Identification and verification of effective components of Huanghuai for dysfunctional uterine bleeding based on network pharmacology and molecular docking / 中草药·英文版

Objective: The Huanghuai (HH), which is made from the dried roots of Scutellaria baicalensis (Huangqin in Chinese) and the dried flowers and buds of Sophora japonica (Huaihua in Chinese), is a traditional Chinese formula used to treat dysfunctional uterine bleeding (DUB) (Benglou in Chinese) and proven to treat hemostasis effectively in our previous study. Network pharmacology and molecule docking were performed to study the underlying mechanism of Huanghuai (HH), and pharmacodynamic experiments were conducted to verify its curative effect. Methods: TCMSP, UniProt, GeneCards, STRING, DAVID databases, and Cytoscape 3.7.2 were utilized for the construction of a compound-target-pathway network. Docking the potential effective components with potential targets. The HPLC analysis of the potential effective components was performed. In vivo, the hot plate test model was used to study the analgesic activity, the egg white was used to study the swollen reaction in the sole in mice, and the hemostasis effect was studied by the capillary method, tail-breaking method and abortion uterus test. Results: The results showed that six compounds (acacetin, beta-sitosterol, wogonin, baicalein, kaempferol and quercetin) and four potential targets (PTGS2, AKT1, TP53 and TNF) in the compound-target-pathway network were the potential material basis for HH to treat DUB. It can be seen that the binding energy of the acacetin, wogonin, baicalein, beta-sitosterol, kaempferol and quercetin in HH docked with the receptor proteins PTGS2, AKT1, TP53, and TNF were far less than −5.0 kJ/mol, which means the molecules have low conformational energy, stable structure and high binding activity. And the result of HPLC analysis showed that acacetin, wogonin, baicalein, kaempferol and quercetin were the potential effective components of the hemostasis mechanism of HH, beta-sitosterol was removed due to low content. In vivo testing of the potential effective components, it revealed that the group of potential effective components identified by HPLC could increase the pain threshold, inhibit the swelling hind paws of mice induced by egg white, reduce the bleeding time and clotting time, reduce uterine bleeding, decrease the uterine weight, increase the content of Ca and ET-1, and reduce the content of NO in uterine homogenate tissue, and decrease of E

Optimization of extraction technology from compound Huanghuai based on coagulation activity and central composite design-response surface methodology / 中国中药杂志

To establish a method for the determination of coagulation activity in vitro by using fibrinogen plate method. The extraction route of compound Huanghuai was optimized by selecting thrombin as the reference substance. The comprehensive score of extract yield and the extraction transfer rate of baicalin and rutin was set as the dependent variable, with alcohol concentration, solvent volume and extraction time as the influence factors in central composite design for quadratic fitting, and the extraction process of compound Huanghuai was optimized by using response surface methodology. The results of thrombin concentration and precipitation zone area showed a good linear relationship in 0.4-16 U•mL⁻¹, r=0.997 5. The average recovery rate was 103.8% and the RSD was 4.7 %. The circle of precipitation area of compound Huanghuai combined extract was 38.81 mm², which was bigger than that of fractionated extract. The activity on the daily amount of compound Huanghuai extract was 84.28 U; and the optimum extraction technology was as follows: alcohol concentration 40%, extracted 3 times, the liquid-solid ratio 6∶1, and extraction time 2 h. The predicted value of comprehensive score was 94.26 and the measured value was 88.34, respectively, with a relative deviation of 6.28%. The coagulation activity of the intermediate obtained by optimal extraction process was better. So the method established in this paper was simple, fast and accurate for determination of coagulation activity of compound Huanhuai, which can be also used for the screening of follow-up process and quality control.

Quality Standard Improvement for Compound Huanghuai Tablets / 中国药师

Objective:To improve the quality standard for compound Huanghuai tablets. Methods: The main components of the preparation including Flos Sophorae Immaturus, Radix Scutellariae Baicalensis, Radix Paeoniae Rubra, Radix et Rhizoma Rhei Palma-ti, Rhizoma Imperatae, Caulis Spatholobi, Semen Coicis and Spica Prunellae Vulgaris were identified by TLC qualitatively. The content of rutin in Flos Sophorae Immaturus was determined by HPLC. A DIKMA? Spursil C18 (250 mm × 4. 6 mm, 5 μm) column was used with methanol-0. 2% phosphoric acid solution (45:55) as the mobile phase. The flow rate was 1. 0 ml·min-1 , the detection wave-length was set at 350 nm, the column temperature was maintained at 25℃, and the injection volume was 20μl. Results:The spots in TLC were clear without any interference. The calibration curve was linear within the range of 0. 0116-0. 1859 mg · ml-1 ( r =0. 9999) for rutin. The average recovery was 98. 93% (RSD=1. 14%,n=6). Conclusion: The improved quality standard is sim-ple, specific and reproducible, which can be used for the quality control of compound Huanghuai tablets.