Progress in pharmacodynamic basis, mechanism, and prediction of Q-markers of Zhenwu Decoction in treatment of chronic heart failure / 中国中药杂志

Zhenwu Decoction(ZWD) has a history of more than 1 800 years in traditional Chinese medicine(TCM), which is used to treat various diseases characterized by Yangqi deficiency and exuberant water and dampness. It is currently the classic prescription for the treatment of chronic heart failure(CHF). This study provides a basis for the treatment of CHF with ZWD by elaborating the traditional efficacy, theoretical basis, and underlying mechanism of the prescription. Based on the research methods and judgment basis of quality markers(Q-markers) of Chinese medicine, the Q-markers of ZWD in the treatment of CHF were predicted from the aspects of transfer and traceability, specificity, effectiveness, compatibility environment, measurability, and processing. Demethyl-coclaurine,benzoylaconine, atractylenolide Ⅲ, paeoniflorin, 6-gingerol, 8-gingerol, pachymic acid, and dehydrotumulosic acid can be used as Q-markers of ZWD for treating CHF. The result provides a reference for exploring the pharmacodynamic substances of ZWD in the treatment of CHF.

Purification and characterization of Hainantoxin-V, a tetrodotoxin-sensitive sodium channel inhibitor from the venom of the spider Selenocosmia hainana.

A neurotoxic peptide, named Hainantoxin-V (HNTX-V), was isolated from the venom of the Chinese bird spider Selenocosmia hainana. The complete amino acid sequence of HNTX-V has been determined by Edman degradation and found to contain 35 amino acid residues with three disulfide bonds. Under whole-cell patch-clamp mode, HNTX-V was proved to inhibit the tetrodotoxin-sensitive (TTX-S) sodium currents while it had no any effects on tetrodotoxin-resistant (TTX-R) sodium currents on adult rat dorsal root ganglion neurons. The inhibition of TTX-S sodium currents by HNTX-V was tested to be concentrate-dependent with the IC(50) value of 42.3nM. It did not affect the activation and inactivation kinetics of currents and did not have the effect on the active threshold of sodium channels and the voltage of peak inward currents. However, 100nM HNTX-V caused a 7.7mV hyperpolarizing shift in the voltage midpoint of steady-state sodium channel inactivation. The results indicated that HNTX-V inhibited mammalian voltage-gated sodium channels through a novel mechanism distinct from other spider toxins such as delta-ACTXs, micro -agatoxins I-VI which bind to receptor site three to slow the inactivation kinetics of sodium currents.

Inhibition of sodium channels in rat dorsal root ganglion neurons by Hainantoxin-IV, a novel spider toxin.

The effects of Hainantoxin-IV (HNTX-IV), a neurotoxic peptide isolated from the venom of the Chinese bird spider Seleconosmia hainana, on the adult rat dorsal root ganglion (DRG) neurons were investigated. Using the whole-cell patch-clamp technique HNTX-IV inhibited mammal neural TTX-sensitive (TTX-S) sodium currents evidently but the toxin failed to affect TTX-resistant (TTX-R) ones. The inhibition of HNTX-IV is dose-dependent with the IC(50) value of 44.6 nmol/L. The toxin didn't affect the activation and inactivation kinetics of sodium currents, but it caused a 10.1 mV hyperpolarizing shift in the voltage midpoint of steady-state sodium channel inactivation on DRG neurons. The results indicated that HNTX-IV, a novel spider toxin, maybe alternate voltage-gated sodium channels through a mechanism distinct from other spider toxins such as delta-ACTXs, mu-agatoxins I-VI which targeted the receptor site 3 to slow the inactivation kinetics of sodium currents.

The significance of CD14 and TLR4 expressions in severe hepatitis B induced by endotoxin / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To explore the roles of CD14 and TLR4 in severe hepatitis B induced by endotoxin.</p><p><b>METHODS</b>The levels of mCD14 on PBMCs from 30 cases of severe hepatitis B, 20 cases of chronic hepatitis B and 20 cases of healthy controls were detected by flow cytometry. The expressions of CD14 mRNA and TLR4 mRNA in PBMCs from these patients were also detected by RT-PCR. Meanwhile, the concentration of plasma endotoxin was detected by limulus amebocyte lysate test and the levels of TNF alpha, IL-1, IL-6 in serum were detected by ELISA.</p><p><b>RESULTS</b>The levels of mCD14 on PBMCs in severe hepatitis B, chronic hepatitis B and control were 74.2+/-12.3, 63.6+/-11.8 and 60.3+/-7.2 respectively. There was a significant difference among severe hepatitis B,chronic hepatitis B and healthy controls (both of P less than 0.01). The expressions of CD14 mRNA and TLR4 mRNA (2.92+/-0.67 and 1.86+/-0.45) were also upregulated, compared with that in chronic hepatitis B patients (1.34+/-0.51, 0.93+/-0.18) and healthy group (0.92+/-0.58, 0.73+/-0.16) (P less than 0.01). Similarly, the concentration of plasma endotoxin was much higher in severe hepatitis B (1.87+/-1.61) than that in chronic hepatitis B patients (0.11+/-0.11) and that in healthy group (0.03+/-0.03) (P less than 0.01). As a result, the inflammation cytokines, such as TNF alpha, IL-1 and IL-6 (19.78+/-9.21, 0.96+/-0.16, 68.34+/-48.30) also increased significantly in severe hepatitis B, which were remarkably higher than those in chronic hepatitis B patients (7.26+/-6.52, 0.19+/-0.02 and 19.28+/-4.65) and healthy group (4.15+/-4.06, 0.15+/-0.01 and 12.01+/-3.88) (P less than 0.01). Furthermore, correlation analysis showed there was positive correlation among the level of mCD14, the expression of CD14 mRNA/TLR4 mRNA, the concentration of endotoxin and the levels of inflammation cytokines in severe hepatitis B (r1 = 0.865, r2 = 0.415, r3 = 0.524, all of P less than 0.05).</p><p><b>CONCLUSION</b>Endotoxin is an important factor in the aggravation of hepatitis B. Meanwhile, mCD14, CD14 mRNA and TLR4 mRNA are remarkably upregulated during the endotoxin stimulation. The inflammation cytokines (TNF alpha, IL-1 and IL-6) are also elevated, which may finally result in the aggravation of the hepatitis B. Therefore, CD14, TLR4 and inflammation cytokines play important roles in pathogenesis of severe hepatitis B induced by endotoxin.</p>

[Solid-phase synthesis and biological characterization of S12A-HNTX-IV and R29A-HNTX-IV: two mutants of hainantoxin-IV].

Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.