CN111514154A

CN111514154A - Application of notoginsenoside Fc in treating cerebral infarction

Info

Publication number: CN111514154A
Application number: CN202010022351.8A
Authority: CN
Inventors: 王峥涛; 高波; 杨莉; 吴晓俊; 张二云; 黄玲芳
Original assignee: Shanghai University of Traditional Chinese Medicine
Current assignee: Shanghai University of Traditional Chinese Medicine
Priority date: 2015-11-27
Filing date: 2015-11-27
Publication date: 2020-08-11
Also published as: CN106806378B; CN106806378A

Abstract

The invention relates to notoginsenoside F_CCan be used for preventing and treating cerebral infarction.

Description

Application of notoginsenoside Fc in treating cerebral infarction

Technical Field

The invention relates to the field of medicines, and in particular relates to a pharmaceutical application of notoginsenoside Fc.

Background

Thrombosis (thrombosis) refers to a pathological process in which under certain conditions, the formed components in blood form emboli in blood vessels (mostly small blood vessels), resulting in partial or complete blockage of blood vessels and blood supply disorder of corresponding tissues and organs. Depending on its composition, a thrombus can be divided into: platelet thrombus, erythrocyte thrombus, fibrin thrombus, mixed thrombus and the like. Depending on the type of blood vessel forming the thrombus, the thrombus can be classified into arterial thrombus, venous thrombus and capillary thrombus.

Thromboembolism (thrombo-embolism) refers to a pathological process in which a thrombus falls off from its formation site, partially or totally blocking certain blood vessels during the course of flowing with the blood, thereby leading to ischemia, hypoxia, necrosis (arterial thrombosis) and/or edema due to blood stasis (venous thrombosis) of the corresponding tissue organ.

The diseases caused by the above two pathological processes are generally called thrombotic diseases clinically. The etiology and pathogenesis of the disease are quite complex and are not completely clear so far, but the research in recent years shows that the occurrence and development of thrombotic diseases are mainly related to the following two factors:

first, the number of platelets is increased and the activity is enhanced. Various factors that increase the number and activity of platelets may induce and promote thrombotic diseases, such as thrombocythemia, accelerated platelet destruction caused by mechanical, chemical, biological and immunological reactions, etc. Platelet factors are now believed to play a more important role in the pathogenesis of arterial thrombosis (e.g., myocardial infarction).

Secondly, the blood coagulability is increased. Under various physiological and pathological conditions, the blood coagulation activity of human body can be obviously enhanced, which is represented by the increase of the level or activity of certain blood coagulation factors, such as stress reaction caused by pregnancy, old age, wound infection and the like, hyperlipidemia, malignant tumor and the like. The hypercoagulable state of blood is the basis of the onset of thrombotic disease.

Notoginsenoside fc (notogenoside fc) is a natural saponin compound, and is mainly present in notoginseng. Researches find that the notoginsenoside has the functions of reducing blood fat, resisting tumor, resisting oxidation and the like.

Disclosure of Invention

The invention aims to provide a new medical application of notoginsenoside Fc.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating vascular occlusive ischemia.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating vascular occlusive lower limb ischemia.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating or preventing thromboangiitis

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating vasculitis.

The invention provides application of notoginsenoside Fc in preparing a medicament for treating cerebral infarction.

The invention provides application of notoginsenoside Fc in preparing a medicament for treating acute cerebral infarction.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating or preventing myocardial ischemia or myocardial infarction.

Specifically, the invention provides an application of notoginsenoside Fc in preparation of a platelet aggregation inhibitor. The concentration of notoginsenoside Fc is 0.1-40 μ M, preferably 13.5 μ M.

In a second aspect, the invention provides an application of notoginsenoside Fc in preparing a blood coagulation inhibitor, wherein the concentration of notoginsenoside Fc is 1-240mg/ml, preferably 80 mg/ml.

In a third aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing thrombotic diseases, which comprises a therapeutically effective amount of notoginsenoside Fc, and the concentration of notoginsenoside Fc is 1-24mg/kg, preferably, 8.1 mg/kg. The thrombotic diseases include arterial thrombosis, venous thrombosis, and capillary thrombosis.

In a fourth aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing myocardial ischemia, comprising a therapeutically effective amount of notoginsenoside Fc, wherein the concentration of notoginsenoside Fc is 1-36mg/kg, preferably 12 mg/kg.

In a fifth aspect of the present invention, there is provided a pharmaceutical composition for treating or preventing myocardial infarction, comprising a therapeutically effective amount of notoginsenoside Fc, wherein the concentration of notoginsenoside Fc is 1-36mg/kg, preferably 12 mg/kg.

The fifth aspect of the invention provides application of notoginsenoside Fc in preparing a medicament for treating vascular occlusive lower limb ischemia, wherein the concentration of the notoginsenoside Fc is 0.1-240mg/ml, preferably 40 mg/ml.

The sixth aspect of the invention provides an application of notoginsenoside Fc in preparing a medicament for treating acute cerebral infarction, wherein the concentration of the notoginsenoside Fc is 0.1-240mg/ml, 1-36mg/kg, preferably 16 mg/kg.

Drawings

FIG. 1 Effect of notoginsenoside Fc on platelet aggregation Rate

FIG. 2 dose-effect relationship of notoginsenoside Fc on platelet aggregation rate

FIG. 3 Effect of Fc on the platelet aggregation Rate in Whole animals

FIG. 4 Effect of Fc on rat tail-broken clotting time

FIG. 5 Effect of Fc on rat activated partial thromboplastin time

FIG. 6 influence of Fc on four indices of human coagulation

FIG. 7 Effect of Fc on four indices of human coagulation

FIG. 8 Effect of Fc on the percentage of acute myocardial infarction area in rats

FIG. 9 pathological pictures of blank group of rat myocardial infarction operation

FIG. 10 pathological pictures of rat myocardial infarction sham operation group

FIG. 11 pathological picture of rat myocardial infarction positive drug group (4.1mg/kg)

FIG. 12 pathological picture of rat myocardial infarction administration group (4mg/kg)

FIG. 13 pathological picture of rat myocardial infarction administration group (12mg/kg)

Fig. 14 laser doppler blood flow profiles of Fc-treated group after the first day of surgery.

Fig. 15 graph of laser doppler blood flow changes in Fc-treated groups after the fourth day of surgery.

Fig. 16 laser doppler blood flow profiles of Fc-treated group after the eighth day of surgery.

Fig. 17 laser doppler blood flow profiles of Fc-treated group after the sixteenth day of surgery.

Fig. 18 twenty-sixth day after surgery, the operative limbs of the Fc-treated group had returned to 43% of the normal limbs.

FIG. 19 brain sections of Fc-treated group and control group were compared 2h after rat cerebral infarction.

Figure 20 percent infarct in Fc treated versus control groups.

Detailed Description

The invention arose in part from the unexpected discovery that: the notoginsenoside Fc can obviously inhibit the platelet aggregation of rats and prolong the time of activating partial thromboplastin. Therefore, notoginsenoside Fc is expected to be developed as a substance inhibiting platelet aggregation, i.e., a platelet aggregation inhibitor, and/or a substance inhibiting blood coagulation, i.e., a blood coagulation inhibitor, which may be in various forms including, but not limited to: drugs, health products, foods, daily necessities, and the like. These "platelet aggregation inhibitors" and "blood coagulation inhibitors" prepared from notoginsenoside Fc can be used for preventing and treating various thrombotic diseases such as arterial thrombosis, venous thrombosis and capillary thrombosis.

Furthermore, the invention provides application of notoginsenoside Fc in preparation of platelet aggregation inhibitors.

The invention also provides application of the notoginsenoside Fc in preparation of a blood coagulation inhibitor.

The invention also provides a pharmaceutical composition for treating thrombotic diseases, which comprises therapeutically effective amount of notoginsenoside Fc.

More preferably, the thrombotic disease is platelet aggregation hyperactivity or hypercoagulability.

The invention also provides application of the notoginsenoside Fc in preparing a pharmaceutical composition for treating vascular occlusive ischemia.

The invention also provides application of the notoginsenoside Fc in preparing a pharmaceutical composition for treating vascular occlusive lower limb ischemia.

The invention also provides application of the notoginsenoside Fc in preparing a pharmaceutical composition for treating vasculitis.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating or preventing thromboangiitis obliterans.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating or preventing cerebral infarction.

The invention provides application of notoginsenoside Fc in preparing a pharmaceutical composition for treating or preventing acute cerebral infarction.

The molecular formula of notoginsenoside Fc (Notogenoside Fc) is as follows: C58H98O26, molecular weight: 1210.63, having the formula:

R＝Xyl1-6Glc；R1＝Xyl1-2Glc1-2Glc

notoginsenoside Fc of the present invention can be commercially obtained from Sigma chemical company, kywman scientific and biological limited, or isolated from notoginsenoside using a conventional method in the art. The purity of the product meets the pharmaceutical standard.

The notoginsenoside Fc of the invention is prepared into a medicament as an example. The notoginsenoside Fc of the present invention can be used alone or in the form of a pharmaceutical composition. The pharmaceutical composition comprises notoginsenoside Fc as an active ingredient and a pharmaceutically acceptable carrier. Preferably, the pharmaceutical composition of the present invention contains 0.1 to 99.9% by weight of notoginsenoside Fc of the present invention as an active ingredient. The pharmaceutical carrier does not damage the pharmaceutical activity of the notoginsenoside Fc, and the effective dosage of the notoginsenoside Fc is nontoxic to human bodies.

Such pharmaceutically acceptable carriers include, but are not limited to: lecithin, aluminum stearate, alumina, ion exchange materials, self-emulsifying drug delivery systems, tweens or other surfactants, serum proteins, buffer substances such as phosphates, glycine, sorbic acid, water, salts, electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, magnesium silicate, mixtures of saturated fatty acid partial glycerides, and the like.

Other conventional pharmaceutical adjuvants such as binder (e.g. microcrystalline cellulose), filler (e.g. starch, glucose, anhydrous lactose and lactose beads), disintegrant (e.g. crosslinked PVP, croscarmellose sodium, low-substituted hydroxypropylcellulose), lubricant (e.g. magnesium stearate), and absorption enhancer, adsorption carrier, flavoring agent, sweetening agent, excipient, diluent, wetting agent, etc.

The notoginsenoside Fc and pharmaceutical compositions thereof of the present invention can be prepared according to conventional methods in the art and can be administered by enteral or parenteral or topical routes. The oral preparation comprises capsule, tablet, oral liquid, granule, pill, powder, pellet, and unguent; parenteral preparations include injections and the like; topical preparations include creams, patches, ointments, sprays, and the like. Oral formulations are preferred.

The administration route of the notoginsenoside Fc and the pharmaceutical composition thereof can be oral, sublingual, transdermal, intramuscular or subcutaneous, skin mucosa, vein, urethra, vagina and the like.

Besides preparing medicines, various food additives such as antioxidants, pigments, enzyme preparations and the like can be added into the notoginsenoside Fc to prepare health-care food according to the conventional method in the field.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. All percentages, ratios, proportions, or parts are by weight unless otherwise specified.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

The features mentioned above with reference to the invention, or the features mentioned with reference to the embodiments, can be combined arbitrarily. All the features disclosed in this specification may be combined in any combination and each feature disclosed in this specification may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.

Example 1:

the notoginsenoside Fc used in the present example was isolated and prepared by the Chinese medicine institute of Shanghai medical university (clopidogrel purchased from China biological products institute). The purity of the samples used all met the pharmaceutical standards.

1. Experimental Material

1.1 medicinal materials

Notoginsenoside Fc (molecular weight 1211.38, HPLC purity greater than or equal to 99%, Chinese medicine research institute); clopidogrel (molecular weight 419.90, HPLC purity more than or equal to 98%, purchased from China Biometrics institute); human standard plasma, four test reagents for clotting (all available from SIEMENS, germany); adenosine Diphosphate (ADP) (from Sigma Co.)

1.2 Experimental animals

SD rats weighing 230 ± 20g, provided by the experimental animals center of the university of medicine in shanghai, animal certification No.: SYXK (Shanghai) 2008-0016. Placing in conventional breeding environment, and freely taking food and drinking water.

2. Experimental methods

2.1 Effect of Fc on platelet aggregation

2.1.1 in vitro platelet aggregation assay: adding sodium citrate into the blood to anticoagulate. Centrifuging at 100g for 5min, and collecting supernatant to obtain Platelet Rich Protein (PRP); adding the obtained PRP to prostacyclin (2. mu.g/ml), and centrifuging for 5min (700 g); then adding 0.9% of normal saline to clean twice; after washing, the platelets were suspended in Tyrode buffered at a concentration of 4 x 108 for future use; platelet aggregation assay by turbidimetry: platelet plasma was placed in a cuvette and a inducer (ADP: 5-adenosine diphosphate disodium salt: 1X 10-4mol/L) was added. Then, the small magnetic particles are used for stirring, so that the blood platelets gradually aggregate, the turbidity of the blood plasma is reduced, and the transmittance is increased. This change was recorded, forming a dynamic curve of platelet aggregation. The platelet aggregation curve was automatically measured, recorded, and plotted by a platelet aggregation meter with the aggregation rate and transmittance of PRP being 0 and the aggregation rate and transmittance measured from the suspension being 100%. The judgment of the inhibition or promotion of aggregation is mainly to observe whether the synergistic effect or the antagonistic effect occurs with the inducer. The synergistic effect is to promote the platelet aggregation, and the antagonistic effect is to inhibit the platelet aggregation.

2.1.2 Whole animal platelet aggregation assay: platelets in the arterial blood stream adhere to the thread as the rough surface of the thread contacts, forming platelet thrombi on the surface. When the adhesion-aggregation function of platelets is inhibited, the thrombus is light in weight. Therefore, the function of platelet adhesion and aggregation can be determined from the thrombus weight.

2.1.2 evaluation of Whole animal blood clotting function

2.1.2.1 measurement of clotting time of rat tail broken: crossing the tail tip of the rat at a position of 0.5cm by using a pair of scissors, and sucking off blood drops for 1 time by using filter paper every 30s when the blood begins to be recorded after the blood automatically overflows until the blood flow naturally stops (no blood exists when the filter paper sucks), namely the bleeding time.

2.1.2.2 measurement of four coagulation indexes: 1) four indices of blood coagulation: prothrombin Time (PT): mainly reflects the condition of the intrinsic coagulation system and is commonly used for monitoring the dosage of heparin. The increase is seen in reduced levels of plasma factor viii, factor ix and factor XI: such as hemophilia a, hemophilia B, and factor XI deficiency; the decrease is seen in the hypercoagulable state: such as the increased activity of procoagulant substances entering the blood and coagulation factors; activated Partial Thromboplastin Time (APTT): mainly reflects the condition of the intrinsic coagulation system and is commonly used for monitoring the dosage of heparin. The increase is seen in reduced levels of plasma factor viii, factor ix and factor XI: such as hemophilia a, hemophilia B, and factor XI deficiency; the decrease is seen in the hypercoagulable state: such as the increased activity of procoagulant substances entering the blood and coagulation factors; thrombin time assay (thrombin, TT): mainly reflecting the time of conversion of fibrinogen to fibrin. The increase is seen in DIC hyperfibrinolysis stage, hypo (no) fibrinogenemia, hemoglobinemia, and increase in fibrin (ogen) degradation products (FDPs) in blood; the reduction has no clinical significance; fibrinogen (Fibrinogen, FIB): mainly reflecting the fibrinogen content. The increase is seen in acute myocardial infarction, and the decrease is seen in DIC consumption hypo-fibrinolysis, primary fibrinolysis, severe hepatitis, and liver cirrhosis;

2.2 methods of grouping and administering drugs

2.2.1 platelet aggregation assay design:

blank control: 0.9% physiological saline (NS).

Inducer group: adenosine Diphosphate (ADP) (10mM)

Positive control drug: clopidogrel (5 mM).

The medicine group is as follows: six of each group, Fc was dissolved in double distilled water (56 μ M).

2.2.2 in vivo platelet aggregation function experiment

Blank control: 6, 0.9% physiological saline (NS).

Positive control drug: 6, clopidogrel (13.4 mg/kg).

The medicine group is as follows: fc was dissolved in double distilled water (8.1mg/kg) for 6 cells.

2.2.3 Experimental design of coagulation time of rat tail

Blank control: 6, 0.9% physiological saline (NS).

Positive control drug: 6, clopidogrel (intravenous injection 13.4 mg/kg).

The medicine group is as follows: fc was dissolved in double distilled water (8.1mg/kg for intravenous injection) for 6 cells.

2.2.4 assay design for four items of coagulation

Blank control: 6, 0.9% physiological saline (NS).

Positive control drug: 6, clopidogrel (160. mu.g/ml).

The medicine group is as follows: fc was dissolved in double distilled water (80. mu.g/ml) for 6 cells.

2.4 statistical analysis

All experimental data were repeated 3 times, the results were expressed as mean ± standard deviation, and One-way analysis of variance (One-way ANOVA) and LSD test were performed on the experimental data using SPSS 13.0 statistical software, with P <0.05 being statistically significantly different.

3. Results

3.1 inhibitory action of notoginsenoside Fc on platelet aggregation.

3.1.1 Effect of Fc on platelet aggregation Rate

Fig. 1 shows the effect of notoginsenoside Fc on platelet aggregation rate, and it can be seen from fig. 1 that: the aggregation rate of the inducer (ADP) was 52.9% in the single group, 25.8% after co-administration of Fc (both administered at 200. mu.M), and the aggregation rate of the Fc group was smaller than that of the inducer (P <0.05) alone and smaller than that of the positive drug group (P < 0.05). This indicates platelet aggregation induced by the Fc antagonist inducer.

3.1.1 dose-effect relationship of Fc to platelet aggregation Rate

Fig. 2 shows the dose-effect relationship of notoginsenoside Fc to platelet aggregation rate, and it can be seen from fig. 2 that: the dose-effect relationship of Fc (0.1. mu.M, 10. mu.M, 20. mu.M, 30. mu.M, 40. mu.M) on the platelet aggregation inhibition effect was that the platelet aggregation rate gradually decreased from the dose range of 0.1. mu.M to 40. mu.M. The aggregation rate decreases in turn with increasing dose. The aggregation rate is reduced from 47.9 percent to 28.1 percent, and the reduction amplitude reaches 19.8 percent. The IC50 value was 13.5. mu.M.

3.1.3 Effect of Fc on the platelet aggregation Rate of Whole animals

Fig. 3 shows the effect of Fc on the platelet aggregation rate in whole animals, as can be seen from fig. 3: influence of in vivo platelet aggregation function of notoginsenoside Fc. By weighing the wet heavy thrombus we found: fc still has strong activity in the body case. We used the intravenous method to administer Fc: 8.1 mg/kg. Five minutes after administration, the collateral circulation in vitro was started, fifteen minutes after circulation, the circulation was terminated, and the thrombus was taken out. Blank control group was significantly different from Fc dosed group (. about.p < 0.01).

3.2 Effect of notoginsenoside Fc on blood coagulation function

3.2.1Fc determination of coagulation time of rat tail-broken

FIG. 4 shows the effect of Fc on coagulation time of rat tail-cleft, as can be seen from FIG. 4: fc has obvious influence on the tail breaking bleeding time of rats. Thrombus weight of Fc 28.2 mg; significantly less than blank: 37.2mg (. times.p < 0.01).

3.3 Effect of Fc on four indices of blood coagulation in rats and humans

FIG. 5 shows the effect of Fc on the Activated Partial Thromboplastin Time (APTT) in rats, as can be seen in FIG. 5: the dose-effect relationship of Fc to APTT is five dose groups of 5 μ g/ml, 10 μ g/ml, 20 μ g/ml, 80 μ g/ml and 160 μ g/ml. Different doses showed a gradual increase in effect, with a dose of 5 μ g/ml APTT time of Ft 1: 19 seconds, 240. mu.g/ml 32 seconds. The amplitude of variation was 13 seconds.

FIG. 6 shows the effect of Fc on four indices of human coagulation, as can be seen from FIG. 6: fc significantly extended APTT time, reaching 126 seconds at the 80 μ g/ml dose level, significantly above the blank (91 seconds) (. x.p < 0.01). Fc at a dose level of 240 μ g/ml, APTT time was only 82 seconds, significantly lower than blank (91 seconds) (. x.p < 0.01).

Figure 7 demonstrates the effect of Fc on human PT time: PT was also longer than blank (. < 0.05); . PT time was 17 seconds, significantly lower than blank (20 seconds) (. P < 0.05).

Example 2: reducing effect of notoginsenoside Fc on myocardial infarction area of operation rat and anti-myocardial ischemia effect

First, experimental material

1.1 medicinal materials

Notoginsenoside Fc (molecular weight 1211.38, HPLC purity greater than or equal to 99%, Chinese medicine research institute); clopidogrel (molecular weight 419.90, HPLC purity more than or equal to 98%, purchased from China Biometrics institute);

1.2 Experimental animals

SD rats weighing 320 ± 20g, provided by the experimental animals center of the university of medicine in shanghai, animal certification No.: SYXK (Shanghai) 2008-0016. Placing in conventional breeding environment, and freely taking food and drinking water.

Secondly, the method comprises the following steps:

the model making method comprises the following steps: SD rats 30 (provided by the animal experiment center of the university of medicine in shanghai), body weight: 320-370 g. Rat pentobarbital (35mg/kg) was anesthetized and fixed supine; cutting the center of the neck, separating trachea, performing tracheotomy, inserting a respiratory tube, and adjusting the frequency and tidal volume of a respirator; the chest (3-4 intercostals) is opened through the median incision of the sternum, a thin operation needle and a six-gauge thread are inserted into the left auricle and then the pulmonary artery is ligated out of the cone. Judging success of branch descending before suture

The standard adopts visual observation (the blood supply area of the anterior descending branch is dark or pale), so that the chest is closed rapidly. The sham operation group only threaded around the anterior descending branch of the left coronary artery without ligation, the same as the operation group.

TTC dyeing: freezing and storing at the temperature of minus 20 ℃, taking out until the heart is frozen hard, and cutting the heart into 5-6 slices along the long axis of the left ventricle, wherein each slice is 3-4 mm thick. Then placing the mixture into 0.5% TTC solution, incubating the mixture for 15min at 37 ℃, taking out the myocardial slices and fixing the myocardial slices in 4% formaldehyde solution.

Index collection: the myocardial tablets are in two different colors, and the red is normal myocardial tissue; white or grayish black is necrotic myocardial tissue. The myocardium was cut in two different colors and then dried with filter paper, and the weight was weighed. The area of myocardial ischemia (%) — total heart weight ÷ infarcted myocardium weight × 100% was calculated as follows. Meanwhile, a part of the heart is collected for pathological section analysis.

Third, design of experiment

2.2.5 Experimental design for acute myocardial infarction of rat

Surgical blank group: 6 patients were administered with 0.9% Normal Saline (NS) after surgery.

The sham operation group: 6, open the chest, thread but not ligate.

A positive drug group: 6 patients were administered ticagrelor (4.1mg/kg) post-operatively.

4mg/kg Fc administration group: 6 were administered Fc (4mg/kg) post-operatively.

12mg/kg Fc administration group: 6 were administered Fc (12mg/kg) post-operatively.

Fourthly, conclusion:

FIG. 8 shows that Fc has the effect of obviously reducing the myocardial infarction area percentage of the operation rats. Moreover, the 4mg/kg administration group and the 12mg/kg administration group have obvious dose-effect relationship (P < 0.001). The curative effect of the 12mg/kg administration group is better than that of the positive medicine group (4.1mg/kg) (P is less than 0.01). Each administration group was significantly different from the control group.

FIG. 9: the surgical blank group indicated: the arrangement of the myocardial cells is disordered, the cytoplasm of the cells is heavily infected with acidophilic, part of the cytoplasm is less, part of the cytoplasm is increased, the cell nucleus is unevenly distributed, the formation of multi-core cells can be locally seen, obvious vacuole degeneration can be seen in the stroma, and inflammatory cells are rare.

FIG. 10: surface of the sham-operated group: the arrangement of the myocardial cells is normal, the cytoplasm of the cells is rich and acidophilic, the sizes of the cell nucleuses are consistent and uniformly distributed, and inflammatory cells are not seen in the interstitium.

FIG. 11: the positive medicine group shows that: the arrangement of the myocardial cells is locally disordered, the cytoplasm is acidophilic, the myocardial cells are rich, the sizes of the nuclei are consistent, the distribution is uneven, multinucleated cells are seen, the interstitial vacuole degeneration is obvious, and inflammatory cells are rare.

FIG. 12: the group administered 4mg/kg indicated: the arrangement of the cardiac muscle cells is approximately normal, the cytoplasm of the cells is acidophilic, local cell nucleuses are gathered, a small amount of inflammatory cell infiltration can be seen in the interstitium, and vacuole degeneration is obvious.

FIG. 13: the group administered 12mg/kg indicated: the arrangement of the myocardial cells is approximately normal, the local cell cytoplasm is broken, the cell cytoplasm is rich, the cell nucleus is increased, the interstitial vacuole degeneration is reduced, and the inflammatory cell infiltration is obvious.

Example 3: therapeutic action of notoginsenoside Fc on mouse vascular occlusive lower limb ischemia

First, experimental material

1.1 medicinal materials

Notoginsenoside Fc (molecular weight 1211.38, HPLC purity greater than or equal to 99%, Chinese medicine research institute); a positive drug (clopidogrel: molecular weight 419.90, HPLC purity more than or equal to 98%, purchased from China Biochemical institute); notoginseng radix total saponin (from Chinese medicinal materials);

1.2 Experimental animals

C57 mice weighing 20 ± 5g, provided by the experimental animals center of the university of medicine in shanghai, animal certification numbers: SYXK (Shanghai) 2008-0016. Placing in conventional breeding environment, and freely taking food and drinking water.

Secondly, the method comprises the following steps:

the model making method comprises the following steps: an autoclave before an operation sterilizes surgical instruments. Mice are anesthetized with isoflurane or other anesthetic until it is unresponsive to external stimuli.

Then placed in front of the upper operating table in a supine position. The hair was thoroughly removed using an electric razor and depilatory cream. The mouse is placed on the operating table in supine position (preferably with a heating pad to keep the sleeping mouse warm), the back foot is fixed with tape, and the skin is scrubbed with iodine and alcohol. The skin was incised approximately 1 cm long from the knee to the inside of the thigh using fine forceps and surgical scissors. The tip of the cotton swab was soaked with PBS and the subcutaneous fat was brushed gently. Next, a cauterization or blunt needle is applied to poke open the soft tissue, exposing the underlying femoral artery. The drag hook is used for pulling open the wound to better expose the blood vessel of the lower limb. The exposed neurovascular bundle was gently stripped. Then, the thin tip of the cotton swab was soaked with a fine forceps and PBS, and the femoral artery, femoral vein and nerve were dissected and separated. After stripping, performing surgical ligation on femoral artery at a position near the groin by using 7-0 silk threads, and performing secondary ligation at a position near the femoral artery; the same action is done near the knee. All arteries connected to the femoral artery (approximately three to four) were tied with 5-0 wire loop details. The femoral vein is resected to the femoral artery near the knee. The draw hook was removed and the incision closed with 5-0 sutures. The mouse is placed on top of the heating pad and monitored until awake.

Index collection: and adopting moorLDLS laser doppler to collect blood flow change data of the mice in different time periods after the operation.

Third, design of experiment

Surgical blank group: 6 patients were intraperitoneally injected (ip) with 0.9% Normal Saline (NS) after surgery.

A positive drug group: 6 patients were intraperitoneally injected with clopidogrel (ip) (10mg/kg) after surgery.

Fc administration group: 6 patients were intraperitoneally injected (ip) with notoginsenoside Fc (40mg/kg) after surgery.

Panax Notoginseng Saponins (PNS): 6, intraperitoneal (ip) PNS (50mg/kg) after surgery

Fourthly, conclusion:

fig. 14 laser doppler blood flow profiles of Fc-treated group after the first day of surgery. The blood flow is small because the blood vessel is cut off.

Fig. 15 graph of laser doppler blood flow changes in Fc-treated groups after the fourth day of surgery. The blood flow was greater than on the first day.

Fig. 16 laser doppler blood flow profiles of Fc-treated group after the eighth day of surgery. Blood flow was already significantly greater than on the first day. The non-blood flow area is reduced, and the blood flow of the ischemic limb is further recovered.

Fig. 17 laser doppler blood flow profiles of Fc-treated group after the sixteenth day of surgery. The ischemic limb blood flow had significantly recovered.

FIG. 18 twenty-sixth day after surgery (see Table 1), operative limbs in the Fc-treated group had returned to 43% of normal limbs, which was significantly better than 25% in the model group (P <0.05), better than 27% of the positive drug (P <0.05), and better than total arasaponin (PNS) (23%) (P <0.05) which is currently widely used.

Table 1.The data of Fc treated group(after 28days)

Example 4: therapeutic effect of notoginsenoside Fc on acute cerebral infarction model infarction of rat

First, experimental material

1.1 medicinal materials

Notoginsenoside Fc (molecular weight 1211.38, HPLC purity greater than or equal to 99%, Chinese medicine research institute);

1.2 Experimental animals

SD rats weighing 200 ± 50g, provided by the experimental animals center of the university of medicine in shanghai, animal certification No.: SYXK (Shanghai) 2008-0016. Placing in conventional breeding environment, and freely taking food and drinking water.

Secondly, the method comprises the following steps:

the model making method comprises the following steps: an autoclave before an operation sterilizes surgical instruments. 1% of the pentobarbital anesthetized mice (40mg/kg) until it was unresponsive to external stimuli.

Manufacturing a wire embolism cerebral infarction model: the skin was incised along the middle of the meridian. The subcutaneous tissue was isolated blunt. After separating the anterior muscle of the trachea, the anterior muscle of the trachea is separated downwards along the tendon of the sternocleidomastoid tendon at the right side, and the retractor can be pulled up after the carotid sheath is seen. The arterial sheath was isolated and the smooth common carotid artery was visualized after isolation. Separating the common carotid artery, the external carotid artery and the internal carotid artery, and ligating the common carotid artery and the external carotid artery. The internal carotid artery is closed and the common carotid artery is cut into a small opening by an ophthalmic scissors. Inserting a fishing line: the selection of the fish line in the step is key, and the fish line is selected according to the weight of the rat. The line of 0.26mm below 250g, 0.26mm dipped wax or 0.28mm dipped wax of 300g may be used. The fish line dipped with wax is used as a model, and the advantages of good control of the incoming line depth and difficult subarachnoid hemorrhage are achieved. After successful ligation of the middle line, the first marker was seen to be about 2mm from the arterial bifurcation. After sewing, the skin suture is sterilized by alcohol cotton ball. The rats were returned to their cages. The brains were sacrificed after 2 h. Freezing brain at-20 deg.C for 30min, and slicing (5-6 slices). After placing into TTC (0.5%) for 30min, the peduncle area is white, and the normal area is red. The ratio (%) of the area of the infarcted area to the area of the whole brain piece is the percentage of infarct in the brain. For brain area measurement, Image J1.4 g was used.

Third, design of experiment

Surgical blank group: 3, post-operative intravenous (iv) 0.9% Normal Saline (NS).

Sanchinoside Fc administration group: (iii) 3 patients, and (iv) notoginsenoside Fc (16mg/kg) was injected intravenously after surgery.

Fourthly, conclusion:

Figure 20 percent infarct in Fc treated versus control groups. The infarct size of the Fc-treated brains was significantly reduced compared to the control group (p < 0.0001)

Table 1: percent (%) data of cerebral infarction between surgery blank group and Fc-treated group (n ═ 3)

The various aspects of the invention are addressed above. It should be understood, however, that equivalent changes and modifications may be made thereto by those skilled in the art without departing from the spirit of the present invention, and that such changes and modifications are intended to be covered by the appended claims.

Claims

1. Application of notoginsenoside Fc in preparing pharmaceutical composition for treating cerebral infarction is provided.

2. Application of notoginsenoside Fc in preparing pharmaceutical composition for treating acute cerebral infarction is provided.

3. An application of notoginsenoside Fc in preparing the medicine for treating cerebral infarction features that the concentration of notoginsenoside Fc is 0.1-240 mg/ml.

4. Use of notoginsenoside Fc according to claims 1-6 for the preparation of a pharmaceutical composition, characterized in that the pharmaceutical composition can be prepared according to methods conventional in the art and can be administered by enteral or parenteral or topical route; the oral preparation comprises capsule, tablet, oral liquid, granule, pill, powder, pellet, and unguent; parenteral preparations include injections and the like; topical preparations include creams, patches, ointments, sprays, and the like; the health food is prepared by adopting antioxidant, pigment and enzyme preparation according to the conventional method in the field.

5. Use of notoginsenoside Fc according to claims 1-6 for the preparation of a pharmaceutical composition, characterized in that the route of administration can be oral, sublingual, transdermal, intramuscular or subcutaneous, mucocutaneous, intravenous, urethral, vaginal, etc.