Anemoside B4 inhibits SARS-CoV-2 replication in vitro and in vivo / 中草药·英文版

OBJECTIVE@#Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro.@*METHODS@#The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining.@*RESULTS@#The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism.@*CONCLUSION@#Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

Effects of anemoside B4 on the expression of apoptosis-associated proteins in the retina of diabetic rats / 中华老年医学杂志

Objective:To investigate the effects of anemoside B4 on apoptosis of retinal cells in diabetic rats.Methods:Sixty Sprague-Dawley rats were randomized into three groups: the normal control(control), diabetic rats(DM)and diabetic rats treated with Anemoside B4(B4)groups(n=20, each group). Rats in the DM and B4 groups were rendered diabetic with an intraperitoneal injection of streptozotocin(STZ, 60 mg/kg). After 3 days of successful modeling, rats in the B4 group were intraperitoneally injected with anemoside B4(5 mg/kg), twice/day, for 8 weeks, while rats in the control and DM groups were injected with an equivalent volume of normal saline.After 8 weeks of anemoside B4 and normal saline injection, rats were sacrificed and retinas were harvested for examination.Paraffin sections of retina were stained with the hematoxylin-eosin(H-E)method for morphological evaluation.Protein levels of Bax and Bcl-2 were detected by using Western blot.The expression of caspase-3 mRNA was detected with quantitative PCR.Results:H-E staining results showed the control group had intact retinal structure and clear morphological features, whereas disordered retinal structure, thinner layers, and sparse and disorganized cells were seen in the DM group.However, retinal structure and morphology were improved after treatment with anemoside B4.Compared with the control group, the protein expression of Bcl-2 was lower( t=57.81, P<0.01), the protein expression of Bax was higher( t=10.47, P<0.01), and the Bcl-2/Bax ratio was lower( t=23.98, P<0.01)in the DM group.Compared with the DM group, the protein expression of Bcl-2 was higher( t=41.07, P<0.01), the protein expression of Bax was lower( t=6.811, P<0.01), and the Bcl-2/Bax ratio was higher( t=14.70, P<0.01)in the B4 group.Caspase-3 mRNA expression was higher in the DM group than in the control group( t=7.916, P<0.01), but was lower in the B4 group compared with the DM group( t=6.221, P<0.01). Conclusions:Anemoside B4 can inhibit the apoptosis of retinal cells by up-regulating Bcl-2 expression and down-regulating Bax and caspase-3 expression in diabetic rats.

A New Triterpenoid Saponin from Aidi Injection / 中草药·英文版

ObjectiveTo investigate the chemical constituents from Aidi Injection.Methods The chemical constituents were isolated by chromatography on Sephadex LH-20 gel columns and reverse phase semi-preparative HPLC repeatedly.Their structures were identified by spectroscopic analysis (NMR and MS).ResultsTwenty-two compounds were isolated and identified to be 3-O-3',4'-diacetyl-β-D-xylopyranosyl-6-O-β-D-glucopyranosylcycloastragenol (1),astragaloside IV (2),astragaloside Ⅱ (3),astragaloside I (4),isoastragaloside I (5),acetylastragaloside I (6),ginsenosid Re (7),ginsenoside Rf (8),ginsenoside Rg1 (9),ginsenoside Rb3 (10),notoginsenoside R4 (11),ginsenoside Rb1 (12),ginsenoside Rc (13),ginsenoside Rb2 (14),ginsenoside Rd (15),lucyoside H (16),3-O-ββ-D-glucopyranosyl(l→4)-β-D-glucopyranosyl(l→3)-α-L-rhamnopyranosyl (1→2)-α-Larabinopyranosyl oleanolic acid 28-O-α-L-rhamnopyranosyl(l→4)-β-D-glucopyranosyl(1→6)-β-D-glucopyranoside (17),3-O-β-D-glucopyranosyl(1→3)-α-L-rhamnopyranosyl [β-D-glucopyranosyl-(l→4)]-(l→2)-αt-L-arabinopyranosyl oleanolic acid 28-O-α-L-arabinopyranosyl(1→4)-β-D-glucopyranosyl(1→6)-β-D-glucopyranoside (18),syringin (19),elentheroside E (20),4-(1,2,3-trihydroxypropyl)-2,6-dimethoxyphenyl-I-O-β-D-glucopyranoside (21),and coniferin (22).ConclusionCompounds 1-6 are originated from Astragalus membranceus,compounds 7-18 are originated from Panax ginseng,and compounds 19-22 are originated from Acanthopanax senticosus by LC-MS analysis.Compound 1 is a new compound.

Simultaneous determination of five glycosides in Aidi injection by RP-HPLC-ELSD / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a RP-HPLC-ELSD method for the simultaneous determination of five glycosides in Aidi injection.</p><p><b>METHOD</b>The separation was carried out at 35 degrees C on a Kromasil C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water as the mobile phases in gradient mode. The flow rate was 1.0 mL x min(-1). The SHIMADZU ELSD-LT II detector was used. The detector temperature was at 40 degrees C and the pressure of carrier gas N2 was 0.35 MPa.</p><p><b>RESULT</b>The linear response (the natural logarithm of peak areas with corresponding mass) ranges were 21.2-212 mg x L(-1) for syringin, 30.8-308 mg x L(-1) for ginsenoside Rg1, 24.8-248 mg x L(-1) for ginsenoside Re, 25.6-256 mg x L(-1) for ginsenoside Rb1, 31.2-312 mg x L(-1), for astragaloside IV, respectively. The average recoveries (n = 9) of five glycosides were greater than 97.6%, and RSD were less than 1.1%.</p><p><b>CONCLUSION</b>The results demonstrated that this method had adequate accuracy and selectivity to measure the concentrations of five glycosides in Aidi injection.</p>

Chemical constituents of roots of Boehmeria nivea / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents from roots of Boehmeria nivea.</p><p><b>METHOD</b>The constituents were isolated by repeated column chromatography and preparative liquid chromatography; and their structures were elucidated by chemical properties and spectroscopic analyses.</p><p><b>RESULT</b>Seven compounds were isolated and their structures were identified as tormentic acid (1), hederagenin (2), maslinic acid (3), 2alpha-hydroxyursolic acid (4), trans-p-hydroxycinamic acid (5), 2,4,4'-trihydroxychalcone (6), rutin (7).</p><p><b>CONCLUSION</b>Compounds 1-6 were obtained from this genus for the first time.</p>

Simultaneous Determination of the Contents of Costunolide and Dehydrocostuslactone in Xuedan Weichang Pill by RP-HPLC / 中国药房

OBJECTIVE:To establish an RP-HPLC method for simultaneous determination of the contents of costunolide and dehydrocostuslactone in Xuedan weichang pill. METHODS:The determination was performed on Shim-pack VP-ODS column(150 mm?4.6 mm,5 ?m) with methanol-water(60∶40) as mobile phase and the detection wavelength was set at 225 nm.RESULTS:The linear ranges of costunolide and dehydrocostuslactone were 13.6～272.0 ?g?mL-1 and 11.25～225.0 ?g?mL-1,respectively,and their average recoveries were 99.47% and 99.62%,respectively with RSD of 1.75%(n=6) and 1.14%(n=6),respectively. CONCLUSION:The method established is simple,feasible,accurate and reliable,and it is applicable for the quality control of Xuedan weichang pill.

Preparation of osmotic pump-controlled release tablets of total flavones in Lysimachia clethroides / 中草药

Objective To prepare osmotic pump-controlled release tablets of total flavones in Lysimachia clethroides.Methods Two components of the extract from L.clethroides,rutin and naringenin-7-O-glucoside were used to evaluate the release behavior of osmotic pump controlled release tablets.Single factor investigation was carried out on the membrane compositions and orifice variables,and uniform design was used to optimize the formulation of coating mambrane.Results The membrane weight,PEG400 content,and dibutyl phthalote(DBP) content were the main factors influencing the drug release,and based on 45% and 8.5% of cellulose acetate,respectively,to prepare osmotic pump-controlled release tablets could achieve the desired zero-order release profile.Conclusion The formulation and technology are simple and easy to be carried out.Osmotic pump-controlled release tablets have a stable drug release bahavior and a good reproducibility.

Chemical constituents in roots of Boehmeria nivea / 中草药

Objective To investigate the chemical constituents in the roots of Boehmeria nivea.Methods The constituents were isolated by repeated column chromatography and their structures were elucidated by chemical properties and spectroscopic analyses.Results Seven compounds were isolated and their structures were identified to be daucosterol-10,13-eicosdienoate(Ⅰ),daucosterol(Ⅱ),?-sitosterol(Ⅲ),olein(Ⅳ),betulinic acid(Ⅴ),oleanolic acid(Ⅵ),19?-hydroxyursolic acid(Ⅶ).Conclusion Compound Ⅰ is a new compound named niveain A,compound Ⅳ is obtained from the plants of Boehmeria Jacq.for the first time.