Method of treating a disease associated with potassium ion channel

Abstract

An application of a combination of traditional Chinese medicine in the preparation of a potassium ion channel modulating agent. The combination of traditional Chinese medicine is made up of traditional Chinese medicine raw materials according to the following weight proportion: Coptidis Rhizoma 250˜450; Pinelliae Rhizoma 150˜350; Poria 150˜450; Aurantii Fructus Immaturus 62˜265; Dichroae Radix 150˜350; Nelumbinis Plumula 10˜70; Sophorae Flavescentis Radix 150˜350; Artemisiae Annuae Herba 150˜350; Ginseng Radix Et Rhizoma 65˜265; Ophiopogonis Radix 150˜350; and Glycyrrhizae Radix Et Rhizoma 62˜265.

Claims

1. A method of treating a disease associated with potassium ion channel in a subject, comprising: preparing a traditional Chinese medicine composition, wherein the concentration of the traditional Chinese medicine composition is ≥2 mg/ml and the traditional Chinese medicine composition is made up of traditional Chinese medicine raw material according to the following weight proportion: TABLE-US-00006 Coptidis Rhizoma 250~450 Sophorae Flavescentis 150~350 Radix Pinelliae Rhizoma 150~350 Artemisiae Armuae Herba 150~350 Poria 150~450 Ginseng Radix Et Rhizoma  65~265 Aurantii Fructus  65~265 Ophiopogonis Radix 150~350 Immaturus Dichroae Radix 150~350 Glycyrrhizae Radix Et   65~265, Nelumbinis Plumula 10~70 Rhizoma wherein preparing the traditional Chinese medicine composition comprises: (1) Picking The foreign matters and non medicinal parts out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizome, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizome, Pinelliae Rhizoma, Ophiopogonis Radix, and Nelumbinis Plumula, respectively, and then setting aside until use; (2) Rinsing the residue from step (1) with flowing water and cutting into slices or segments; moistening Ophiopogonis Radix with a flowing water then crushing to be flat and setting aside until use; (3) Drying The crude drugs of step (2) at 70˜80° C. and setting aside until use; (4) Extracting Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Nelumbinis Plumula and Sophorae Flavescentis Radix by reflux with 60% ethanol for two times, adding eight times amount of solvent for the first time and six times for the second time, 1.5 h for each time, combining the extracts, filtrating, and concentrating the filtrate in vacuum at 0.04 Kpa and recovering ethanol at 70° C., continuing to condense to obtain thick paste with the relative density of 1.38 when measured at 60° C., then drying into dry paste at 80° C. and setting aside until use; (5) Extracting Ginseng Radix Et Rhizoma, Pinelliae Rhizoma and Poria by reflux with 70% ethanol for two times, adding eight times amount of solvent for the first time and six times for the second time, 2 h for each time, combining extracts, filtrating, and collecting the filtrate and setting aside until use; (6) Combining The residue from step 5 with Ophiopogonis Radix, Artemisiae Annuae Herba and Glycyrrhizae Radix Et Rhizoma, then decocting with water for two times, adding ten times amount of water for the first time and eight times for the second time, 1 h for each time, combining decoction, filtrating, concentrating the filtrate to obtain liquid extract with the relative density ranging from 1.05 to 1.06 when measured at 80° C. and then adding 95% ethanol until the alcohol content reaches 70%, stirring well and standing for 24 h, removing and filtering the supernatant, collecting the filtrate and setting aside until use; (7) Merging the filtrates of steps 5 and 6, concentrating the filtrate in vacuum at 0.04 Kpa and recovered ethanol at 70° C., continuing to condense to thick paste with the relative density of 1.38 when measured at 60° C., then drying to dry extract at 80° C. and setting aside until use; and (8) Combining the dry extract of steps 4 and 7 and crushing into 100 mesh fine powder, adding a proper amount of dextrin, then mixing, packing into capsules or prepared preparation with medicinal excipients; and administering an effective amount of the prepared traditional Chinese medicine composition to the subject.

2. The method of claim 1, wherein the disease is arrhythmia.

3. The method of claim 1, wherein the disease is ventricular, atrial or ventricular arrhythmia.

4. The method of claim 1, wherein the traditional Chinese medicine composition prolongs the action potential duration, regulates potassium channel and inhibits various potassium currents.

5. The method of claim 1, wherein the traditional Chinese medicine composition is a tablet, capsule, granule, oral liquid or pill.

6. A method of treating a disease associated with potassium ion channel in a subject comprising: preparing a traditional Chinese medicine composition, wherein the concentration of the traditional Chinese medicine composition is ≥2 mg/ml and the traditional Chinese medicine composition is made up of traditional Chinese medicine raw material according to the following weight proportion: TABLE-US-00007 Coptidis Rhizoma 334 g Sophorae Flavescentis Radix 250 g Pinelliae Rhizoma 250 g Artemisiae Annuae Herba 250 g Poria 250 g Ginseng Radix Et Rhizoma 167 g Aurantii Fructus 167 g Ophiopogonis Radix 250 g Immaturus Dichroae Radix 250 g Glycyrrhizae Radix Et Rhizoma  167 g, Nelumbinis Plumula  42 g wherein preparing the traditional Chinese medicine composition comprises: (1) Picking The foreign matters and non medicinal parts out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, Ophiopogonis Radix, and Nelumbinis Plumula, respectively, and then setting aside until use; (2) Rinsing the residue from step (1) with flowing water and cutting into slices or segments; moistening Ophiopogonis Radix with a flowing water then crushing to be flat and setting aside until use; (3) Drying The crude drugs of step (2) at 70˜80° C. and setting aside until use; (4) Extracting Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Nelumbinis Plumula and Sophorae Flavescentis Radix by reflux with 60% ethanol for two times, adding eight times amount of solvent for the first time and six times for the second time, 1.5 h for each time, combining the extracts, filtrating, and concentrating the filtrate in vacuum at 0.04 Kpa and recovering ethanol at 70° C., continuing to condense to obtain thick paste with the relative density of 1.38 when measured at 60° C., then drying into dry paste at 80° C. and setting aside until use; (5) Extracting Ginseng Radix Et Rhizoma, Pinelliae Rhizoma and Poria by reflux with 70% ethanol for two times, adding eight times amount of solvent for the first time and six times for the second time, 2 h for each time, combining extracts, filtrating, and collecting the filtrate and setting aside until use; (6) Combining The residue from step 5 with Ophiopogonis Radix, Artemisiae Annuae Herba and Glycyrrhizae Radix Et Rhizoma, then decocting with water for two times, adding ten times amount of water for the first time and eight times for the second time, 1 h for each time, combining decoction, filtration, concentrating the filtrate to obtain liquid extract with the relative density ranging from 1.05 to 1.06 measured at 80° C. and then adding 95% ethanol until the alcohol content reaches 70%, stirring well and standing for 24 h, removing and filtering the supernatant, collecting the filtrate and setting aside until use; (7) Merging the filtrates of steps 5 and 6, concentrating the filtrate in vacuum at 0.04 Kpa and recovering ethanol at 70° C., continuing to condense to thick paste with the relative density of 1.38 when measured at 60° C., then drying to dry extract at 80° C. and setting aside until use; and (8) combining the dry extract of steps 4 and 7 and crushing into 100 mesh fine powder, adding a proper amount of dextrin, then mixing, packing into capsules or prepared preparation with medicinal excipients; and administering an effective amount of the prepared traditional Chinese medicine composition to the subject.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1. The prolongation effects of the combination of traditional Chinese medicine at 2 mg/ml on the action potential.

(2) The abscissa indicates the duration of medication (Unit: min), the ordinate indicates the decay time of the action potential (Unit: msec). 1 indicates the duration of medication at 2 mg/ml is from 0.65 min to 10.16 min.

(3) FIG. 2. The action potential diagram of the combination of traditional Chinese medicine at 2 mg/ml and 4 mg/ml.

(4) The abscissa indicates the duration of medication (Unit: msec), the ordinate represents the action potential (Unit: mV). Line 1 indicates the relationship between the action potential and the time of the control group, and line 2 represents the relationship between the action potential and the time of the drug group (2 mg/ml), and line 3 indicates the relationship between the action potential and the time after washout of the cells.

(5) FIG. 3. The attenuation relationship of the action potential of the combination of traditional Chinese medicine at different concentrations (1 mg/ml, 2 mg/ml, and 4 mg/ml).

(6) The abscissa indicates different concentration group, the ordinate indicates the action potential decrement. Column 1 is the attenuation of action potential of the control group, the attenuation of action potential is set to 1 in the control group. Column 2 is the attenuation of action potential of the drug group (1 mg/ml). Column 3 is the attenuation of action potential of the drug group (2 mg/ml). Column 4 is the attenuation of action potential of the drug group (4 mg/ml).

DETAIL DESCRIPTION OF PREFERRED EMBODIMENT

(7) In order to facilitate the application of the combination of traditional Chinese medicine of the present invention to the electrophysiological study of cardiac myocytes, the combination of traditional Chinese medicine is prepared into freeze-dried powder or extract according to the proportion of prescriptions.

(8) The traditional Chinese medicine raw materials used in the embodiment are commercially available.

Embodiment 1

(9) [Prescription]

(10) TABLE-US-00003 Coptidis Rhizoma 334 g Sophorae Flavescentis Radix 250 g Pinelliae Rhizoma 250 g Artemisiae Annuae Herba 250 g Poria 250 g Ginseng Radix Et Rhizoma 167 g Aurantii Fructus 167 g Ophiopogonis Radix 250 g Immaturus Dichroae Radix 250 g Glycyrrhizae Radix Et Rhizoma 167 g Nelumbinis Plumula  42 g

(11) 1). The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, Ophiopogonis Radix, and Nelumbinis Plumula, respectively, and then set aside until use;

(12) The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, respectively, and then rinsed (sprayed) or moistened (slightly moistened) with flowing water and cut into slices or segments; Ophiopogonis Radix is moistened with a flowing water then crashed as flate and set aside until use. Dried at 70˜80° C.

(13) 2). Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Nelumbinis Plumula and Sophorae Flavescentis Radix are washed with 20 kg tap water for two times, and rinsed with 10 kg purified water for two times, then extracted by reflux with 60% ethanol for two times, add 8.4 L for the first time and 6.3 L for the second time, 1.5 h for each time. Combined extract, filtrated, and the filtrate is concentrated in vacuum at 0.04 Kpa and recovered ethanol at 70° C. Then 6.3 L injection water is added, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(14) 3). Group B, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma and Poria are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, then extracted by reflux with 70% ethanol for two times, add 5.3 L for the first time and 4 L for the second time, 2 h for each time. Combined extract, filtrated, the filtrate and residue are set aside until use, respectively;

(15) 4). Group C, Ophiopogonis Radix, Artemisiae Annuae Herba and Glycyrrhizae Radix Et Rhizoma are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, then combined with the residue in group B, decocted with injection water for two times, add 6.7 L for the first time and 5.3 L for the second time, 1 h for each time. Combined the filtrates for the second time and discarded the residue, the filtrate is concentrated to obtain liquid extract with the relative density ranging from 1.05 to 1.06 (80° C.) and then 95% ethanol is added to the alcohol content reaching 70%, stirred well and stand for 24 h. The supernatant is combined with the filtrate in group B, and recovered ethanol at 70° C. Then add 4 L injection water, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(16) 5). Combined the supernatant of group A and C, then concentrated to obtain liquid extract with the relative density of 1.10 (60° C.) by rotating film evaporator and set aside until use;

(17) 6). Fixed the liquid extract with the relative density of 1.10 (60° C.) in pallet, freeze-dried for 20 h below −40° C. with the vacuum below 15 pa and the heating temperature from −20° C. to 80° C. Taken out and collected the freeze-dried solids, crushed, mixed, then 590 g lyophilized powder is obtained.

Embodiment 2

(18) [Prescription]

(19) TABLE-US-00004 Coptidis Rhizoma 334 g Sophorae Flavescentis Radix 250 g Pinelliae Rhizoma 250 g Artemisiae Annuae Herba 250 g Poria 250 g Ginseng Radix Et Rhizoma 167 g Aurantii Fructus 167 g Ophiopogonis Radix 250 g Immaturus Dichroae Radix 250 g Glycyrrhizae Radix Et Rhizoma 167 g Nelumbinis Plumula  42 g

(20) 1). The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, Ophiopogonis Radix, and Nelumbinis Plumula, respectively, and then set aside until use;

(21) The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, respectively, and then rinsed (sprayed) or moistened (slightly moistened) with flowing water and cut into slices or segments; Ophiopogonis Radix is moistened with a flowing water then crashed as flate and set aside until use. Dried at 70˜80° C.

(22) 2). Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Nelumbinis Plumula and Sophorae Flavescentis Radix are washed with 20 kg tap water for two times, and rinsed with 10 kg purified water for two times, then extracted by reflux with 60% ethanol for two times, add 8.4 L for the first time and 6.3 L for the second time, 1.5 h for each time. Combined extract, filtrated, and the filtrate is concentrated in vacuum at 0.04 Kpa and recovered ethanol at 70° C. Then 6.3 L injection water is added, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(23) 3). Group B, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma and Poria are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, and then extracted by reflux with 70% ethanol for two times, add 5.3 L for the first time and 4 L for the second time, 2 h for each time. Combined extract, filtrated, the filtrate and residue are set aside until use, respectively;

(24) 4). Group C, Ophiopogonis Radix, Artemisiae Annuae Herba and Glycyrrhizae Radix Et Rhizoma are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, then combined with the residue in group B, decocted with injection water for two times, add 6.7 L for the first time and 5.3 L for the second time, 1 h for each time. Combined the filtrates for the second time and discarded the residue, the filtrate is concentrated to obtain liquid extract with the relative density ranging from 1.05 to 1.06 (80° C.) and then 95% ethanol is added to the alcohol content reaching 70%, stirred well and stand for 24 h. The supernatant is combined with the filtrate in group B, and recovered ethanol at 70° C. Then add 4 L injection water, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(25) 5). Combined the supernatant of group A and C, then concentrated to obtain liquid extract with the relative density of 1.10 (60° C.) by rotating film evaporator and set aside until use;

(26) 6). Heated the liquid extract with the relative density of 1.10 (60° C.) in water bath into an extract without mobility, mixed, then it is obtained.

Embodiment 3

(27) [Prescription]

(28) TABLE-US-00005 Coptidis Rhizoma 334 g Sophorae Flavescentis Radix 250 g Pinelliae Rhizoma 250 g Artemisiae Annuae Herba 250 g Poria 250 g Ginseng Radix Et Rhizoma 167 g Aurantii Fructus 167 g Ophiopogonis Radix 250 g Immaturus Dichroae Radix 250 g Glycyrrhizae Radix Et Rhizoma 167 g Nelumbinis Plumula  42 g

(29) 1). The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, Ophiopogonis Radix, and Nelumbinis Plumula, respectively, and then set aside until use;

(30) The foreign matters and non medicinal parts are picked out from Artemisiae Annuae Herba, Sophorae Flavescentis Radix, Glycyrrhizae Radix Et Rhizoma, Poria, Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma, respectively, and then rinsed (sprayed) or moistened (slightly moistened) with flowing water and cut into slices or segments; Ophiopogonis Radix is moistened with a flowing water then crashed as flate and set aside until use. Dried at 70˜80° C.

(31) 2). Coptidis Rhizoma, Aurantii Fructus Immaturus, Dichroae Radix, Nelumbinis Plumula and Sophorae Flavescentis Radix are washed with 20 kg tap water for two times, and rinsed with 10 kg purified water for two times, then extracted by reflux with 60% ethanol for two times, add 8.4 L for the first time and 6.3 L for the second time, 1.5 h for each time. Combined extract, filtrated, and the filtrate is concentrated in vacuum at 0.04 Kpa and recovered ethanol at 70° C. Then 6.3 L injection water is added, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(32) 3). Group B, Ginseng Radix Et Rhizoma, Pinelliae Rhizoma and Poria are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, then extracted by reflux with 70% ethanol for two times, add 5.3 L for the first time and 4 L for the second time, 2 h for each time. Combined extract, filtrated, the filtrate and residue are set aside until use, respectively;

(33) 4). Group C, Ophiopogonis Radix, Artemisiae Annuae Herba and Glycyrrhizae Radix Et Rhizoma are washed with 13 kg tap water for two times, and rinsed with 6.5 kg purified water for two times, then combined with the residue in group B, decocted with injection water for two times, add 6.7 L for the first time and 5.3 L for the second time, 1 h for each time. Combined the filtrates for the second time and discarded the residue, the filtrate is concentrated to obtain liquid extract with the relative density ranging from 1.05 to 1.06 (80° C.) and then 95% ethanol is added to the alcohol content reaching 70%, stirred well and stand for 24 h. The supernatant is combined with the filtrate in group B, and recovered ethanol at 70° C. Then add 4 L injection water, heated at 50° C.˜60° C., stirred to make it fully dissolved, filtrated while hot, stand for overnight, the supernatant is set aside until use;

(34) 5). Combined the supernatant of group A and C, then concentrated to obtain liquid extract with the relative density of 1.10 (60° C.) by rotating film evaporator and set aside until use;

(35) 6). Heated the liquid extract with the relative density of 1.10 (60° C.) in water bath into an extract without mobility. Dried into dry extract at 80° C., crushed into 100 mesh powder, 200 g dextrin is added, mixed, then it is obtained by putting into capsules.

Embodiment 4

(36) For the combination of traditional Chinese medicine in the present invention (lyophilized powder is obtained in embodiment 1), the electrophysiological test of myocardial cells is carried out.

(37) 1. Experimental Materials:

(38) (1) Experimental Animals:

(39) Male rats, weighting 250˜500 g, was provided by the experimental animal center of University of Oxford.

(40) (2) Drugs and Reagents

(41) The test powder of the present invention is obtained in embodiment 1.

(42) Precision weighing the test powder of the present invention and dissolved in the extracellular solution, the final concentration is 1 mg/ml, 2 mg/ml, 4 mg/ml, respectively. Reagents are purchased from Sigma Aldrich Co., Ltd (St. Louis, Mo., USA).

(43) (1) Solution:

(44) 1) Extracellular Solution (mmol/L):

(45) NaCl 112.0 g, KCl 10 g, KH.sub.2PO.sub.4 1.2 g, HEPES 10 g, MgSO.sub.4 5 g, NaHCO.sub.3 15 g, Taurine 30 g, Glucose 20 g, NaHCO.sub.3 24.0 g (pH 7.4). Continuously pass into 5% CO.sub.2 and 95% O.sub.2.

(46) 2) Electrode solution (mmol/L): KCl 140 g, MgCl.sub.2 1 g, EGTA 5.0 g, Hepes 10.0 g, Na.sub.2ATP 2.0 g (pH 7.2).

(47) 2. Research Technique: Whole Cell Voltage Clamp Technique is Used to Record the Action Potential of Single Ventricular Myocyte, and to Make the Testing Drug Solution Act on Cells or Wash Out Cells by Continuously Infused.

(48) 1) Isolation of Single Ventricular Myocyte from Rats

(49) Single ventricular myocyte is isolated from the heart of adult rats by enzyme digestion. The rats are stunned by the head crash, fixed on supine position after carotid artery bloodletting, and open the chest quickly and take out the heart. The heart is attached to the Waldorf perfusion system and the enzyme liquid is infused through the heart, and placed in calcium free solution at 36° C., continuously infused about 5 min. The constituents of calcium free solution are: NaCl 120 g, KCl 5.4 g, MgSO.sub.4 5 g, Pyruvic acid 5 g, Glucose 20 g, Taurine 20 g, HEPES 10 g (pH 6.96). The perfusate is added with 4 U/ml protease (Sigma type, XXIV) to replace calcium free perfusion solution. Infused for 2 min, the perfusate is switched again to the solution contained collagenase (Worthington 2, 0.3 mg/ml) and hyaluronidase (Sigma, 0.6 mg/ml). After 5 to 10 min, removed the heart from the cannula, discarded the atrium and cut the ventricles, put in a high potassium solution, the constituents are: (KOH 5 g, KCl 30 g, KH.sub.2PO.sub.4 30 g, MgSO.sub.4 3 g, Glutamate 50 g, Taurine 20 g, EGTA 0.5 g, HEPES 10 g, Glucose 10 g (pH 7.4). The tissue should be filtered through four layers of gauze, the filtrate is centrifuged at 1500 rpm for 2 min Precipitated cells, discarded supernatant, potassium solution is added and centrifuged at 1500 rpm for 2 min, the cells are placed in a high potassium solution, kept at 4° C. until use.

(50) 2) Electrophysiological Techniques and Statistics

(51) Put the separated cells into the cell bath, continuously infused (22˜24° C.), the cell membrane potential is recorded by whole cell voltage clamp technique, and an 200B axonal patch clamp electrode amplifier with borosilicate glass electrode is used. The resistance is 1.5˜3Ω, when the conductive liquid is added to the electrode. The current is recorded by 9th PCLAMPEX software (American Axon Corporation), PCLAMPFIt software (American Axon Corporation) for data analysis, and draw with Origin software (American, OriginLab Corporation). The drug is dissolved in the perfusate, administered after 3 min of infusion. The data are expressed as the mean SD, the effect of the drug is the difference between the control and administered groups. To compare the two groups, Student's t-test is employed.

(52) 3. Research Results:

(53) (1) The results show that: the combination of traditional Chinese medicine obviously prolongs the action potential interval of the myocardial cells, and shows the function of regulating potassium channel: 45.4 ms±4.9 for control, 52.2 ms±4.5 (P<0.01, n=7) for the combination of traditional Chinese medicine (2 mg/ml). After administration of the drug, the action potential returns to the state before administration. As shown in FIG. 1, the results show that the drug obviously prolongs the action potential interval of the myocardial cells with the function of regulating potassium channel, and the efficacy is dose-dependent. The drug has no obvious effect on the action potential interval of the myocardial cells by regulating the potassium ion channel, when the drug concentration at 1 mg/ml in quadruplicate, there is no significant difference compared with the control group. The drug shows obvious effect on the action potential interval of the myocardial cells by regulating the potassium ion channel, when the drug concentration at 2 mg/ml in septuplicate, there is significant difference compared with the control group (P<0.05). The drug shows obvious effect on the action potential interval of the myocardial cells by regulating the potassium ion channel, when the drug concentration at 4 mg/ml in septuplicate, there is significant difference compared with the control group (P<0.05). The results are shown in FIGS. 2 and 3.