Research progress on the material basis and pharmacological effects of Pien Tze Huang / 药学学报

The medicinal history of Pien Tze Huang is long, and it is the only "double top secret" variety of technology and formula at present. It has the effects of clearing heat and detoxifying, detumescence and pain, cooling blood and removing blood stasis. At present, researchers have analyzed and identified some compounds in Pien Tze Huang and its precious medicinal materials, such as Panax notoginseng, calculus bovis, snake gall and musk, and conducted activity screening, pharmacokinetics and pharmacological related studies on these chemical components. It was found that Pien Tze Huang had a significant effect on the treatment of acute and chronic hepatitis, ulcer, colon cancer, liver cancer and other diseases. The purpose of this paper is to systematically discuss the research achievements of researchers in recent years on the material basis, pharmacological effects and clinical application of Pien Tze Huang, with a view to providing ideas for the further research of Pien Tze Huang.

Research progress on the pharmacological effects and chemical constituents of Pien Tze Huang and its potential Q-markers / 中国天然药物

Pien Tze Huang (PTH) was documented as an imperial prescription composed of Notoginseng Radix, Calculus Bovis, Snake Gallbladder, and Musk. It is famous in China and Asian countries due to its excellent effects in heat clearing, detoxifying, swelling reduction, and pain relieving. Modern pharmacological studies demonstrate that PTH shows excellent effects against various inflammatory diseases, liver diseases, and cancers. This review summaries the pharmacological effects, clinical applications, and mainchemical components of PTH. More importantly, its potential quality markers (Q-markers) were then analyzed based on the "five principles" of Q-markers under the guidance of Traditional Chinese Medicine theory, including transfer and traceability, specificity, efficacy, compatibility, and measurability. As a result, ginsenosides Rb1, ginsenoside Rg1, ginsenoside Rd, ginsenoside Re, notoginsenoside R1, dencichine, bilirubin, biliverdin, taurocholic acid, and muscone are considered as the Q-markers of PTH. These findings will provide guidance and assistance for the construction of a quality control system for PTH.

Effect of Pien Tze Huang against EV71 / 中国中药杂志

This study aims to investigate the inhibitory effect of Pien Tze Huang(PZH) on enterovirus 71(EV71). To be speci-fic, chemiluminescence method was adopted to evaluate the toxicity of PZH to African green monkey kidney(Vero) cells and human rhabdomyosarcoma(RD) cells, and cytopathic effect(CPE) method to assess the inhibition on EV71-GFP reporter virus and EV71 C4 wild-type virus. The results showed that PZH had low cytotoxicity to Vero cells and RD cells, with the half-maximal cytotoxic concentration(CC_(50)) of about 0.691 3-0.879 2 mg·mL~(-1) for the two. In addition, PZH can effectively inhibit the replication of EV71 within the non-cytotoxic concentration range, and dose-dependently alleviate the cytopathic changes caused by virus infection, with the half-maximal effective concentration(EC_(50)) of 0.009 2-0.106 3 mg·mL~(-1). On the basis of the above results, the green fluorescent protein(GFP), indirect immunofluorescence assay(IFA), and median tissue culture infective dose(TCID_(50)) were employed to assess and verify the anti-EV71-GFP and anti-EV71 C4 activity of PZH. The results demonstrated that PZH can dose-dependently lower the expression of GFP by EV71-GFP and structural protein VP-1 by EV71 C4 and decrease the production of progeny infectious viruses. The EC_(50) of PZH for EV71-GFP and EV71 C4 was about 0.006 0-0.006 2 mg·mL~(-1) and 0.006 6-0.025 6 mg·mL~(-1), respectively. This study suggested that PZH may exert antiviral activity by acting on EV71 and interfering with the expression of VP-1. At the moment, there is still a lack of specific anti-EV71 drugs. This study proposed a new idea for the symptomatic treatment of EV71 infections such as hand-foot-mouth disease and verified an effective drug for the treatment of EV71 infections.

Construction of Evaluating Method and Experimental Research for Targeting of Traditional Chinese Medicine Preparations Based on Supramolecular Qi Chromatography Theory / 中国实验方剂学杂志

Objective:To construct the targeting evaluation method of traditional Chinese medicine （TCM） preparations based on supramolecular Qi chromatography theory， and to study the liver targeting effect of Bupleuri Radix on Pien Tze Huang. Method:The molecular connectivity index （MCI） was used to analyze the characteristics of imprinted template and liver targeting tendency of TCM mainly attributed to liver meridian and components of Pien Tze Huang， and combined with target dynamics and total statistical moment principle， aimed at the independent action characteristics of multi-component imprinted template of TCM， a method for evaluating the targeting of TCM preparations was established. Hepatoma rats in Pien Tze Huang group， Bupleuri Radix<italic> </italic>group， Pien Tze Huang+Bupleuri Radix group and blank group were tested and verified. Result:After the average value of MCI of TCM mainly attributed to liver meridian was deducted， the MCI similarity between Pien Tze Huang group and Bupleuri Radix group was 0.376 8， Pien Tze Huang+Bupleuri Radix group and Bupleuri Radix group was 0.988 2， so it was predicted that Bupleuri Radix could enhance the liver targeting of Pien Tze Huang. A system for evaluating the targeting of TCM compounds was established， including relative total uptake efficiency （RUE_T）， relative total concentration （RC_T）， relative imprinted tendency （RIT_T） and relative imprinted variance （RIV_T）. The RUE_T and RC_T of liver were the highest in all tissues （RUE_T=1.88>1，RC_T<italic>=</italic>2.30>1）， and the corresponding values of other tissues were all <1， indicating that Pien Tze Huang combined with Bupleuri Radix could increase its distribution in liver and enhance liver targeting. Except for plasma， the RIT_T and RIV_T of other tissues fluctuated around 1.0， indicating that targeted modification did not change imprinted tendency of Pien Tze Huang and had no significant effect on the types of components. Conclusion:Under the guidance of supramolecular Qi chromatography theory， a targeting evaluation parameter system can be established to characterize the multi-component imprinted effect of TCM preparations by MCI and total statistical moment parameters， so as to realize the evaluation of targeting of TCM preparations. The addition of Bupleuri Radix can increase the liver targeting of Pien Tze Huang.

Exploring Anti-inflammation Mechanism of Pien Tze Huang via Regulating of Microglia Polarization / 中国实验方剂学杂志

Objective::To investigate the anti-inflammation mechanism of Pien Tze Huang (PTH) via regulating microglia polarization. Method::The experiment was divided into five groups, Blank, M1[lipopolysaccharide (LPS) 100 μg·L-1+ interferon-γ(IFN-γ) 10 μg·L-1], M1-PTH group[LPS 100 μg·L-1+ IFN-γ 10 μg·L-1+ PTH 0.4 g·kg-1], M2 group[interleukin-4 (IL-4) 20 μg·L-1], and M2-PTH group[IL-4 20 μg·L-1+ PTH 0.4 g·kg-1]. The concentration of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and transforming growth factor-β1(TGF-β1) in the culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA), the levels of inducible nitric oxide synthase(iNOS) and arginine-1 (Arg-1) mRNA were detected by real-time fluorescent quantitative polymerase chain reaction technique(Real-time PCR), and the expression levels of p-STAT1, p-STAT3, iNOS, p-STAT6, and Arg-1 were detected by Western blot. Result::The concentration of NO and TNF-α of the culture supernatant, the level of iNOS mRNA, as well as the level of p-STAT1, p-STAT3 and iNOS in M1 group, which were significantly increased(P<0.01) .Compared with blank group, but the concentration of NO and TNF-α were down-regulated(P<0.01), and iNOS mRNA(P<0.05), as well as the expression of iNOS, p-STAT1, and p-STAT3 was decreased (P<0.05, P<0.01) after the invention of PTH in M1-PTH group compared with M1 group. The concentration of IL-10 and TGF-β1 in the culture supernatant, the mRNA level of Arg-1, as well as the levels of p-STAT6 and Arg-1 were significantly increased in M2 group when compared with Blank group, addition to the concentration of IL-10 and TGF-β1 were up-regulated(P<0.05, P<0.01), and the expression of Arg-1 mRNA, the level of Arg-1, p-STAT6 were enhanced(P<0.05, P<0.01) in M2-PTH group compared with M2 group. Conclusion::PTH plays an anti-inflammatory role via regulating microglia polarization.

Inhibitory effect and mechanism of Pien Tze Huang on proliferation, migration, and invasion of lung cancer cells / 中草药

Objective: To study the inhibitory effect of Pien Tze Huang on proliferation, migration, and invasion of lung cancer cells and explore its mechanism. Methods: Human lung cancer cell A549 was used as the experimental object in vitro, and the cells were treated with different concentrations of Pien Tze Huang. MTT was used to detect cell proliferation, and the changes of cell invasion and migration were measured by Transwell assay. Western blot was used to detect the phosphorylation of invasive migration proteins MMP-9, MMP-2 and EMT proteins E-cadherin, vimentin and Akt signaling pathway proteins. Lung cancer cells were treated with Akt signaling pathway activator and tablets to detect the effect of activating Akt signaling pathway on the proliferation, invasion, and migration of lung cancer cells. Results: Pien Tze Huang (100 μg/mL tablets) had no effect on the proliferation of lung cancer cells, the proliferative ability of lung cancer cells could be significantly reduced by 200 μg/mL Pien Tze Huang. The invasion and migration ability of lung cancer cells treated with 100 and 200 μg/mL tablets were decreased, the protein expressions of MMP-2 and MMP-9 in cells were decreased, the level of E-cadherin protein in cells was increased, the level of vimentin protein was decreased, at the same time, the phosphorylation level of Akt was decreased. Akt signaling pathway activator treatment could significantly reduce the inhibitory effect of Pien Tze Huang on proliferation, invasion and migration of lung cancer cells (P < 0.05). Conclusion: Pien Tze Huang decreased the proliferation, migration and invasion of lung cancer cells, the mechanism is related to the inhibition of the activation of Akt signaling pathway.

Pien Tze Huang () Overcomes Doxorubicin Resistance and Inhibits Epithelial-Mesenchymal Transition in MCF-7/ADR Cells / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of Pien Tze Huang (, PZH) on breast cancer chemoresistance and related epithelial-mesenchymal transition (EMT) and investigate the underlying mechanisms.@*METHODS@#3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to determine the cell viability. Adriamycin (ADR) staining observed by fluorescence microscope was performed to detect the accumulation of ADR. Transwell assay was used to analyze the cell migration and invasion. Western-blot was performed to detect the protein expression of related genes.@*RESULTS@#MCF-7/ADR cells were resistant to ADR treatment, and PZH treatment inhibited the viability of MCF-7/ADR cells in a dose-dependent manner. PZH treatment also increased the intercellular accumulation of ADR and down-regulated the expression of ABCG2 and ABCB1 in MCF-7/ADR cells (P<0.05). In addition, PZH treatment inhibited EMT, migration and invasion of MCF-7/ADR cells (P<0.05). Moreover, PZH suppressed activation of transforming growth factor β1 (TGF-β) signaling in MCF-7/ADR cells (P<0.05).@*CONCLUSION@#PZH treatment can effectively overcome chemoresistance via down regulating ABCG2, ABCB1 and inhibit EMT in ADR resistant human breast cancer cells via suppression of the TGF-β1 pathway.

Determination of saponins in Pien Tze Huang by near infrared spectroscopy / 中国中药杂志

Near infrared spectroscopy(NIRS) was used for rapid quantitative analysis of saponins in Pien Tze Huang troches and powders. The near infrared spectra of Pien Tze Huang were collected,and the contents of notoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 in Pien Tze Huang were determined by high performance liquid chromatography(HPLC) as the reference values. Then the near infrared spectra of the samples were associated with the reference values to establish the quantitative analysis models by using partial least squares(PLS) method. Finally,the models were verified by unknown samples. The results showed that root mean square error of cross-validation(RMSECV) of R1,Rg1,Rb1 and the total content was 0.095 1,0.555,0.414,0.960 mg·g-1 for the troches models,0.085 6,0.443,0.405,0.913 mg·g-1 for the powders models. After external validations,root mean square error of prediction(RMSEP) of R1,Rg1,Rb1 and the total content was 0.111,0.274,0.276,0.807 mg·g-1 for the troches models,0. 059 2,0. 322,0. 327,0. 705 mg·g-1 for the powders models. The averages of relative standard deviation between the predicted values and the chemical measured values were all less than 2.0%. According to the results of paired-t tests at the level of α = 0.05,there were no significant differences between the predicted values and the measured values. The established quantitative analysis models can be used to predict the contents of saponins in Pien Tze Huang accurately and the proposed method is simple,fast,non-destructive and environmentally friendly for the rapid detection and quality control of saponins in Pien Tze Huang.

Effects of pien-tze-huang on expression of NMDAR1 and GluR2 in cortex of MCAO rats / 中国药理学通报

Aim: To investigate the effects of Pien-Tze-Huang (PZH) on mRNA and protein expression of NMDAR1 and GluR2 in cortex of MCAO rats. Methods: Forty healthy adult male Sprague-Dawley rats were randomly divided into three groups: sham, MCAO, MCAO + PZH groups. The rats were subjected to focal cerebral ischemia/reperfusion with suture-occluded method. Neurological deficit testing was performed with Zea Longa scale. The volume of cerebral infarction was evaluated by TTC staining. The mRNA and protein expression of NMDAR1 and GluR2 in cortex of side cerebral ischemic tissues were determined using qPCR and Western blot analysis. Results: Compared with MCAO group, PZH significantly improved the neurological deficit, decreased the volume of cerebral infarction, and up-regulated the mRNA and protein expression of NMDAR1 and GluR2. Conclusions: PZH attenuates the down-regulation of mRNA and protein expression of NMDAR1 and GluR2 after focal cerebral ischemic injury in rats, which may be associated with the cerebral protective effect of PZH.

Pien Tze Huang promotes HepG2 apoptosis via regulating ANXA1/VEGF signaling pathway / 中国药理学通报

Aim To explore the effect of Pien Tze Huang (PZH) on inhibiting HepG2 cells via regulating ANXA1/VEGF signaling pathway and the underlying mechanism. Methods HepG2 cells were treated with different concentrations (1, 1 0, 100 mg • L ~ l ) of PZH. MTT assay and colony formation assay were used to calculate cell viability and cell survival. Western blot was used to determine the expression of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, and ANXA1/VEGF signaling pathway protein, such as ANXA1, VEGF, VEGFR, NF-KB P 5 0 . Results Compared with normal group, different concentrations of PZH inhibited HepG2 cells in a dose-dependent manner, inhibited the colony formation, promoted the expression of apoptotic expression, promoted the expression of ANXA1 protein, inhibited the expression of VEGF, VEGFR, and N F - K B P 5 0 as well. Conclusions PZH can inhibit the activity of HepG2 cells in vitro. Its main mechanism is related to the promotion of apoptotic protein in HepG2 cells, the promotion of cell ANXA1 protein, and the inhibition of VEGF/VEGF receptor and NF-KB signaling pathway.

Pien Tze Huang Inhibits the Biological Behavior of Tongue Squamous Carcinoma Tca8113 Cells / 中国医科大学学报

Objective To investigate the effects of Pien Tze Huang on the biological behavior of tongue squamous carcinoma Tca8113 cells and its molecular mechanism.Methods Tca8113 cells were incubated with Pien Tze Huang at different doses.The proliferation of Tca8113 cells was examined by MTT assay.The distribution of the cell cycle was evaluated by PI staining.The expression of the p-STAT3 and Bcl-2 proteins was assessed by Western blotting.Results MTT assay showed that Pien Tze Huang inhibited the proliferation of Tca8113 cells in a dose-dependent manner.PI staining showed that Pien Tze Huang arrested cells in the G0/G1 phase.The results of Western blotting suggested that Pien Tze Huang inhibited STAT3 protein phosphorylation and Bcl-2 protein expression.Conclusion Pien Tze Huang inhibits the proliferation of tongue squamous carcinoma cells.

Inhibitory effect of human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang on human osteosarcoma transplant mice model / 中华中医药杂志

Objectives: To study the e ect of human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang on human osteosarcoma mice model. Methods: 40 standardized human osteosarcoma mice model were divided into 5 groups, treated with PBS, Paav-MCS, human P27KIP1 gene AVV virus, Chinese herb Pien Tze Huang or human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang in each di erent group. Results: ① Mice model of transplant human osteosarcoma were established successfully. ②Osteosarcoma were inhibited successfully by human P27KIP1 gene AVV virus, Chinese herb Pien Tze Huang or human P27KIP1 gene AVV virus combining with Chinese herb Pien Tze Huang. ③Mice model of transplant human osteosarcoma were active during observation. Conclusions: ①These results suggested that human P27KIP1 gene AVV virus, herb Pien Tze Huang and human P27KIP1 gene AVV virus combined with herb Pien Tze Huang were e ective on human osteosarcoma. ②Among them, the most e ective treatment was human P27KIP1 gene AVV virus combining with herb Pien Tze Huang. ③Chinese herb Pien Tze Huang was possible useful for life improvement of osteosarcoma patient.