Marsdenia tenacissima injection induces the apoptosis of prostate cancer by regulating the AKT/GSK3β/STAT3 signaling axis / 中国天然药物

Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKTSer473 and p-GSK3βSer9, and decreased the expression of p-STAT3Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.

Study on HPLC fingerprint of Myristica fragrans and its components identification / 国际中医中药杂志

Objective:To establish a method for quality evaluation of Myristica fragrans Houtt. Methods:The common peak was determined with Dehydroisoeugenol as the reference peak, and the HPLC fingerprint of Myristica fragrans was established; then the common peaks were analyzed by High Resolution Liquid Chromatography-mass Spectrometry (HPLC-MS). The chemical components of the common peaks were identified through the calculation and data retrieval of the primary and secondary mass spectra of the characteristic peaks. Results:The HPLC fingerprint of Myristica fragrans was established, and the similarity degree of the 10 batches of samples was above 0.9 and 11 common peaks were established. According to the results of HPLC-MS, the components of 11 common peaks were identified as follow: Methyl eugenol (peak 1), Licarin A (peak 2), Myristol (peak 3), OdoratisolA (peak 4), 2-(3,4-Dimethoxyphenyl) butynoic acid (peak 5), Malabaricon D (peak 6), 5'-Methoxydehydroisoeugenol (peak 7), Dehydroisoeugenol (peak 8), Malabaricone C (peak 9), 4-Methoxy-6-{(2S,3S)-7-methoxy-3-methyl-5-[(1E)-1-propen-1-yl]-2,3-dihydro-1- benzofuran-2-yl}-1,3-benzodioxole (peak 10) and Licarin B (peak 11). Conclusions:The quality of Myristica fragrans could be evaluate with HPLC fingerprint method. HPLC-MS was used to analyze the chemical composition of complex components, which will provide reference for the identification and analysis of chemical components of the extracts and preparations of Traditional Chinese Medicine.

Simultaneous Determination of 6 Flavonoids in Zhuang Medicine Engelhardia roxburghiana by HPLC-QAMS and Multivariate Statistical Analysis / 中国药房

OBJECTIVE：To establish a m ethod for simultaneous determination of neoastilbin ，astilbin，neoisoastilbin，isoastilbin， quercitrin and engeletin in Engelhardia roxburghiana，and conduct multivariate statistical analysis. METHODS ：HPLC-QAMS method was adopted. The determination was performed on Phenomenex SuperLu C 18 column with mobile phase consisted of acetonitrile-0.1％ formic acid （19 ∶ 81，V/V）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 254 nm （neoastilbin，astilbin，neoisoastilbin，isoastilbin，engeletin）and 291 nm（quercitrin）. The column temperature was 30 ℃，and sample size was 10 μL. Using astilbin as internal substance，and the relative correction factors of other 5 factors were calculated. The contents of each component were calculated according to relative correction factor ，and were compared with the results of external standard method. SPSS 22.0 software was used for cluster analysis and principal component analysis. RESULTS ：The linear range of neoastilbin ，astilbin，neoisoastilbin，isoastilbin，quercitrin and engeletin were 0.007-0.311，0.871-18.184，0.002-0.119， 0.052-1.251，0.105-2.202，0.020-2.319 μg（r＞0.999），respectively. RSDs of precision ，reproducibility and stability （24 h）tests were all lower than 3％. The average recoveries were 97.32％，94.89％，97.15％，96.90％，97.52％ and 97.53％（RSDs were 1.09％ -2.60％ ，n＝6），respectively. The relative correction factors of neoastilbin ，neoisoastilbin，isoastilbin，quercitrin and engeletin were 1.252 6，1.198 3，0.958 6，0.807 1 and 1.138 1， respectively. The contents of neoastilbin ， neoisoastilbin， qq.com isoastilbin，quercitrin and engeletin measured by QAMS were 0.394 2-2.067 2，0.139 1-0.804 7，2.864 8-8.554 8，4.581 2- 11.371 1，1.028 9-13.401 5 mg/g；the contents of neoastilbin ， astilbin，neoisoastilbin，isoastilbin，quercitrin and engeletin were 0.367 2-1.925 3，46.361 1-126.342 1，0.138 1-0.798 8，2.966 2-8.857 8， 4.642 5-11.523 3，0.970 6-12.641 9 mg/g，respectively. Relative errors of two methods was lower than or equal to 3.55％. The results of cluster analysis showed that 9 batches of samples could be clustered into two categories ；S8 sample was one category and others were one category. The results of principal component analysis showed that accumulative contribution rate of former 2 principle components was 84.745％，and the results of sample classification were consistent with those of cluster analysis. CONCLUSIONS ： The established HPLC-QAMS method is accurate ，feasible and repeatable ，and can be used for simultaneous determination of 6 flavonoids in E. roxburghiana ，and it can provide reference for quality control.

Clinical early intervention of Tongxia Huayu Decoction on pancreatic microcirculatory disturbance in severe acute pancreatitis / 中西医结合学报

OBJECTIVE: To study the mechanisms of Tongxia Huayu Decoction (a Chinese herbal decoction for purgation and removing blood stasis) in prognostic improvement for severe acute pancreatitis by early intervention on pancreatic microcirculatory disturbance. METHODS: Fifty-three patients with severe acute pancreatitis were divided randomly into treatment group (n=28) and control group (n=25). Tongxia Huayu Decoction was given to the patients in treatment group in addition to the normal treatment in control group for one week. The clinical symptoms and signs, hemodiastase, urinary amylase, C-reactive protein (CRP) and endothelin (ET) of the patients in the two groups before and after treatment were observed and detected. RESULTS: The total response rate of the treatment group was 98.4%, while that of the control group was 80%, with significant difference between them (P<0.05). There was no significant difference of the contents of hemodiastase, urinary amylase, CRP and ET between the two groups before treatment, while they were significantly decreased after treatment (P<0.01) with more obvious change in treatment group (P<0.01). CONCLUSION: Tongxia Huayu Decoction brings satisfied therapeutic effect on severe acute pancreatitis. The mechanisms may associate with its reducing function on ET releasing so as to inhibit the pancreatic microcirculatory disturbance and acinar cell injury induced by ET.