Li Shan-Shan, Zhang Hai-Zhu, Zhang Long, Chen Shuai-Shuai, He Xian, Xiao Xiao-He, Wang Jia-Bo, Niu Ming
Department of Pharmacy and Chemistry, Dali University Dali 671000, China China Military Institute of Chinese Medicine, Fifth Medical Center, Chinese PLA General Hospital Beijing 100039, China.
Department of Pharmacy and Chemistry, Dali University Dali 671000, China.
Zhongguo Zhong Yao Za Zhi. 2021 May;46(10):2556-2564. doi: 10.19540/j.cnki.cjcmm.20201014.201.
Based on the heat-clearing and detoxifying effects of Gentianae Radix et Rhizoma, the network pharmacology is mainly used to predict the potential targets of Gentianae Radix et Rhizoma for anti-inflammatory activity and to perform the experimental verification. A method for detecting the biological potency of Gentianae Radix et Rhizoma based on verifiable targets has been established to provide a reference for improving the quality evaluation and control standards of Gentianae Radix et Rhizoma. High performance liquid chromatography can be used to construct chemical fingerprints of different batches of Gentianae Radix et Rhizoma. Constructing a component-target-disease network of Gentianae Radix et Rhizoma for its anti-inflammatory activity was applied to screen potential anti-inflammatory components and related targets of Gentianae Radix et Rhizoma, and to verify the target of Gentianae Radix et Rhizoma by using biological evaluation methods. Detecting the biological potency of different batches of Gentianae Radix et Rhizoma extracts was used to inhibit COX-2 enzyme activity based the verifiable target cyclooxygenase-2(COX-2). The results showed that different batches of Gentianae Radix et Rhizoma accorded with the pharmacopoeia testing regulations, and the chemical fingerprints have a high similarity(similarity>0.93), suggesting that there is no significant difference in the characteristics of the chemical components. Based on network pharmacology predictions, 18 candidate targets were found to have potential direct interactions with the ingredients in Gentianae Radix et Rhizoma. Among them, the most important target is COX-2. Based on the experimental verification of recombinant human COX-2 protease activity inhibition, Gentianae Radix et Rhizoma can inhibit the COX-2 enzyme activity in a dose-dependent manner. It can function with a low concentration(0.75 mg·mL(-1)), which preliminarily confirmed the accuracy of network pharmacology prediction. The biological potency detection method of Gentianae Radix et Rhizoma based on COX-2 inhibitory activity was optimized and established. The qualitative response parallel line method was used to calculate the biological potency of anti-inflammatory activity, which ranged from 23.04 to 46.60 U·mg(-1). For network pharmacology prediction, it can screen and clarify the possible targets of traditional Chinese medicine rapidly, which can guide the establishment of a biological evaluation method for the quality of medicinal materials with related activities. Compared with chemical fingerprints, the biological potency testing can better detect quality fluctuations of traditional Chinese medicine.
基于龙胆的清热燥湿、泻肝胆火功效,运用网络药理学预测龙胆抗炎活性的潜在靶点并进行实验验证。建立了一种基于可验证靶点检测龙胆生物效价的方法,为提高龙胆质量评价与控制标准提供参考。采用高效液相色谱法构建不同批次龙胆的化学指纹图谱。构建龙胆抗炎活性的成分 - 靶点 - 疾病网络,筛选龙胆潜在的抗炎成分及相关靶点,并运用生物学评价方法验证龙胆的靶点。基于可验证靶点环氧化酶 -2(COX -2)检测不同批次龙胆提取物的生物效价,以抑制COX -2酶活性。结果表明,不同批次龙胆符合药典检测规定,化学指纹图谱相似度高(相似度>0.93),表明化学成分特征无显著差异。基于网络药理学预测,发现18个候选靶点与龙胆中的成分可能存在直接相互作用。其中,最重要的靶点是COX -2。通过重组人COX -2蛋白酶活性抑制实验验证,龙胆能以剂量依赖性方式抑制COX -2酶活性。在低浓度(0.75 mg·mL⁻¹)下即可发挥作用,初步证实了网络药理学预测的准确性。优化并建立了基于COX -2抑制活性的龙胆生物效价检测方法。采用定性反应平行线法计算抗炎活性的生物效价,范围为23.04至46.60 U·mg⁻¹。对于网络药理学预测,它能快速筛选并明确中药可能的靶点,可指导建立具有相关活性的药材质量生物学评价方法。与化学指纹图谱相比,生物效价检测能更好地检测中药质量波动情况。