Chemicalome and metabolome profiling of Chai-Gui Decoction using an integrated strategy based on UHPLC-Q-TOF-MS/MS analysis.

Traditional Chinese medicine prescriptions are widely believed to exert therapeutic benefits via a multiple-component and multiple-target mode. The systemic profiling of their in vitro chemicalome and in vivo metabolome is of great importance for further understanding their clinical value. Herein, an integrated strategy using ultra-performance liquid chromatography coupled with quadruple time-of-flight mass spectrometry was proposed to profile the chemicalome and metabolome of Chai-Gui Decoction. Particularly, an approach combined mass defect filter, characteristic product ion filter, and neutral loss filter was adopted to identify metabolites in plasma, urine, bile, and feces by MetabolitePilot. Consequently, a total of 174 constituents were identified or tentatively characterized and 70 metabolites that related to 21 representative structural components were matched in rat biofluids. Among them, 19 prototypes and 7 metabolites that contributed to flavonoids, monoterpenes, and phenylpropanoids were detected distribution in brain, heart, kidney, liver, lung or spleen. This study provided a generally applicable approach to comprehensive investigation on chemicalome and metabolome of traditional Chinese medicine prescriptions, and offered reasonable guidelines for further screening of quality control indicators of Chai-Gui Decoction.

Effect of Shenqi Yangxin decoction on high mobility group box 1 and inflammatory signal pathway in a rat model of dilated cardiomyopathy.

OBJECTIVE: To investigate the effects of Shenqi Yangxin decoction (SQYXD) on heart function in a rat model of dilated cardiomyopathy (DCM) and its potential mechanisms. METHODS: Sprague-Dawley rats were randomly divided into normal (10 rats) and DCM (150 rats) groups. DCM was induced by an intraperitoneal injection of adriamycin. Then, DCM baseline group was randomly selected sixteen DCM rats. The remaining DCM rats were randomly divided into DCM control, perindopril, metoprolol, and SQYXD groups. Cardiac function and histological analysis plus biochemical measurement of serum levels of brain natriuretic peptide (BNP), and inflammatory factors were measured. The mRNA and protein expression levels of high-mobility group box 1 (HMGB1), Toll-like receptor 4 (TLR-4), receptor for advanced glycation end products (RAGE), and nuclear factor-¦ÊB (NF-¦ÊB) were determined. Myocardial metabolism imaging was performed on the normal, SQYXD and DCM control groups to evaluate the effectiveness of treatments. RESULTS: Rats in the DCM control group exhibited dilated left ventricular diameter, impaired cardiac function, disorganized sarcomere, impaired glucose metabolism, increased heart weight index, and increased levels of BNP, which were improved by treatment with SQYXD. In addition, hearts from rats in the DCM baseline group exhibited significantly higher levels of HMGB1, TLR-4, RAGE, NF-κB, tumor necrosis factor-α, interleukin-1, interleukin-6, interleukin-10, compared with the normal group. Interestingly, the mRNA level of HMGB1 in the DCM baseline group was positively correlated with that of TLR-4, RAGE, NF-κB, BNP, and LVEDD, but negatively correlated with LVEF. SQYXD inhibited the upregulation of HMGB1 expression and its downstream inflammatory factors. CONCLUSION: Shenqi Yangxin decoction effectively reduced the dilated left ventricular diameter and improved heart function in dilated cardiomyopathy. The mechanisms underlying the action on DCM involve regulating the gene and protein expression of HMGB1 and its inflammatory signal pathways in the DCM rat model.

Effect of Yiqi Yangyin prescription in treatment of dilated cardiomyopathy: a Meta-analysis of randomized controlled trials.

OBJECTIVE: To evaluate the therapeutic effect of Yiqi Yangyin prescription for dilated cardiomyopathy (DCM). METHODS: Electronic databases were searched for relevant clinical randomized controlled trials of DCM treatment using Yiqi Yangyin prescription (Shengmai and Zhigancao decoction). Databases searched included PubMed, EMBASE, the Cochrane Library, China National Knowledge Infrastructure Database, Wanfang Database, China Science and Technology Journal Database, and Chinese Biological Medicine from January 1, 1985, to October 31, 2015. After literature screening and data extraction according to previously determined exclusion criteria, data were analyzed using RevMan 5.3 and Stata 12.0. Study heterogeneity was evaluated using the I 2 test and Cochran's Q test. Egger's test was used to detect publication bias. RESULTS: Nineteen trials involving 1024 participants were included in the Meta-analysis. Of these, 15 used Shengmai (treatment group: 421 cases; control group: 344 cases) and 4 used Zhigancao decoction (treatment group: 133 cases; control group: 126 cases). A Meta-analysis demonstrated that the total curative effect was significantly improved by combining Shengmai with conventional treatment [relative risk (RR) = 1.32, 95% confidence intervals (CI) (1.22, 1.43), P < 0.01]. Left ventricular ejection fraction [standard mean difference (SMD) = 1.13, 95% CI (0.55, 1.70), P < 0.01] and left ventricular end diastolic [SMD = -0.46, 95% CI (-0.70, -0.23), P < 0.01] were also improved. Adding Zhigancao decoction achieved the same effect [total efficiency of RR = 1.34, 95% CI (1.16, 1.54), P < 0.01]. CONCLUSION: Compared with conventional therapy, Yiqi Yangyin prescription can significantly improve the curative effect, increase left ventricular ejection fraction, and reduce left ventricular end diastolic. Thus, it can effectively improve heart function in patients with DCM.

[Preparation of three functional monoclonal antibodies against human FL molecule and analysis of their bioactivity].

AIM: To Prepare three functional monoclonal antibodies(mAbs) against human FL molecule and analyze their bioactivity. METHODS: The cell line L929-FL transfected with human FL gene was used as immunogen. The hybridomas secreting the antibodies against human FL were obtained by fusing splenoecytes from the immunized mice with murine myeloma cells(Sp2/0). Their subclasses were analyzed using fast-strip method. The monoclonal antibodies were produced in mouse peritoneal cavity and purified by Protein G affinity chromatography. The inhibitory effect of mAbs against FL on leukemia cell lines U937 and HL-60 was detected by MTT. The apoptosis of U937 and HL-60 cells stained by annexin-V/PI was determined by FCM. RESULTS: Three hybridomas named 3C2, 3C6 and 8D10 were successfully obtained, which secreted monoclonal antibodies against human FL molecule stably. Their subclasses were the mouse IgG2a with kappa light chains. The three monoclonal antibodies recognized the FL molecule on U937 and HL-60 cells that also coexpressed Flt3 molecule. When U937 and HL-60 cells were cultured in presence of 3C2, 3C6 and 8D10, their proliferation was reduced as compared to that in control in MTT assay(P < 0.05). The analysis of annexin-V/PI binding to U937 and HL-60 cells by FCM showed the mAbs had the apoptotogenic activity of the monoclonal antibodies against human FL molecule. CONCLUSION: 3C2, 3C6 and 8D10 are three funtional monoclonal antibodies against human FL molecule. They may be of some value in the study of the roles of FL/Flt3 interaction in leukemia pathogenesis.