Comparison of pharmacokinetic behavior of glycyrrhizic acid and Bu-Zhong-Yi-Qi-Wan in rats / 国际药学研究杂志

Objective To establish a sensitive,simple and accurate HPLC-MS/MS method to quantify glycyrrhetic acid(glyc?yrrhetinic acid)in mice blood,and to further study pharmacokinetic profiles of glycyrrhetic acid after oral administration of glycyrrhi?zin and Bu-Zhong-Yi-Qi-Wan(BY). Methods Rats were intragastric administered of glycyrrhizin(glycyrrhizic acid,61.5 mg/kg) and BY extract(3 g/kg,with the same mole of glycyrrhizin moiety),respectively. Plasma samples were collected after administration and extracted with liquid-liquid extraction,then by separated by liquid chromatography on a C8 reversion phase chromatographic col?umn with gradient elution. Concentration of glycyrrhetic acid was detected by the validated HPLC-MS/MS. Non-compartmental pharma?cokinetic profiles were constructed using the software of Das 2.0 software(Shanghai,China),and the pharmacokinetic parameters were compared using unpaired Student′s t-test. Results This bioanalytical method was fully validated and showed good linearity(r>0.99),wide dynamic range(5-1000 ng/ml),and favorable accuracy and precision. Compared with the glycyrrhizin pure form group, BY significantly reduced the Cmax and AUC0-t of glycyrrhetic acid by 56%and 76%,respectively. Whereas no significant differences in Tmax,T1/2 and MRT were observed between the two groups. Conclusion The constituents in the BY prescription have significantly reduced the oral bioavailability of glycyrrhetic acid in rats than those in the glycyrrhizin pure form and the results indicate that some components in the BY have an inhibition effect on the absorption process of glycyrrhizin in the gut.

Hepatitis B virus-3p-siRNA inhibits hepatitis B virus replication and activates interferon-βexpression in mice / 中华传染病杂志

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.

Coronavirus membrane-associated papain-like proteases induce autophagy through interacting with Beclin1 to negatively regulate antiviral innate immunity

Autophagy plays important roles in modulating viral replication and antiviral immune response. Coronavirus infection is associated with the autophagic process, however, little is known about the mechanisms of autophagy induction and its contribution to coronavirus regulation of host innate responses. Here, we show that the membrane-associated papain-like protease PLP2 (PLP2-TM) of coronaviruses acts as a novel autophagy-inducing protein. Intriguingly, PLP2-TM induces incomplete autophagy process by increasing the accumulation of autophagosomes but blocking the fusion of autophagosomes with lysosomes. Furthermore, PLP2-TM interacts with the key autophagy regulators, LC3 and Beclin1, and promotes Beclin1 interaction with STING, the key regulator for antiviral IFN signaling. Finally, knockdown of Beclin1 partially reverses PLP2-TM's inhibitory effect on innate immunity which resulting in decreased coronavirus replication. These results suggested that coronavirus papain-like protease induces incomplete autophagy by interacting with Beclin1, which in turn modulates coronavirus replication and antiviral innate immunity.

SARS coronavirus papain-like protease inhibits the type I interferon signaling pathway through interaction with the STING-TRAF3-TBK1 complex

SARS coronavirus (SARS-CoV) develops an antagonistic mechanism by which to evade the antiviral activities of interferon (IFN). Previous studies suggested that SARS-CoV papain-like protease (PLpro) inhibits activation of the IRF3 pathway, which would normally elicit a robust IFN response, but the mechanism(s) used by SARS PLpro to inhibit activation of the IRF3 pathway is not fully known. In this study, we uncovered a novel mechanism that may explain how SARS PLpro efficiently inhibits activation of the IRF3 pathway. We found that expression of the membrane-anchored PLpro domain (PLpro-TM) from SARS-CoV inhibits STING/TBK1/IKKε-mediated activation of type I IFNs and disrupts the phosphorylation and dimerization of IRF3, which are activated by STING and TBK1. Meanwhile, we showed that PLpro-TM physically interacts with TRAF3, TBK1, IKKε, STING, and IRF3, the key components that assemble the STING-TRAF3-TBK1 complex for activation of IFN expression. However, the interaction between the components in STING-TRAF3-TBK1 complex is disrupted by PLpro-TM. Furthermore, SARS PLpro-TM reduces the levels of ubiquitinated forms of RIG-I, STING, TRAF3, TBK1, and IRF3 in the STING-TRAF3-TBK1 complex. These results collectively point to a new mechanism used by SARS-CoV through which PLpro negatively regulates IRF3 activation by interaction with STING-TRAF3-TBK1 complex, yielding a SARS-CoV countermeasure against host innate immunity.

Experimental study on anti-diarrhae effect of Gegenqinlian Decoction and its compounds / 中成药

AIM: To investigate combination principle of Gegenqinlian Decoction through experiment of anti-diarrhea. METHODS: Propelling rate of mouse's small intestine was selected as index on the condition of ink as colour reagent, diarrhea frequence served as second index for orthoganal design on the basis of mouse's diarrhea induced by senna. RESULTS:Coptis chinensis franch and Glycyrrhiza uralensis fisch and reduce drive rate. Scutellaria baicalensis Georgi and Pueraria lobata(wild) ohwi had counteracter against Coptis chinensis franch's effect, while Glycyrrhiza uralensis fisch could enforce Coptis chinensis franch's effect.as to DI, Coptis chinensis franch had anti-diarrhea effect. Scutellaria baicalensis Georgi had counteracter against Coptis chinensis franch's effect, Pueraria lobata(wild) Ohwi could enforce Scutellari, baicalensis Georgi effect, but Glycyrrhiza uralensis fisch could enforce Coptis chinensis's effect after compounding with Scutellaria baicalensis Georgi. CONCLUSION: ~Among the combinantion of Gegenginlian Decoction, as to the pharmcological effects on anti-diarrhea, the best compound is Coptis chinensis franch and Glycyrrhiza uralensis fisch.

Study on pharmacology of Wunzhonganwei Soft Capsule / 中成药

AIM: To evaluate the main pharmacological effect of Wunzhonganwei Soft Capsule. METHODS: Analgesic effect was investigated by hot plate test and writhing test, anti-inflammation effect by auricular swelling test in mice and toe swelling test in rat, anti-diarrhea effect by diarrhea test induced by rhubarb, effect on gastric emptying by phenolsulfonphthalein empty test in mice and effect on small intestine propulsive test by charcol powder propulsive rate test. RESULTS: Wunzhonganwei Soft Capsule enhanced thermal stimulation threshold in mice, decreased the occurrence of writhing caused by glacial acetic acid in mice, inhabited xylene-induced auricular swelling in mice and carrageenan-induced toe swelling in rat, decreased the number of loose stools induced by rhubarb in mice., inhabited the function of gastric emptying induced by metoclopramide or in normal mice, antagonized the inhibitory effect of gastric emptying induced by atropine and inhabited small intestine propulsive movement induced by neostigmineor or in normal mice. CONCLUSION: Wunzhonganwei Soft Capsule has the effect of anti-inflammation, analgesia, antidiarrhea and adjusting the function of gastrointestinal movement.

Effect of Xingshen Capsule on anti-centre and body fatigue in mice / 中成药

AIM: To investigate the action of Xingshen Capsule(Tea, Radix Ginseng, etc.) on anti-centre and body fatigue in mice. METHODS: Mice were divided into five groups and were given different dosage once a day for six days, then, their weight, motor activity, roating bar tolerance and the hypnogenetic effects of sodium pentobarbital were observed on the 1 st and 6 th day. RESULTS: Comparing with the negative control, Xingshen Capsule could notably increase the times of motor activity, prolong the persisting time on the roating bar, shorten the sleeping time caused by sodium pentobarbital while prolong its latence. CONCLUSION: Xingshen Capsule may have outstanding effect on refreshing one's nerve and body.