Department of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, PR China; Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui Academy of Chinese Medicine, Hefei 230012, PR China.
Department of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, PR China; Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine, Hefei 230012, PR China; Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui Academy of Chinese Medicine, Hefei 230012, PR China.
J Pharm Biomed Anal. 2022 Aug 5;217:114828. doi: 10.1016/j.jpba.2022.114828. Epub 2022 May 10.
Clinopodium chinense (Benth.) O. Kuntze (C. chinense), a traditional Chinese medicine with significant astringent and hemostatic properties, is mainly used to treat abnormal uterine bleeding (AUB) with remarkable curative effect, but the active ingredients of which remain unclear. This study aimed to screen and identify the main anti-AUB components of C. chinense via spectrum-effect relationship analysis and experiment validation. Firstly, total extract of C. chinense (TEC) of 12 batches samples was prepared by Chinese Pharmacopoeia. The fingerprint chromatogram of TEC was established by UPLC-Q-TOF-MS. The AUB model was established by intragastric administration of mifepristone and misoprostol to pregnant rats, followed by the treatment with TEC. After drug administration lasting 7 days, metrorrhagia volume was measured, pathological changes in uterine tissue were evaluated by HE staining, the levels of TXB-2, TNF-α, and IL-6 were measured by ELISA. The spectrum-effect relationship was investigated by grey relational analysis (GRA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Finally, the potential active ingredients of TEC screened by spectrum-effect relationship analysis were subsequently verified both in vitro and in vivo. A total of 25 common peaks were obtained from the fingerprint chromatogram of the 12 bathes TEC samples, 12 peaks were identified according to the reference substances. Comparing with the model group, TEC significantly reduced the uterine bleeding volume, alleviated endometrial injury, increased plasma TXB2 level, and decreased plasma IL-6 and TNF-α levels. Furthermore, seven components including kaempferol, quercetin, buddlejasaponin Ⅳb, hesperidin, naringenin, apigenin, and saikosaponin a were identified via spectrum-effect relationship analysis. In vitro and in vivo verification indicated that buddlejasaponin Ⅳb, hesperidin, naringenin, apigenin, and saikosaponin a were responsible for the anti-AUB activity of TEC. In conclusion, the present study established a spectrum-effect relationship for C. chinense and identified the main anti-AUB compounds in TEC, which provides insight for the exploration of bioactive components and quality control of C. chinense.
荆草（Benth.）O. Kuntze（C. chinense），一种具有显著收敛和止血特性的传统中药，主要用于治疗异常子宫出血（AUB），疗效显著，但其中的活性成分仍不清楚。本研究旨在通过谱效关系分析和实验验证筛选和鉴定荆草的主要抗 AUB 成分。首先，按照《中国药典》制备 12 批荆草总提取物（TEC）。采用 UPLC-Q-TOF-MS 建立 TEC 的指纹图谱。通过给怀孕大鼠灌胃米非司酮和米索前列醇建立 AUB 模型，然后用 TEC 治疗。连续给药 7 天后，测量出血量，HE 染色评估子宫组织病理变化，ELISA 法测定 TXB-2、TNF-α和 IL-6 水平。采用灰色关联度分析（GRA）和正交偏最小二乘判别分析（OPLS-DA）研究谱效关系。最后，通过谱效关系分析筛选出 TEC 的潜在活性成分，随后进行体内外验证。从 12 批 TEC 样品的指纹图谱中得到 25 个共有峰，根据对照品鉴定出 12 个峰。与模型组相比，TEC 能显著减少子宫出血量，减轻子宫内膜损伤，增加血浆 TXB2 水平，降低血浆 IL-6 和 TNF-α水平。此外，通过谱效关系分析鉴定出 7 种成分，包括山奈酚、槲皮素、梓醇苷 IVb、橙皮苷、柚皮苷、芹菜素和柴胡皂苷 a。体内外验证表明，梓醇苷 IVb、橙皮苷、柚皮苷、芹菜素和柴胡皂苷 a 是 TEC 抗 AUB 活性的主要成分。总之，本研究建立了荆草的谱效关系，鉴定了 TEC 中主要的抗 AUB 化合物，为荆草生物活性成分的探索和质量控制提供了依据。
