龄雌性F-344大鼠，随机选取6只作为空白对照组，剩余大鼠连续腹腔注射VCD溶液（160 mg/kg/d）20天，每天阴道涂片检测动情周期，连续观察12 d无角化细胞或仅有少量角化细胞即符合"卵巢衰老"表现，经动情周期筛选出24只造模成功的大鼠，分为模型组（Model）、阳性（戊酸雌二醇，E2）对照组、四物汤高剂量（SWT-H）组和四物汤低剂量（SWT-L）组，每组6只，灌胃持续3周后取材。取大鼠骨骼肌组织进行HE染色观察病理组织结构；MASSON染色骨骼肌纤维形态变化；RT-PCR检测骨骼肌组织中各基因的mRNA水平。结果：与空白组比较，模型组骨骼肌肌纤维排列疏松，分布不均且有断点，胞浆不均，细胞核增多，出现增生的结缔组织，胶原纤维逐渐增多且肌纤维间距变宽；与模型组相比，E2组、SWT-H组与SWT-L组肌纤维排列相对整齐，胞浆较为均匀。肌肉形态较规则，纤维间距缩短，胶原纤维减少。RT-PCR结果显示，与空白组相比，模型组IGF-1、PI3K、AKT、mTOR基因的mRNA表达量明显下调，E2组、SWT-H组与SWT-L组的IGF-1、PI3K、mTOR的表达明显上升，SWT-H组AKT表达无明显变化，无统计学意义，SWT-L组AKT表达相对降低。与模型组相比，E2组、SWT-H组与SWT-L组的IGF-1、AKT、mTOR的表达明显增加，且具有剂量依赖性，PI3K表达增加，但无明显剂量依赖性。结论：四物汤可以明显改善VCD诱导的卵巢衰老大鼠的肌肉减少情况，其机制可能是通过IGF-1-PI3K-AKT-mTOR信号通路来发挥其抑制肌肉流失的作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of Siwu Decoction through insulin-like growth factor-1 (Insulin-like growth factors-1, IGF-1)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/rapamycin target protein (Mammalian target of rapamycin, mTOR) mTOR signaling pathway exerts the protective effect of 4-vinylcyclohexene diepoxide (4-Vinylcyclohexene Diepoxide, VCD)-induced ovarian aging rat skeletal muscle and its molecular mechanism. Methods: 28-Day-old female F-344 rats were used, and randomly selected six rats as control group. The remaining rats were intraperitoneally injected with VCD solution (160 mg/kg/d) for 20 days, the estrous cycle was detected by vaginal smear every day, and no keratinocytes or only a small amount of keratinization were observed continuously for 12 days. The cells are in line with the performance of "ovarian aging". 24 rats were successfully modeled after estrous cycle screening, and divided to model group (Model), positive (estradiol valerate, E2) control group, Siwu Decoction high dose (SWT-H) group, and Siwu Decoction low dose (SWT-L) group with six rats in each group. The gavage was taken for three weeks. The skeletal muscle tissue was acquired for HE staining to observe the pathological structure; MASSON staining was used for morphological changes of skeletal muscle fibers; RT-PCR was used to detect the mRNA levels of each gene in skeletal muscle tissue. Results: Compared with the blank group, the skeletal muscle fibers in the model group were loosely arranged, unevenly distributed with breakpoints and uneven cytoplasm, increased nuclei, appeared hyperplastic connective tissue, and gradually increased collagen fibers, and the muscle fiber spacing was wider. Compared with the model group, the muscle fibers of E2 group, SWT-H group, and the SWT-L group were arranged neatly, and the cytoplasm was relatively uniformed. The shape of muscles was more regular, fiber spacing was shortened, and collagen fibers were reduced. RT-PCR results showed that the mRNA expressions of IGF-1, PI3K, AKT, and mTOR were significantly reduced in the model group compared with the blank group. The expressions of IGF-1, PI3K, and mTOR in the E2 group, SWT-H group and SWT-L group were significantly increased. The expression of AKT in the SWT-H group had no significant change, and there was no statistical significance. The expression of AKT in the SWT-L group was relatively decreased. Compared with the model group, the expressions of IGF-1, AKT and mTOR in the E2 group, the SWT-H group, and the SWT-L group were significantly increased, and the SWT group had a dose-dependent increase. The expression of PI3K was increased, but there was no significant dose-dependent. Conclusion: Siwu Decoction can significantly improve the sarcopenia in VCD-induced ovarian aging rats, and the mechanism may be through the IGF-1-PI3K-Akt-mTOR signaling pathway to exert its effect on inhibiting muscle loss. |
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