School of Pharmacy, Guangxi University of Chinese Medicine, Guangxi, 530200, China.
The First Affiliated Hospital of Guangxi Medical University, Guangxi, 530021, China.
Curr Comput Aided Drug Des. 2024;20(1):49-59. doi: 10.2174/1573409919666230417085106.
Caulis Spatholobi is one of the necessary Chinese herbal medicines for hematologists in the treatment of malignant tumors, but its potential targets and molecular mechanisms need further exploration.
This study aimed to predict the relevant targets of the treatment of chronic myeloid leukemia (CML) with Caulis Spatholobi by applying the network pharmacology method, and in vitro cell experiments were conducted to verify the mechanism of Caulis Spatholobi in the treatment of CML.
TCMSP, ETCM, Genecards, and GisGeNET databases were used to obtain relevant targets of Caulis Spatholobi in the treatment of CML. Go and KEGG analyses were performed using the David database. Using Cytoscape 3.7.2, the "active compounds-targets-pathways" network was constructed. Further validation was carried out by pharmacological experiments in vitro. The proliferation and apoptosis of K562 cells were observed by the MTT method and Hoechst 33242 fluorescence staining method. The predicted targets and their related signal pathways were verified by western blotting.
In this study, 18 active compounds and 43 potential targets were obtained. The results of the MTT method showed that compared with the normal control group, 62.5-500 μg/mL alcohol extract of Caulis Spatholobi had an obvious inhibitory effect on K562 and the IC value was less than 100 μg/mL. The Hoechst 33242 fluorescence staining method showed that the alcohol extract of Caulis Spatholobi could promote apoptosis. The results of western blotting showed that compared with the normal control group, the expressions of Bax and Caspase-3 proteins in the 62.5 and 125 μg/mL alcohol extract of Caulis Spatholobi groups were significantly up-regulated (p < 0.05). The expression of Bcl-2 in the 125 μg/mL alcohol extract of the Caulis Spatholobi group was significantly down-regulated (p < 0.01), and the expression of Bcl-2 in the 62.5 and 31.25 μg/mL alcohol extract of Caulis Spatholobi groups was also significantly down-regulated (p < 0.05). It showed that the ethanol extract of Caulis Spatholobus could promote apoptosis by up-regulating the expression of Bax and caspase-3 and down-regulating the expression of the Bcl-2 protein.
The treatment of Caulis Spatholobi for CML has the characteristics of multi-targets and multi-pathways. The results of in vitro pharmacological experiments demonstrated that its mechanism of action might be based on the expression of key target proteins, such as Caspase-3, Bcl-2, and Bax, thereby inhibiting cell proliferation and promoting cell apoptosis, which provides a scientific basis for the treatment of CML.
鸡血藤是血液病医师治疗恶性肿瘤的常用中药之一，但它的潜在靶点和分子机制仍需要进一步探索。
本研究旨在应用网络药理学方法预测鸡血藤治疗慢性髓系白血病（CML）的相关靶点，并通过体外细胞实验验证鸡血藤治疗 CML 的作用机制。
通过 TCMSP、ETCM、Genecards 和 GisGeNET 数据库获取鸡血藤治疗 CML 的相关靶点。利用 David 数据库进行 GO 和 KEGG 分析。使用 Cytoscape 3.7.2 构建“活性化合物-靶点-通路”网络。进一步通过体外药理学实验进行验证。采用 MTT 法和 Hoechst 33242 荧光染色法观察 K562 细胞的增殖和凋亡情况。采用 Western blot 验证预测靶点及其相关信号通路。
本研究共获得 18 个活性化合物和 43 个潜在靶点。MTT 法结果显示，与正常对照组相比，62.5-500μg/mL 浓度的鸡血藤醇提物对 K562 细胞均有明显的抑制作用，IC 值均小于 100μg/mL。Hoechst 33242 荧光染色法结果显示，鸡血藤醇提物能促进细胞凋亡。Western blot 结果显示，与正常对照组相比，62.5 和 125μg/mL 鸡血藤醇提物组 Bax 和 Caspase-3 蛋白表达明显上调（p<0.05）；125μg/mL 鸡血藤醇提物组 Bcl-2 蛋白表达明显下调（p<0.01），62.5 和 31.25μg/mL 鸡血藤醇提物组 Bcl-2 蛋白表达也明显下调（p<0.05）。说明鸡血藤醇提物可能通过上调 Bax 和 Caspase-3 的表达、下调 Bcl-2 蛋白的表达，从而促进细胞凋亡。
鸡血藤治疗 CML 具有多靶点、多通路的特点。体外药理实验结果表明，其作用机制可能基于 Caspase-3、Bcl-2、Bax 等关键靶蛋白的表达，从而抑制细胞增殖，促进细胞凋亡，为治疗 CML 提供了科学依据。
