基于Notch1-Hes1-Prdx蛋白家族通路探究强骨胶囊治疗绝经后骨质疏松的炎症及氧化应激机制

doi:10.20047/j.issn1673-7210.2023.23.02

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摘要目的基于Notch1-Hes1-Prdx蛋白家族通路探究强骨胶囊治疗绝经后骨质疏松（PMOP）的炎症及氧化应激机制。方法 60只5月龄清洁级雌性SD大鼠，体重（300±20）g，采用随机数字表法分为假手术组，模型对照组，阳性对照组及强骨胶囊低、中、高剂量组，每组10只。除假手术组外，其余大鼠通过切除双侧卵巢建立PMOP模型，假手术组仅切除双侧卵巢周边脂肪组织。阳性对照组经皮下注射200 μg/kg雌二醇，2次/周；强骨胶囊低、中、高剂量组分别予以100、200、300 mg/（kg·d）强骨胶囊灌胃；假手术组及模型对照组灌胃等量生理盐水，连续治疗12周。比较各组大鼠股骨骨密度（BMD）值，血浆炎症标志物[白细胞介素（IL）-1β、IL-6、IL-8、IL-17、肿瘤坏死因子-α（TNF-α）]、氧化应激标志物水平[单胺氧化酶A（MAOA）、总抗氧化能力（T-AOC）、高级氧化蛋白产物（AOPP）、超氧化物歧化酶（SOD）]及Jagged1、Notch1、Hes1、Prdx蛋白家族（Prdx1~6）的蛋白表达水平；RT-qPCR及Western blot测定外周血单个核细胞（PBMC）及骨组织中的Jagged1、Notch1、Hes1及Prdx1~6 mRNA及蛋白表达。结果与假手术组比较，模型对照组大鼠BMD值、T-AOC、SOD水平降低，IL-1β、IL-6、IL-8、IL-17、TNF-α、MAOA、AOPP水平升高；血浆、PBMC及骨组织中Jagged1、Notch1、Hes1表达水平升高，Prdx1、Prdx6表达水平降低（P＜0.05）。与模型对照组比较，强骨胶囊中、高剂量组大鼠BMD值、T-AOC和SOD水平升高，IL-1β、IL-6、IL-8、IL-17、TNF-α、MAOA、AOPP水平降低；血浆、PBMC及骨组织中Jagged1、Notch1、Hes1表达水平降低，Prdx1、Prdx6表达水平升高（P＜0.05）。与强骨胶囊低剂量组比较，强骨胶囊中、高剂量组大鼠股骨BMD值、T-AOC和SOD水平升高（P＜0.05），IL-1β、IL-6、IL-8、IL-17、TNF-α、MAOA、AOPP水平降低；血浆、PBMC及骨组织中Jagged1、Notch1、Hes1表达水平降低，Prdx1、Prdx6表达水平升高（P＜0.05）。与强骨胶囊中剂量组比较，强骨胶囊高剂量组大鼠股骨BMD值、T-AOC和SOD水平升高，IL-1β、IL-6、IL-8、IL-17、TNF-α、MAOA、AOPP水平降低；血浆、PBMC及骨组织中Jagged1、Notch1、Hes1表达水平降低，Prdx1、Prdx6表达水平升高（P＜0.05）。各组血浆、PBMC及骨组织中Prdx2~5表达水平比较，差异无统计学意义（P＞0.05）。结论强骨胶囊可能是通过抑制Notch1-Hes1-Prdx信号通路下调Jagged1、Notch1、Hes1表达，上调Prdx1、Prdx6表达，增强机体抗氧化应激能力并改善炎症反应发挥抗骨质疏松作用，其中强骨胶囊作用呈剂量依赖性。

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	鲁林1　　方虹2

关键词 ：强骨胶囊, Notch1-Hes1-Prdx蛋白家族通路, 绝经后骨质疏松, 炎症, 氧化应激

Abstract：

Objective To explore the mechanism of inflam- mation and oxidative stress of Qianggu Capsules in the treatment of postmenopausal osteoporosis (PMOP) based on Notch1- Hes1-Prdx protein family pathway. Methods Sixty five- month-old specific pathogen free female SD rats with body mass of (300 ± 20) g were divided into sham operation group, model control group, positive control group, and Qianggu Capsules low, medium, and high dose groups of by random number table method, with 10 rats in each group. Except for the sham operation group, the PMOP model was established by bilateral ovaries resection, and only peripheral adipose tissue of bilateral ovaries was removed in the sham operation group. The positive control group was subcutaneously injected with 200 μg/kg Estradiol, twice a week. Qianggu Capsules low, medium, and high dose groups were given 100, 200, and 300 mg/(kg·d) Qianggu Capsules by gavage, respectively; the sham operation group and model control group was given the same amount of physiological saline by gavage, the treatment lasted for 12 weeks. Bone mineral density (BMD) value, plasma inflammatory markers (interleukin [IL]-1β, IL-6, IL-8, IL-17, tumor necrosis factor-α [TNF-α]), oxidative stress markers(monoamine oxidase-A [MAOA], total antioxidant capacity [T-AOC], advanced oxidation protein products [AOPP], superoxide dismutase [SOD] ), and the protein expression level of Jagged1, Notch1, Hes1, and Prdx protein family (Prdx1-6) were compared among all groups; the mRNA and protein expressions of Jagged1, Notch1, Hes1, and Prdx1-6 in peripheral blood mononuclear cells (PBMC) and bone tissue were determined by RT-qPCR and Western blot. Results Compared with sham operation group, BMD value, T-AOC, and SOD levels of rats were decreased, IL-1β, IL-6, IL-8, IL-17, TNF-α, MAOA, and AOPP levels of rats were increased; the expression levels of Jagged1, Notch1, and Hes1 in plasma, PBMC, and bone tissue of rats were increased, while the expression levels of Prdx1 and Prdx6 of rats were decreased in model control group (P＜0.05). Compared with model control group, BMD value, T-AOC, and SOD levels of rats were increased, IL-1β, IL-6, IL-8, IL-17, TNF-α, MAOA, and AOPP levels of rats were decreased; the expression levels of Jagged1, Notch1, and Hes1 in plasma, PBMC, and bone tissue of rats were decreased, while the expression levels of Prdx1 and Prdx6 of rats were increased in Qianggu Capsules medium and high dose groups (P＜0.05). Compared with Qiangu Capsules low dose group, BMD value, T-AOC, and SOD levels of rats were increased, while IL-1β, IL-6, IL-8, IL-17, TNF-α, MAOA, and AOPP levels of rats were decreased; the expression levels of Jagged1, Notch1, and Hes1 in plasma, PBMC, and bone tissue of rats were decreased, while the expression levels of Prdx1 and Prdx6 of rats were increased in Qianggu Capsules medium and high dose groups (P＜0.05). Compared with Qianggu Capsules medium dose group, BMD value, T-AOC, and SOD levels of rats were increased, IL-1β, IL-6, IL-8, IL-17, TNF-α, MAOA, and AOPP levels of rats were decreased; the expression levels of Jagged1, Notch1, and Hes1 in plasma, PBMC, and bone tissue of rats were decreased, while the expression levels of Prdx1 and Prdx6 of rats were increased in Qiangu Capsules highdose group (P＜0.05). There were no significant differences in PRDX2-5 expressions in plasma, PBMC, and bone tissue of rats among all groups (P＞0.05). Conclusion Qianggu Capsules may down-regulate the expressions of Jagged1, Notch1, and Hes1 and up-regulate the expressions of Prdx1 and Prdx6 by inhibiting the Notch1-Hes1-Prdx signaling pathway, thus enhancing the antioxidant stress ability and improving the inflammatory response to play an anti-osteoporosis role , and the effect of Qianggu Capsules were dose-dependent.

Key words： Qianggu Capsules Notch1-Hes1-Prdx protein family pathway Postmenopausal osteoporosis Inflammation Oxidative stress

基金资助:湖北省自然科学基金创新发展联合基金项目（2022CFD150）；
湖北省中医药管理局中医药科研项目（ZY2023F009）；
湖北省武汉市卫生健康委员会科研计划资助项目（WZ20Q09）。

通讯作者: 方虹（1983.2-），女，硕士，副主任医师，主要从事中西结合临床研究。

作者简介: 鲁林（1981.12-），男，硕士，副主任医师，主要从事中西结合骨科研究。

引用本文:

鲁林1　　方虹2. 基于Notch1-Hes1-Prdx蛋白家族通路探究强骨胶囊治疗绝经后骨质疏松的炎症及氧化应激机制[J]. 中国医药导报, 2023, 20(23): 8-15.
LU Lin1 FANG Hong2. Exploration on the mechanism of inflammation and oxidative stress of Qianggu Capsules in the treatment of postmenopausal osteoporosis based on the Notch1-Hes1-Prdx protein family pathway. 中国医药导报, 2023, 20(23): 8-15.

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