Effects of Chinese herbs on multiple ion channels in isolated ventricular myocytes

Background Shensong Yangxin (SSYX) is one of the compound recipe of Chinese materia medica. This study was conducted to investigate the effects of SSYX on sodium current (I(Na)), L-type calcium current (I(Ca,L)), transient outward potassium current (I(to)), delayed rectifier current (I(K)), and inward rectifier potassium currents (I(K1)) in isolated ventricular myocytes. Methods Whole cell patch-clamp technique was used to study ion channel currents in enzymatically isolated guinea pig or rat ventricular myocytes. Results SSYX decreased peak I(Na) by (44.84±7.65)% from 27.21±5.35 to 14.88±2.75 pA/pF (n=5, P<0.05). The medicine significantly inhibited the I(Ca,L). At concentrations of 0.25, 0.50, and 1.00 g/100 ml, the peak I(Ca,L) was reduced by (19.22±1.10)%, (44.82±6.50)% and (50.69±5.64)%, respectively (n=5, all P<0.05). SSYX lifted the I-V curve of both I(Na) and I(Ca,L) without changing the threshold, peak and reversal potentials. At the concentration of 0.5%, the drug blocked the transient component of I(to) by 50.60% at membrane voltage of 60 mV and negatively shifted the inactive curve and delayed the recovery from channel inactivation. The tail current density of I(K) was decreased by (30.77±1.11)% (n=5, P<0.05) at membrane voltage of 50 mV after exposure to the medicine and the time-dependent activity of I(K) was also inhibited. Similar to the effect on I(K), the SSYX inhibited I(K1) by 33.10% at the test potential of –100 mV with little effect on reversal potential and the rectification property. Conclusions The experiments revealed that SSYX could block multiple ion channels such as I(Na) I(Ca,L), I(k), I(to) and I(K1), which may change the action potential duration and contribute to some of its antiarrhythmic effects.     

The electrophysiologic effects of endothelin. A patch clamp study in guinea pig ventricular myocytes.

To study the electrophysiologlc effects of endothelin-1 (ET-I), we used patch clamp and glass microelectrode techniques to investigate the effects of ET-1 on cardiac L-Ica, Ik and lk_2 in guinea pig ventricular myocytes. The prolongation of APD_50 was induced and EADs was triggered by 50 nM ET-I perfusion. L-Ica and Ik were enhanced by various ET-I concentration from I to 50 nM with dose-dependence. Their steady-state activations of L-Ica and Ik shifted left with ET-1 concentration increments. ET-1 elicited a kind of GTP- dependent inward rectifier K~ current having a mean conductance of 82.36± 1.27 pS. The open time and close time (both interburst intervals and burst durations ) abbreviated with ET-1 concentration increase. The results suggested that EADs-ET evoked was ascribed to the prolongation on the plateau level, which resulted from L-Ica inhancement. The ET- evoked inward rectifier K~ current should be further studied.     

Analysis of the Inhibitory Effect of Gypenoside on Na~ , K~ -ATPase in Rats' Heart and Brain and Its Kinetics

Objective:To study the effects of gypenoside(Gyp) on the activity of microsomal Na ,K -ATPase in rat's heart and brain in vitro.Methods:The microsomal Na ,K -ATPase was prepared from rat's heart and brain by differential centrifugation.The activity of microsomal Na ,K -ATPase was assayed by colorimetric technique.Enzyme kinetic analysis method was used to analyze the effect of Gyp on the microsomal Na ,K -ATPase of rats.Results:Gyp reversibly inhibited the brain and heart's microsomal Na ,K -ATPase in a concentration-dependent manner,and showed a more potent effect on enzyme in the brain.The IC50 of Gyp for the heart and brain were 58.79±8.05 mg/L and 52.07± 6.25 mg/L,respectively.The inhibition was enhanced by lowering the Na ,or K concentrations or increasing the ATP concentration.Enzyme kinetic studies indicated that the inhibitory effect of Gyp on the enzyme is like that of competitive antagonist of Na ,the counter-competitive inhibitor for the substrate ATP,and the mixed-type inhibitor for K .Conclusion:Gyp displays its cardiotonic and central inhibitory effects by way of inhibiting heart and brain's microsomal Na ,K -ATPase activities in rats.     

Cardioprotective effects of simvastatin on reversing electrical remodeling induced by myocardial ischemia-reperfusion in normocholesterolemic rabbits

Background Recent studies have revealed that pretreatment with statin is effective in preventing arrhythmia, but its electrophysiological mechanism is unclear. This study was conducted to investigate the cardioprotective effects of simvastatin on reversing electrical remodeling in left ventricular myocytes of rabbit heart undergoing ischemia-reperfusion, so as to explore the ionic mechanism responsible for the anti-arrhythmic effect of statin. Methods Forty-five rabbits were randomly divided into three groups： ischemic-reperfusion group （I-R）, simvastatin intervention group （Statin） and sham-operated control group （CON）. Anesthetized rabbits were subjected to 30-minute ischemia by ligation of the left anterior descending coronary artery and a 60-minute reperfusion after a 3-day administration of oral simvastatin of 5 mg-kg^-1.d^-1 in the Statin group or a placebo in the I-R group. Single ventricular myocytes were isolated enzymatically from the epicardial zone of the infracted region dedved from the hearts in the I-R and Statin group and the same anatomical region in the CON animals. The whole cell patch-clamp technique was used to record membrane ionic currents, including sodium current （IRa）, L-type calcium current （Ica-L） and transient outward potassium current （Ito）. Simultaneously, the level of serum cholesterol was examined. Results There was no significant difference in the serum cholesterol concentration among the three groups. The peak IRa current density （at -30 mV） was significantly decreased in I-R （（22.46±5.32） pA/pF, n=12） compared with CON （（42.78±5.48） pA/pF, n=16, P〈0.01） and Statin （（40.66±5.89） pA/pF, n=15, P〈0.01）, while the peak IRa current density in the Statin group was not different from CON （P〉0.05）. The peak ICa-L current density （at 0 mV） was significantly increased in I-R （（4.34±0.92） pA/pF, n=15） compared with CON （（3.13±1.22） pA/pF, n=13, P〈0.05） and Statin （（3.46±0.85） pNpF, n=16, P〈0     

Modulation of BmKAS-1 and BmK1-3-2 to sodium channel in rat dorsal root ganglion neurons

Objective To investigate what effects BmKAS-1 (a polypeptide purified from the Chinese sc orpion Buthus martensi Karsch ［BmK］ and named as BmK activator of skeletal -muscle ryanodine receptor) and its upstream mixture BmK1-3-2 have on Na(+) c hannels in dorsal root ganglion (DRG) small diameter neurons.Methods The whole-cell patch-clamp technique was used to investigate the effects of BmKAS-1 and BmK1-3-2 on Na(+) current in rat small diameter DRG neurons.Results About 50% peak Na(+) current was suppressed by 10 μg/ml of BmK1- 3-2. 1.62 μg/ml of BmKAS-1 also blocked 50% peak Na(+) cu rrent, and there was an obvious dose-dependent relationship.Conclusion Both BmK1-3-2 and BmKAS-1 have a blocking effect on Na(+) channels, and this may one of the mechanisms for the analgetic effect of BmK1-3-2 and BmKAS-1.     

Effect of Matrine on Human Ether à go-go Related Gene (HERG) Channels Expressed in Chinese Hamster Ovary Cells

Objective: To observe the effect of matrine on human ether à go-go related gene (HERG) potassium channels expressed in Chinese hamster ovary (CHO) cells and investigate whether HERG channel is a new target of the pharmacological effect of matrine on arrhythmia and tumor. Methods: HERG channel potassium current in CHO cell was recorded using whole-cell patch-clamp technique, and the influence of matrine on the current was explored. Results: Matrine inhibited HERG potassium current in a dose-dependent manner, and the 50% inhibitory concentration (IC50) was 411±23 μmol/L. Matrine had no significant effect on the activation kinetics, and mainly blocked HERG channels in their closed state. Conclusions: The blocking effect of matrine on HERG channels might be one of the mechanisms against arrythmias and tumors. Unlike most other blockers exerting blocking effect at the intracellular sites by entering the cell with the opening of HERG channel, matrine blocked HERG channels at the extracellular sites.     

Effects of K. Lysolecithin on Blood Levels of Monoamines in Mice

In this research, Lysolecithin-a substance made with 100% natural ingredients - was given to ICR mice as medication to measure its periodic effect on the noradrenalin (NA), dopamine (DA), and serotonin (5-HT) levels of the brain. Both ICR and SAM mice were separated into two groups - control group and Lysolecithin (K. Lysolecithin: hydrolytic lysolecithin) medicated group, and given 1-week preparation period. The K. Lysolecithin group was given 500mg/kg of K. Lysolecithin at 0.2mL per dosage for 4 weeks, and the control group was given the same amount of dosage of water during the same period. NA, DA and 5-HT concentrations were measured from the blood before medication and 8 weeks / 12 weeks / 16 weeks after the first medication. For the SAM mice, 8 weeks after they were medicated with K .Lysolecithin, Morris Water Maze Test was conducted for 7 consecutive days and then the concentrations were measured by drawing blood from the heart. The K. Lysolecithin medicated group showed a tendency to have a statistically significant higher concentrations of 5-HT and NA in the blood. Also, periodic examination showed that the monoamine levels were highest in the 12th week and declined thereafter.     

Contribution of Water and Lipid Soluble Substances in the Relaxant Effects of Tymus Vulgaris Extract on Guinea Pig Tracheal Smooth Muscle in Vitro

Objective:To examine the relaxant effects of hydro-ethanolic,macerated aqueous(MA) and lipidfree macerated aqueous(LFMA) extract of Tymus vulgaris on tracheal chains of guinea pigs.Methods:The relaxant effects of five cumulative concentrations of each extract(0.4,0.8,1.2,1.6 and 2.0 g/100 mL) were compared with saline as negative control and five cumulative concentrations of theophylline(0.1,0.2,0.4,0.6 and 0.8 mmol/L) on precontracted tracheal smooth muscle of guinea pig with 60 mmol/L KCI(group 1) and 10 μmol/L methacholine(group 2,n=6 for each group).Results:In group 1 all concentrations of theophylline,three higher concentrations of hydro-ethanolic,two concentrations of LFMA and last concentration of MA extracts showed significant relaxant effects compared with that of saline(P0.05 or P0.01).Two lower concentrations of LFMA and all concentrations of MA except higher one caused contraction compared with saline(P0.05 or 0.01).In group 2 experiments,all concentrations of theophylline,hydro-ethanolic,MA and LFMA extracts showed significant relaxant effects compared to that of saline(P0.05 or P0.01).In both groups,the relaxant effect of all concentrations of hydro-ethanolic extract were significantly higher than most concentrations of others(P0.05 or P0.01).The relaxant effect of different concentrations of three extracts were significantly greater in group 2 compared with group 1 experiments(all P0.01).There were significantly positive correlations between the relaxant effects and concentrations for theophylline and ail extracts in both groups(P0.05 or P0.01).Conclusion:Hydro-ethanolic extract has a potent weaker relaxant effect for other extracts from Tymus vulgaris on tracheal chains of guinea pigs.     

Effect of Chengzai Pill on the L-Type Ca^2＋ Channel Currents of Osteoblasts Pretreated with Methylprednisolone

Objective: To observe the effect of the serum containing Chengzai Pill on the L-type voltage-sensitive calcium channels current (L-VSCCsC) of osteoblastic MC3T3-E1 cells pretreated with methylprednisolone (mPSL). Methods: A control group, a model group, a low dose group and a high dose group were set up. The whole cell patch clamp technique was used to record L-VSCCsC of 10 osteoblastic MC3T3-E1 cells in each group and their peak currents were determined. Results: The peak current of the control group was 0.2284±0.0209 nA; the peak current of the model group was 0.1839±0.0179 nA; decreased by 19.5% as compared with the control group (P<0.01); the peak current of the low and high dose groups was 0.2526± 0.0093 nA and 0.2671±0.0120 nA respectively, increased by 37.4% and 45.2% as compared with the model group (P<0.01); the difference between the low and high dose groups was P<0.05. Conclusion: 1. mPSL inhibits L-VSCCsC of osteoblasts; and 2. The serum containing Chengzai Pill increases L-VSCCsC of osteoblasts pretreated with mPSL.     

Antiviral Effect of Chinonin against Herpes Simplex Virus

In order to investigate the antiviral effect of chinonin against Herpes simplex virus (HSV), the encephalitis model in mice and skin infection model in guinea pigs were established by HSV-Ⅰ and HSV-Ⅱ infection respectively. Acyclovir was used as the positive reference drug to evaluate the antiviral capacity of chinonin. Chinonin showed an obvious therapeutic effect on encephalitis in mice at doses of 25 and 50 mg/kg. At both dosages, chinonin demonstrated stronger protection than acyclovir (1 and 5 mg/kg) to the infected mice from death. It was also found that chinonin could treat the skin infection in guinea pigs effectively. The therapeutic effect of chinonin was similar to that of acyclovir (5 mg/kg) at 25 mg/kg but obviously better than that at 50 and 75 mg/kg. In conclusion, chinonin is a potential candidate for the treatment against HSV.     

Effects of gastrointestinal peptides on formation of gallstone in guinea pigs.

In light of the effects of gastrointestinal (GI) peptides on bile secretion and biliary tract mobility, we studied the effects of GI peptides on gallstone formation in guinea pigs fed on low protein lithogenic diet. The peptides under study included cholecystokinin octapeptide (CCK-8), vasoactive intestinal peptide (VIP), somatostatin (SRIF), secretin (SEC), and neurotensin (NT). Hepatic bile flow, electrolytes, and other bile components were also measured. It was found that CCK-8 and VIP suppressed the formation of gallstones and increased hepatic bile flow and Na+, K+, Cl- output significantly. On the other hand, SRIF significantly promoted gallstone formation. The rates of gallstone formation in CCK-8, VIP, and SRIF treated guinea pigs were 15.4%, 23.5%, and 88.0%, respectively, in contrast to 56.8% in the control group. The inhibitory effect of CCK-8 and promoting effect of SRIF on gallstone formation were dose-dependent.

     

ANTAGONISTIC EFFECT OF RADIX PUERARIAE ON THE OTOTOXICITY OF SODIUM ETHOCRYNATE ANEXPERIMENTAL STUDY

Comparision was made on the amplitude of coch-lear microphonics(CM)and the threshold of activepotential(APN_1)of guinea pigs before and after in-travenous injection of sodium ethocrynate,to observethe antagonistic effect of Radix Puerariae on theototoxicity of ethoerynie acid.The results of our ex-periments confirmed that daily intragastric admini-stration of Radix Puerariae extract(4 ml/kg)for 30days produced a marked antagonistic effect.     

Reversal of Multidrug Resistance and Inhibition of Phosphorylation of AKT in Human Ovarian Cancer Cell Line by Wild-type PTEN Gene

The reversing effect of wild-type PTEN gene on resistance of C13K cells to cisplatin and its inhibitory effect on the phosphorylation of protein kinase B (AKT) were studied. The expression of PTEN mRNA and protein in OV2008 cells and C13K cells were semi-quantitatively detected by using RT-PCR and Western blotting. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, Akt, p-Akt protein were ana- lyzed by Western blotting in PTEN-transfected and non-transfected C13K cells. Proliferation and chemosensitivity of cells to DDP were measured by MTT, and cell apoptosis was detected by flow cytometry after treatment with cisplatin. The expression of PTEN mRNA and protein in OV2008 cells were significantly higher than those in C13K cells. After transfection with PTEN gene for 48 h, the expression of PTEN mRNA and protein in C13K cells were 2.04±0.10, 0.94±0.04 respectively and the expression of p-Akt protein ( 0.94±0.07) was lower than those in control groups (1.68±0.14, 1.66±0.10) (P< 0.05). The IC50 of DDP to C13K cells transfected with PTEN (7.2±0.3 μmol/L) was obviously lower than those of empty-vector transfected cells and non-transfected cells (12.7±0.4 μmol/l, 13.0±0.3 μmol/L) (P<0.05). The apopototis ratio of wild-type PTEN-transfected, empty vector transfected and non-transfected C13K cells were (41.65±0.87)%, (18.61±0.70)% and (15.28 ±0.80)% respectively, and the difference was statistically significant (P<0.05). PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the ex- pression of PTEN and restore drug sensitivity to cisplatin in human ovarian cancer cell line C13K with multidrug-resistance by decreasing the expression of p-Akt.     

Frequency and Voltage-dependent Inhibition of Delayed Outward Potassium Current by Flecainide in Isolated Atrial Cell of Guinea Pig Heart

Effects of flecanide on membrane currents were studied using an isolated single atrial cell from guinea pig hearts. The tightseal cell clamp technique was used. In the current clamp condition, flecainide prol(?)nged significantly the atrial action potential (APD) with frequency dependence.Delayed outward K current (I_k) and outward tail current were specifically inhibited by flecainide in a frequency and concentration fashion. Flecainide inhibit(?)d I_k more strongly as the membrane potential become more positive from+10mV to+60mV.The value of I_k was attenuated to 0.973 nA from 1.105 nA of control and the value of tail current was reduced to 0.113 nA from 0.288 nA of control at 60 mV. The drug did not affect on the holding current.The effects of flecainide on the action potential and transmembrane ionic currents strongly suggest that the main mechanism of action of this agent is to inhibit Voltage-dependent potassium current. In the Voltage clamp condition, flecainide affected neither the conventional L type Ca~(2+) current nor the I~(k1) current significantly. Our research proved that F1e was not completely consistent with class Ic agents, because F1e could markedly increase the APD in the experiment.     

Effect of DRB on the Biological Characteristics of Human Laryngeal Carcinoma Hep-2 Cell Line

In order to study the effect of 5, 6-Dichloro-1-β-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68±0.19)%, (1.95±0.12)%, (8.51±0.26)%, (11.26±0.17)% and (14.99±0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concen-tration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.     

Inhibitory Effect of TanshinoneⅡA on TGF-β1-induced Cardiac Fibrosis

This study examined the effect of tanshinoneⅡA (TSNⅡA) on the cardiac fibrosis induced by transforming growth factor β1 (TGF-β1) and the possible mechanisms. Cardiac fibroblasts were isolated from cardiac tissues of neonatal Sprague-Dawley (SD) rats by the trypsin digestion and differential adhesion method. The cells were treated with 5 ng/mL TGF-β1 alone or pretreated with TSNⅡA at different concentrations (10–5 mol/L, 10–4 mol/L). Immunocytochemistry was used for cell identification, RT-PCR for detection of the mRNA expression of connective tissue growth factor (CTGF) and collagen type Ⅰ (COLⅠ), Western blotting for detection of the protein expression of Smad7 and Smad3, and immunohistochemistry and immunofluorescence staining for detection of the protein expression of phosphorylated Smad3 (p-Smad3), CTGF and COLⅠ. The results showed that TGF-β1 induced the expression of CTGF, COLⅠ, p-Smad3 and Smad7 in a time-dependent manner. The mRNA expression of CTGF and COLⅠ was significantly increased 24 h after TGF-β1 stimulation (P<0.01 for all). The protein expression of p-Smad3 and Smad7 reached a peak 1 h after TGF-β1 stimulation, much higher than the baseline level (P<0.01 for all). Pretreatment with high concentration of TSNⅡA resulted in a decrease in the expression of p-Smad3, CTGF and COLⅠ (P<0.01). The protein expression of Smad7 was substantially upregulated after pretreatment with two concentrations of TSNⅡA as compared with that at 2h post TGF-β1 stimulation (P<0.05 for low concentration of TSNⅡA; P<0.01 for high concentration of TSNⅡA). It was concluded that TSNⅡA may exert an inhibitory effect on cardiac fibrosis by upregulating the expression of Smad7, suppressing the TGF-β1-induced phosphorylation of Smad3 and partially blocking the TGF-β1-Smads signaling pathway.     

Injury of Mouse Brain Mitochondria Induced by Cigarette Smoke Extract and Effect of Vitamin C on It in vitro

Objective To investigate the toxicity of cigarette smoke extract (CSE) and nicotine on mouse brain mitochondria as well as the protective effect of vitamin C in vitro. Method Mouse brain mitochondria in vitro was incubated with CSE or nicotine in the absence or presence of vitamin C for 60 minutes, and the changes of mitochondrial function and structure were measured. Results CSE inhibited mitochondrial ATPase and cytochrome C oxidase activities in a dose-dependent manner.However, no significant changes in the peroxidation indices were observed when mitochondrial respiratory enzymes activity was inhibited, and protection of mitochondria from CSE-induced injury by vitamin C was not displayed in vitro. The effect of CSE on mouse brain mitochondria swelling response to calcium stimulation was dependent on calcium concentrations. CSE inhibited swelling of mitochondria at 6.5 μmol/L Ca^2 , but promoted swelling response at 250μmol/L Ca^2 . Nicotine, the major component of cigarette smoke, showed no significant damage in mouse brain mitochondria in vitro. The CSE treatment induced mitochondrial inner membrane damage and vacuolization of the matrix, whereas the outer mitochondrial membrane appeared to be preserved. Conclusion The toxic effect of CSE on brain mitochondria may be due to its direct action on enzymatic activity rather than through oxygen free radical injury. Nicotine is not the responsible component for the toxicity of CSE to brain mitochondria.     

The Apoptosis of Bovine Lens Epithelial Cells Induced by Proteasome Inhibitor MG132

To investigate the effect of proteasome inhibitor MG132 on the apoptosis of bovine lens epithelial cells （BLECs）, the cells were treated with MG132 at different concentrations for12, 24 and 36 h. The cell viability was analyzed by MTT assay and the effect of MG132 on the apoptosis of BLECs was assessed by flow cytometry （FCM）. The results showed that after treatment for the same period, the inhibitory effect of MG132 on BLECs proliferation was enhanced with the increment of the concentration of MG132 （0, 2, 5, 10, μmol/L） （P〈0.05）. The 50% inhibiting concentration （IC50） was 2.03 μmol/L when the BLECs were treated with MG132 for 36 h. MG132 also induced the apop- tosis of BLECs obviously. FCM showed that the apoptosis index of the cells treated by MG132 at 2 μmol/L for 12 h was （20.24±1.51）%, and that of the control was （0.98±0.20）% respectively （P〈0.01, n=3）. It was concluded that MG132 could lead to apoptosis of BLECs. The decrease of proteasome activity may play an important role in the formation and development of cataract.     

Effects of ranitidine on contraction and action potentials in normal and potassium depolarized isolated papillary muscles of the guinea pig

The purpose of these experiments was to investigate the possible ionicbases for ranitidine (Ran) in isolated papillary muscles of the guinea pig.Ran,when the concentration was below 300μmol/L,had no influence on contraction ofnormal muscles and only above 0.3 mmol/L,it showed weak negative inotropic ef-fect.The contractile activity elicited by histamine in potassium depolarized mus-cles was depressed by Ran markedly and dose-dependently.The IC_(50) of Ran was0.273μmol/L.This depressant effect was reversed by an elevation of externalCa~(2+)concentration to 7.7mmol/L.The contraction induced by isoproterenol inpotassium depolarized preparations was not altered by Ran.Ran at theconcentrations of 1,10 and 100μmol/L failed to show influences on fast actionpotentials.However,Ran at 0.5μmol/L time-dependently inhibited slow action po-tentials.It is concluded that the ionic mechanism of Ran might be attributed toantagonized H_2 receptor of myocardium which in turn decreases Ca~(2+) inward cur-rent.     

The anti-HSV-2 effect of alumen: In vitro and in vivo experimental studies

This study investigated the anti-HSV-2 effect of alumen through in vitro and in vivo expe-riments.Viable cell counting was employed to assess the toxicity of alumen on Vero cells.The inhibition rate of HSV-2 was defined as the cytopathic effect(CPE)of the cells infected with the virus.Alumen suppositories of different concentrations were vaginally applied to the guinea pigs which were then infected with HSV-2 via a vaginal route.The clinical symptoms were observed and the local virus titer calculated.The results showed that alumen had an in vitro anti-HSV-2 effect by means of antiviral dup-lication,direct killing of the virus,and antiviral adsorption.Alumen suppositories of different concen-trations could reduce or completely inhibit HSV-2 infection in guinea pigs.It was concluded that alumen had an in vitro anti-HSV-2 effect through multiple approaches and it could suppress in vivo vaginal HSV-2 infection of guinea pig to some extent.