Third Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210028, PR China; Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, PR China.
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, PR China.
J Ethnopharmacol. 2021 Jun 12;273:113996. doi: 10.1016/j.jep.2021.113996. Epub 2021 Mar 6.
Bu Shen Huo Xue Tang (BSHXT) is a traditional Chinese medicine formula that is clinically used in the treatment of premature ovarian insufficiency (POI). However, its therapeutic mechanism remains unclear.
This study aimed to investigate the underlying molecular mechanism of pharmacological activity of BSHXT, via the Nrf2/Keap1 signaling pathway, in the treatment of autoimmune POI.
The chemical composition of BSHXT was analyzed using ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry. The autoimmune POI mouse model was induced by immunizing mice twice, with zona pellucida (ZP) glycoprotein 3 antigen. The autoimmune POI mice were continuously administered BSHXT for 28 days. Body weight and organ indices were recorded. The pathological morphology of the ovaries was observed. The estrous cycle of each mouse was recorded. Immunofluorescence assay was used to detect the levels of ZP antibodies in the mouse ovaries. The levels of ZP antibodies, follicle-stimulating hormone (FSH), anti-Müllerian hormone (AMH), estradiol (E), luteinizing hormone (LH), superoxide dismutase (SOD), and malondialdehyde (MDA) were measured. The expression of genes and proteins involved in the Nrf2/Keap1 signaling pathway were measured by Q-PCR and IHC, respectively.
Twenty-one compounds were identified in the BSHXT water extract. BSHXT was found to increase the body weight and ovarian index, improve ovarian function, and reduce disorders in the estrus cycle. It also reduced the expression of ZP antibodies in the ovaries and serum of POI mice. BSHXT significantly increased E and AMH levels and decreased FSH and LH levels. It also increased the levels of SOD, and reduced MDA levels. The levels of Nrf2 and Keap1 were also increased, and the expression of Nrf2, HO-1 and NQO1 genes was upregulated.
BSHXT has a therapeutic effect on autoimmune POI in mice, which may be a result of the enhanced antioxidant capacity and activation of the Nrf2/Keap1 signaling pathway. BSHXT is a good drug candidate for use as a protective agent for POI and may be used in clinical practice.
补肾活血清汤（BSHXT）是一种临床用于治疗卵巢早衰（POI）的中药方剂。然而，其治疗机制尚不清楚。
本研究旨在通过 Nrf2/Keap1 信号通路探讨 BSHXT 治疗自身免疫性 POI 的药理作用的潜在分子机制。
采用超高效液相色谱-四级杆飞行时间串联质谱联用技术分析 BSHXT 的化学成分。采用两次免疫小鼠透明带 3 糖蛋白抗原诱导自身免疫性 POI 小鼠模型。连续给予 BSHXT 治疗 28 天。记录体重和器官指数。观察卵巢的病理形态。记录每只小鼠的动情周期。免疫荧光法检测小鼠卵巢中透明质酸抗体的水平。检测透明质酸抗体、卵泡刺激素（FSH）、抗苗勒管激素（AMH）、雌二醇（E）、促黄体生成素（LH）、超氧化物歧化酶（SOD）和丙二醛（MDA）水平。通过 Q-PCR 和免疫组化分别检测 Nrf2/Keap1 信号通路相关基因和蛋白的表达。
BSHXT 水提取物中鉴定出 21 种化合物。BSHXT 可增加体重和卵巢指数，改善卵巢功能，减少动情周期紊乱。它还降低了 POI 小鼠卵巢和血清中透明质酸抗体的表达。BSHXT 显著提高了 E 和 AMH 水平，降低了 FSH 和 LH 水平。它还增加了 SOD 水平，降低了 MDA 水平。Nrf2 和 Keap1 的水平也增加了，Nrf2、HO-1 和 NQO1 基因的表达上调。
BSHXT 对小鼠自身免疫性 POI 具有治疗作用，这可能是由于抗氧化能力增强和 Nrf2/Keap1 信号通路激活所致。BSHXT 是治疗 POI 的一种良好候选药物，可用于临床实践。
