Amino acid metabolism characteristics of Banxia Baizhu Tianma Decoction in realizing drug withdrawal based on transcriptomic analysis / 中国中药杂志

This study aimed to demonstrate the effect of Banxia Baizhu Tianma Decoction(BBTD) on realizing withdrawal of anti-epileptic drugs and explore the relationship between BBTD and the amino acid metabolism by transcriptomic analysis in the rat model of epilepsy induced by lithium chloride-pilocarpine. The rats with epilepsy were divided into a control group(Ctrl), an epilepsy group(Ep), a BBTD & antiepileptic drug integrative group(BADIG), and an antiepileptic drug withdrawal group(ADWG). The Ctrl and Ep were given ultrapure water by gavage for 12 weeks. The BADIG was given BBTD extract and carbamazepine solution by gavage for 12 weeks. The ADWG was given carbamazepine solution and BBTD extract by gavage for the former 6 weeks, and then only given BBTD extract for the latter 6 weeks. The therapeutic effect was evaluated by behavioral observation, electroencephalogram(EEG), and hippocampal neuronal morphological changes. High-throughput sequencing was used to obtain amino acid metabolism-related differen-tial genes in the hippocampus, and the mRNA expression in the hippocampus of each group was verified by real-time quantitative polymerase chain reaction(RT-qPCR). The hub genes were screened out through protein-protein interaction(PPI) network, and Gene Ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were performed. Two ceRNA networks, namely circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA, were constructed for ADWG vs BADIG. The experimental results showed that compared with those in Ep, rats in ADWG were significantly improved in the behavioral observation, EEG, and hippocampal neuronal impairment. Thirty-four amino acid metabolism-related differential genes were obtained by transcriptomic analysis, and the sequencing results were confirmed by RT-qPCR. Eight hub genes were obtained through PPI network, involving several biological processes, molecular functions, and signal pathways related to amino acid metabolism. Finally, the circRNA-miRNA-mRNA ternary transcription network of 17 circRNA, 5 miRNA, and 2 mRNA, and a lncRNA-miRNA-mRNA ternary network of 10 lncRNA, 5 miRNA, and 2 mRNA were constructed in ADWG vs BADIG. In conclusion, BBTD can effectively achieve the withdrawal of antiepileptic drugs, which may be related to the transcriptomic regulation of amino acid metabolism.

Mechanism of Banxia Baizhu Tianma decoction to prevent the development of prehypertension and excessive phlegm-dampness syndrome in rats based on metabonomics technology / 药学学报

The alterations of serum biological endogenous chemicals in rats with phlegm dampness accumulation syndrome of prehypertension (PHT) were interfered by Banxia Baizhu Tianma decoction (BBT), and the metabolic regulatory pathway of BBT was clarified using serum metabonomics analysis. To replicate the rat model of prehypertension phlegm dampness syndrome, blood pressure, behavioral markers, and serum biochemical markers of rats were collected. BBT's effectiveness in controlling blood pressure and blood lipids was assessed, and changes in endogenous small molecules in rat serum were determined using UPLC-Q-Orbitrap MS metabolic analysis. The results showed that BBT could regulate 9 metabolites, including arachidonic acid, cholic acid, glycodeoxycholic acid, N-adenosyltyrosine, arginine, lysophosphatidylethanolamine (20:0/0:00), lysophospholipid (P-18:0), lysophospholipid (18:0), lysophospholipid (22:5(7Z,10Z,13Z,16Z,19Z)). MetaboAnalyst was used to analyze the metabolic pathway. There were 7 metabolic pathways closely related to the change of blood pressure in rats, among which arachidonic acid metabolic pathway was the most critical. The metabolism difference foreign body in the model rats tends to return to the normal level, which provides a research basis for the mechanism of BBT from the perspective of metabonomics. This study was approved by the Experimental Animal Welfare Ethics Review Committee of Shandong University of Traditional Chinese Medicine (approval number: SDUTCM20211103001).

Optimization of extraction technology of Banxia Baizhu Tianma Decoction by chemometrics combined with information entropy weighting / 中草药

Objective: The extraction process optimization method of chemometrics combined with information entropy weight was established and applied to the water extraction process optimization of Banxia Baizhu Tianma Decoction (BBTD), in order to fully ensure the effectiveness and quality consistency of classical prescription. Methods: Taking BBTD as the model drug, the fingerprint was established by HPLC method. The common peak area was analyzed by principal component analysis (PCA), the total factor score of PCA, the similarity of fingerprint and the yield of dry paste were used as evaluation indexes. L9(34) orthogonal design was used to investigate the effects of adding water, soaking time and boiling time on the extraction effect, and the Objective: weight of each index was determined by information entropy weighting method. The technological parameters of water extraction were optimized. Results: Twenty-six common peaks and seven compounds were identified by the similarity evaluation system of chromatographic fingerprints of traditional Chinese medicine. According to the results of comprehensive score, the optimum extraction process of the preparation was determined to be 12 times the amount of water, decoction twice, 1 hour each time. The average comprehensive score of the three batches of verification was 0.418 0 with RSD of 3.32%. Conclusion: The optimized process has high extraction rate, good stability and repeatability, and is suitable for the industrial production of BBTD.

Identification of quality markers (Q-marker) of Banxia Baizhu Tianma Decoction based on component difference analysis and multivariate statistical method / 中草药

Objective: To identify the quality markers (Q-marker) for Baizhu Tianma Decoction from different batches based on multivariate statistical analysis. Methods: The fingerprint data of 18 batches of Banxia Baizhu Tianma Decoction were processed by the Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine (2012A Edition). SPSS 20.0 was used for cluster analysis and principal component analysis (PCA). According to PCA results, cluster analysis was carried out to screen out the markers that might affect the quality difference of Banxia Baizhu Tianma Decoction and determine its content in the samples. Results: A total of 12 chromatographic peaks were selected as common peaks of fingerprint, and the similarity of 18 batches of samples was greater than 0.900, which indicated that each batch had good consistency. Eighteen batches of samples were clustered into three groups by cluster analysis. Five marker peaks were screened out from PCA results. Two of them were identified as gastrodin and naringin. The five marker peaks were further clustered and the results were basically the same as before. There was a certain correlation between the comprehensive score of principal components and the determination results of gastrodin and naringin in 18 batches of samples. Conclusion: Through multivariate statistical analysis, the Q-markers that may affect the difference of Banxia Baizhu Tianma Decoction were better identified, which can lay the foundation for the comprehensive quality control of Banxia Baizhu Tianma Decoction and its compound preparations, and provide reference for the further study of the correlation between the different components and pharmacodynamics of Banxia Baizhu Tianma Decoction.

Effects of Banxia Baizhu Tianma Decoction on Expression of miRNA-146a-5p in Hippocampus of Epileptic Rats and Bioinformatic Analysis / 中国中医药信息杂志

Objective To lay the foundation for studying the possible pathogenesis of epilepsy and the anti-epileptic mechanism of Banxia Baizhu Tianma Decoction through the bioinformatic analysis of target gene prediction and signal pathway of miRNA-146a-5p in hippocampus of epileptic rats. Methods Lithium-pilocarpine was used to induce seizures in rat models. The experiment rats were randomly divided into normal control group, model group, Banxia Baizhu Tianma Decoction group, with 20 rats in each group. The method of miRNA expression profiling was used to observe the miRNA differential expression of hippocampus neuron cell of rats. The expression level of miRNA-146a-5p was detected by real-time quantitative PCR. MiRDB was used for target gene prediction of miRNA-146a-5p, and miRTarBase and DAVID were used for enrichment analysis on the GO function and KEGG signaling pathway. Results The attack times and grades of the rats in Banxia Baizhu Tianma Decoction group were significantly lower than those in the model group from behavioral observation. MiRNA microarray analysis showed that the expression level of miRNA-146a-5p in model group was 2.107 times normal control group (P＜0.05), and the expression level decreased to 1.377 times after treatment with Banxia Baizhu Tianma Decoction (P＜0.05). The results of RT-PCR was consistent with that of miRNA microarray, with statistical significance (P＜0.05). MiRNA-146a-5p target gene prediction results had 140 target genes by GO, and there were 14 annotation information of biological process (P＜0.05), 9 annotation information of cellular component (P＜0.05), 11 annotation information of molecular function (P＜0.05). Enrichment analysis of KEGG biological pathway showed that 140 target genes of miRNA-146a-5p were enriched in EB virus infection signal pathway and thyroid hormone signaling pathway (P＜0.05). Conclusion miRNA-146a-5p is closely related to the inflammatory reaction after epilepsy, and Banxia Baizhu Tianma Decoction can control epilepsy possibly by controlling the inflammatory reaction after epilepsy.

Decomposition and Reformulation of Banxia Baizhu Tianma Decoction: A Vasodilatory Approach / 中草药·英文版

Objective To decompose and reformulate Banxia Baizhu Tianma Decoction (BBT) in order to optimize its ratio for vasodilatory effect. The fingerprints of these six component herbs were also studied via Tri-step FTIR analysis. Methods The herbal components of BBT were extracted separately with different solvents, such as 95% ethanol, 50% ethanol, and distilled water. The vasodilatory effect of the extracts was studied using the in vitro pre-contracted aortic ring model. Among these extracts, the water extracts of Pinellia ternata [PT(W)] and Citri reticulatae [CR(W)], 50% ethanol extracts of Gastrodia elata [GE(50)] and Glycyrrhiza uralensis [GU(50)], and 95% ethanol extracts of Atractylodes macrocephala [AM(E)], and Poria cocos [PC(E)] were selected for the orthogonal stimulus-response compatibility group study due to their high effectiveness in vasodilation. Results The best combination ratio (G2) was GE(50), GU(50), AM(E), PC(E), CR(W), and PT(W) at EC0, EC5, EC5, EC5, EC5, and EC5, with EC50 of (0.06 ± 0.01) mg/mL. Comparison between the vasodilatory effects of extracts derived from G2 and BBT were also studied. From the results, the 50% ethanol extract following G2 ratio was found to be most potent extract with the highest vasodilatory effect compared with other extracts derived from G2 or BBT, with EC50 of (0.05 ± 0.005) mg/mL. Conclusion The optimum ratio and appropriate extraction method for the new BBT formulae are obtained. G2 is determined as the best formulae with the highest potency in producing the vasodilatory effect on isolated rat aortic rings. The results obtained suggest that 50% ethanol is found to be the best solvent to extract the vasoactive compounds of G2 in order to exhibit the highest efficacy of vasodilatory effect.

Banxia-Baizhu-Tianma decoction combined with simvastatin for hyperlipidemia / 国际中医中药杂志

Objective To investigate therapeutic effect of the Banxia-Baizhu-Tianma decoction combined with simvastatin for phlegm and blood stasis obstructing hyperlipidemia. Methods A total of 120 patients with phlegm and blood stasis obstructing hyperlipidemia from Dalian central hospital from June 2010 to December 2013 were divided into 2 groups according to random digits table:60 patients in a control group treated with simvastatin 20 mg/d,and 60 patients in a treatment group treated with Banxia-Baizhu-Tianma decoction on the basis of the control group. After 6 weeks treatments, the clinical efficacy, serum levels of total cholesterol(TC), triglyceride(TG), high density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C), high sensitive C reactive protein(hs-CRP),high mobility group protein B1(HMGB1), alanine aminotransferase(ALT), γ-glutamyl transpeptidase(γ-GT), interleukin 8(IL-8)and superoxide dismutase(SOD) were evaluated in both groups. Results The total effective rate was 91.7%(55/60) and 73.3%(44/60) in the treatment group and the control group, respectively, with significant difference (χ2=5.772, P<0.05). After treatment, the serum levels of TC, TG, HDL-C, hs-CRP, HMGB1 in both groups were significantly lower than those before the treatment (in the treatment group t value were 12.355, 5.984, 32.825, 13.806, 14.661 and in the control group t value were 7.553, 2.177, 3.064, 10.281, 9.019, P<0.05 or P<0.01), LDL-C concentration significantly increased (t in the treatment group=3.382, P<0.01;t in the control group =2.388, P<0.05). After the treatment, the serum levels of TC, TG, HDL-C, hs-CRP, and HMGB1 in the treatment group were lower than those in the control group (t value were 7.173, 3.418, 2.377, 3.967, 8.352, P<0.05 or P<0.01);there was no significant difference in LDL-C concentration between the two groups (t=1.630, P=0.106);the levels of ALT, GT, IL-8 and SOD in the treatment group showed significant difference compared with those before the the treatment (t value were 19.053, 9.592, 12.909, 10.377, P<0.01), while in the control group, exceptγ-GT, other parameters showed significant difference after the treatment compared with before the treatment (t value were 2.253, 5.876, 3.266, P<0.05). Compared with the control group, these parameters in the treatment group were significantly different after the treatment(t value were 27.872, 10.244, 6.954, 6.145, P<0.01). Conclusions Banxia-Baizhu-Tianma decoction combined with simvastatin has good lipid-lowering effect. It may significantly lower inflammatory factor levels, reduce inflammation, and enhance antioxidant capacity.

Comparison of serum fingerprint in extract solution from Banxia Baizhu Tianma Decoction / 中草药

Objective Comparing the serum fingerprint of four kinds of extract solution from Banxia Baizhu Tianma Decoction.Methods Taking rabbits as experiment animals,collecting blood,and separating serum after drenching semi-bionic enzyme extracting(SBEE) solution.HPLC was used to detect the fingerprints of medicated serum at different blood-collecting time in order to optimize the best blood collecting time,then the serum fingerprints of four kinds of solution collected were compared.Results HPLC showed that when blood collecting time was 2 h,the number and total area of chromatographic peak had the maximum value;SBEE Solution had the the maximum number and total area of chromatographic peak in the four kinds of solution at the best blood collecting time.Conclusion The component content in SBEE medicated serum is higher than that by other method.