CN104814976A - Novel application of acanthopanax polysaccharide - Google Patents
Novel application of acanthopanax polysaccharide Download PDFInfo
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- CN104814976A CN104814976A CN201510255886.9A CN201510255886A CN104814976A CN 104814976 A CN104814976 A CN 104814976A CN 201510255886 A CN201510255886 A CN 201510255886A CN 104814976 A CN104814976 A CN 104814976A
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Abstract
The invention discloses novel application of an acanthopanax polysaccharide. The acanthopanax polysaccharide is applied to preparation of a drug for treating immunological liver injury. The acanthopanax polysaccharide ASPA has certain effects on preventing and protecting immunological liver injury, the protecting effect of the polysaccharide is directly embodied on the aspects of protecting the liver and lowering transaminase, in addition, the activities of SOD and GSH-Px can also be improved through inhibiting MDA, so that the oxidative damage resistance of hepatic cells can be improved. The acanthopanax polysaccharide is applied to the field of pharmaceuticals.
Description
Technical field
The present invention relates to a kind of novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
Background technology
Immunologic liver injury (Immunological Liver Injury, ILI) refer to because immune response causes, to discharge various free radical, cytokine and inflammatory mediator cause the formation of inflammatory cell infiltration in hepatic tissue, inflammatory granuloma, hepatic cords structural deterioration is the inflammatory and immune response of feature, finally cause the hepatocyte injury of hepatic necrosis and apoptosis.Immunologic mechanism serves crucial effect in course of liver damage, is viral hepatitis, one of liver cirrhosis, the finally important pathogenesis developing into hepatocarcinoma, and determines lapsing to or developing of disease.The main means of prevention of immunologic liver injury is embodied in three aspects: the application of (1) antiviral therapy medicine, as interferon, ucleosides analog lamivudine etc.; (2) immunomodulator, as non-specific immunomodulator such as thymosins; (3) the liver protecting and ALT lowering, fibrosis medicine, as interferon, colchicine etc.The untoward reaction that nucleoside analog causes has myopathy, nephrotoxicity, peripheral neuropathy, lactic acidosis etc.; Interferon prolonged application can produce cardiac toxicity; Colchicine also has severe toxicity, and common have nausea,vomiting,diarrhea gastrointestinal reaction and the kidney damage such as hematuria, oliguria and have direct repression to cause the side effect such as aplastic anemia to bone marrow.The medicine of common treatment immunologic liver injury is as bifendate drop pill etc., and long-term taking can cause hepatocellular damage, causes acute hepatic failure, acute hepatitis, Cholestatic hepatitis, fatty liver, the side effect such as blood vessel injury.
Summary of the invention
The invention provides the novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide.
The novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide of the present invention refers to the medicine of Radix Et Caulis Acanthopanacis Senticosi polysaccharide for the preparation for the treatment of immunologic liver injury.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide Acanathopanax Senticosus Polysaccharides (ASPS) is the vegetable polysaccharides extracted from studies on plants of Acanthopanax Miq. in Araliaceae Radix Et Caulis Acanthopanacis Senticosi Acanthopanax Senticosus root and rhizome.Be the macromolecular substances that a class has natural attribute, being a class by aldose or ketose by the multi-form natural polymers be formed by connecting of glycosidic bond, is one of main active of Radix Et Caulis Acanthopanacis Senticosi.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide of the present invention is in the application for the treatment of immunologic liver injury, find that Radix Et Caulis Acanthopanacis Senticosi polysaccharide ASPS has certain prevention and protective effect to immunologic liver injury, polysaccharide is not only embodied directly in the liver protecting and ALT lowering aspect to the protective effect of hepatic injury, the activity of SOD and GSH-Px can also be improved from the ability improving hepatocellular anti-oxidative damage by suppressing MDA, its mechanism may remove body free radical with him, enhancing body anti peroxidation of lipid ability is relevant with the expression of adhesion factor with reduction inflammatory cytokine, in addition, Radix Et Caulis Acanthopanacis Senticosi polysaccharide has safety and stability, Mutiple Targets, the advantage of hypotoxicity etc., that it has very large advantages and potential on treatment immunologic liver injury.
Accompanying drawing explanation
Fig. 1 is the flow chart extracting Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1;
Fig. 2 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury model control group hepatic pathology;
Fig. 3 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury blank group hepatic pathology;
Fig. 4 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury positive drug group hepatic pathology;
Fig. 5 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury inhibitor group hepatic pathology;
Fig. 6 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury ASP high dose group hepatic pathology;
Fig. 7 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to dosage group hepatic pathology in Immune liver injury ASP;
Fig. 8 is that the Radix Et Caulis Acanthopanacis Senticosi polysaccharide in test 1 damages 40 times of light microscopic figure to Immune liver injury ASP low dose group hepatic pathology;
Fig. 9 is that test 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide is to the effect diagram of Immune liver injury serum transaminase; Wherein # represents comparing of blank group and model group, and * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.05; ## represents comparing of blank group and model group, p<0.01;
for ALT concentration U/L,
for AST concentration U/L;
Figure 10 is the effect diagram of test 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide to Immune liver injury serum IL-2, IL-4, IFN-γ; Wherein * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.05; ## represents comparing of blank group and model group, and * * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.01;
for IL-2 concentration pg/mL,
for IL-4 concentration pg/mL,
for IFN-γ concentration ng/L;
Figure 11 is that test 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide is to GSH-PX, MDA, SOD effect diagram in Immune liver injury murine liver tissue; Wherein * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.05; ## represents comparing of blank group and model group, and * * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.01;
for GSH-PX unit of activity,
for MDA content nmol/mL, ■ are total SOD vigor U/mgprot;
Figure 12 is that test 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide is to IL-1 β, NO, TNF-α effect diagram in Immune liver injury murine liver tissue; Wherein * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.05; ## represents comparing of blank group and model group, and * * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.01;
for IL-1 β concentration ng/L,
for NO concentration μm ol/L,
for TNF-α ng/L;
Figure 13 is that test 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide is to IL-2, IL-4 and INF-γ effect diagram in Immune liver injury mouse boosting tissue; Wherein * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.05; ## represents comparing of blank group and model group, and * * represents that dosage group in ASPS, ASPS high dose group, bifendate positive drug control group, PDTC inhibitor matched group compare with model group, p<0.01;
for IL-2 concentration pg/m,
for IL-4 concentration pg/m,
for INF-γ concentration ng/L.
Detailed description of the invention
Technical solution of the present invention is not limited to following cited detailed description of the invention, also comprises the combination in any between each detailed description of the invention.
Detailed description of the invention one: the novelty teabag of present embodiment Radix Et Caulis Acanthopanacis Senticosi polysaccharide refers to the medicine of Radix Et Caulis Acanthopanacis Senticosi polysaccharide for the preparation for the treatment of immunologic liver injury.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide Acanathopanax Senticosus Polysaccharides (ASPS) is the vegetable polysaccharides extracted from studies on plants of Acanthopanax Miq. in Araliaceae Radix Et Caulis Acanthopanacis Senticosi Acanthopanax Senticosus root and rhizome.Be the macromolecular substances that a class has natural attribute, being a class by aldose or ketose by the multi-form natural polymers be formed by connecting of glycosidic bond, is one of main active of Radix Et Caulis Acanthopanacis Senticosi.
Radix Et Caulis Acanthopanacis Senticosi polysaccharide of the present invention is in the application for the treatment of immunologic liver injury; find that Radix Et Caulis Acanthopanacis Senticosi polysaccharide ASPS has certain prevention and protective effect to immunologic liver injury; polysaccharide is not only embodied directly in the liver protecting and ALT lowering aspect to the protective effect of hepatic injury; the activity of SOD and GSH-Px can also be improved from the ability improving hepatocellular anti-oxidative damage by suppressing MDA; its mechanism may remove body free radical with him, and enhancing body anti peroxidation of lipid ability is relevant with the expression of adhesion factor with reduction inflammatory cytokine.In addition, Radix Et Caulis Acanthopanacis Senticosi polysaccharide has the advantage of safety and stability, Mutiple Targets, hypotoxicity etc., is that it has very large advantages and potential on treatment immunologic liver injury.
Detailed description of the invention two: present embodiment and detailed description of the invention one are 15.93mg/kg/d unlike the dosage of described medicine.Other is identical with detailed description of the invention one.
Detailed description of the invention three: present embodiment and detailed description of the invention one or two unlike: described pharmaceutical administration is oral.Other are identical with detailed description of the invention one or two.
For verifying beneficial effect of the present invention, carry out following test:
Test 1:
The novelty teabag of this test Radix Et Caulis Acanthopanacis Senticosi polysaccharide refers to the medicine of Radix Et Caulis Acanthopanacis Senticosi polysaccharide for the preparation for the treatment of immunologic liver injury.
The test of Radix Et Caulis Acanthopanacis Senticosi polysaccharide treatment immunologic liver injury is as follows:
This test Radix Et Caulis Acanthopanacis Senticosi polysaccharide adopts water extract-alcohol precipitation method to extract, and extracts flow chart as shown in Figure 1.
1, the foundation of animal model
Adopt ConA inducing mouse immunologic liver injury, mice last gavage 0.5h, tail vein injection 20mg/kg ConA is with induced liver injury, and blank group injection Isodose normal saline, puts to death animal and draw materials, detect indices after 8h.
2, animal grouping and process
Experimental animal is divided into 7 groups at random, blank group, model control group, ASPS low dose group (36.25mg/kg/d), dosage group (72.5mg/kg/d) in ASPS, ASPS high dose group (145mg/kg/d), bifendate positive drug control group, PDTC inhibitor matched group, often organizes 10, male and female half and half.Blank group is with 0.1ml/10g 0.9%NaCl solution gastric infusion, and administration group presses high, medium and low three dosage difference gavage 1 time, continuous 7d every day.Positive drug group (200mg/kg/d) gastric infusion, once a day, continuous 7d.The normal saline of inhibitor matched group gavage every day Isodose, 0.5h, 200mg/kg lumbar injection PDTC before modeling.Equally ConA and PDTC is diluted for 0.2ml volume.
3, test is drawn materials
Modeling terminates rear mice fasting about 8h, and posterior orbit of weighing gets blood, and the centrifugal 10min of 3500rh, prepares serum, is stored in-20 DEG C of refrigerators and detects next day.Then mice takes off vertebra execution, gets liver and spleen, and liver specimen is fixing to prepare pathological section with 10% formalin, gets liver and is about 0.5g, float blood with 4 DEG C of normal saline flushings, be made into 10% liver tissue homogenate's liquid with methanol, the centrifugal 15min of 3500rh, get supernatant, 4 DEG C frozen, in order to detecting.Get spleen and be about 0.05g, with legal system for tissue homogenate.Residue liver, spleen tissue are dipped in rapidly in liquid nitrogen frozen, give over to Western blot and detect and PCR detection.
4, test method and result
4.1 liver histopathology inspections
Put to death animal, take out liver, rinse floating blood with cold saline, wipe dry.Get 1.5cm × 1.5cm × 0.4cm in hepatic portal place and right lobe of the liver, fix by 10% neutral formalin immediately.90% ethanol dehydration, dimethylbenzene is transparent, paraffin embedding, and microtome does 4 μm of sections, and conventional dewaxing, HE dyes, neutral gum mounting.With microscopic examination, and PM-20 type microscope camera system is taken a picture, and sees Fig. 2 ~ Fig. 8, from Fig. 2 ~ Fig. 8, the even red dye of light Microscopic observation normal group liver HE dyeing liver cytoplasm, nucleus normal in size, the light dye of caryoplasm, lobules of liver clear in structure, hepatic cords marshalling.Have a large amount of lymphocytic infiltration in the visible liver of model group, particularly evident in portal area, hepatic cords arrangement destroys, and liver cell endochylema loosens swelling, and liver cell nuclear swelling is broken, visible obviously point-like or lamellar necrotic area, the dense dye of liver cell nuclear pyknosis.The pathological change compared with model group of ASPS high dose group obviously alleviates, between inhibitor group and positive drug group two groups; Tissue pathologies change's no significant difference between dosage group in ASPS low dose group and ASPS, but degree of injury is greater than inhibitor group, inhibitor group degree of injury is in ASPS between dosage group and positive drug group (n=10).
To the classification of hepatic tissue pathology degree of injury, the results are shown in Table 1; Wherein "-", obvious pathological change is had no; "+", the swelling of part hepatic tissue is with the spotty necrosis be dispersed in, and there is a little inflammatory cell infiltration portal area; " ++ ", swelling of liver cell has spotty necrosis and collections necrosis, the Granuloma formation be dispersed in as seen, and there is massive inflammatory cells infiltrated portal area; " +++ ", swelling of liver cell, has large lamellar downright bad, has Granuloma formation, and portal area and surrounding have massive inflammatory cells infiltrated.
Table 1 Radix Et Caulis Acanthopanacis Senticosi polysaccharide respectively organizes hepatic pathology degree of injury hierarchical table to Immune liver injury
4.2 serological index ALT, AST, IL-2, IL-4, IFN-γ horizontal detection
Get mice serum, by test kit, detection is described, the results are shown in Table 2, 3 and Fig. 9, 10, by table 2, 3 and Fig. 9, 10 is known, model group compares ALT with blank group, AST level raises, there is pole significant difference (P<0.01), immunologic liver injury modeling success is described, positive drug (bifendate) group, PDTC inhibitor matched group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group ALT compared with model group, AST, IL-2, IL-4, IFN-γ decreases, and have significant difference (P<0.05), in Radix Et Caulis Acanthopanacis Senticosi polysaccharide, though low dose group does not have significant difference compared with model group, but also adjust back to a certain extent.Illustrate that Radix Et Caulis Acanthopanacis Senticosi polysaccharide ASPS has the effect of the liver protecting and ALT lowering.
Table 2 Radix Et Caulis Acanthopanacis Senticosi polysaccharide on the impact of Immune liver injury serum transaminase (
n=10)
#,
*represent respectively with blank group, model group compares, p<0.05;
##,
*expression is compared with blank and model group, p<0.01
Table 3 Radix Et Caulis Acanthopanacis Senticosi polysaccharide on the impact of Immune liver injury serum IL-2, IL-4, IFN-γ (
n=10)
#,
*represent respectively with blank group, model group compares, p<0.05;
##,
*expression is compared with blank and model group, p<0.01
In 4.3 liver tissue homogenates, SOD, MDA, GSH-Px, NO, IL-1 β, TNF-alpha levels detects
Get liver tissue homogenate's liquid, adopt ELISA method to detect the content of NO, IL-1 β and TNF-α; Thiobarbituricacidα-colorimetric determination malonaldehyde (MDA) content; It is active that xanthine oxidase measures superoxide dismutase (SOD); Measure glutathion peroxidase (GSH-Px) vigor.The results are shown in Table 4,5 and Figure 11,12, by table 4 and the known model group of Figure 11, the GSH-PX compared in liver tissue homogenate reduces with blank group, and there is pole significant difference (P<0.01), positive drug (bifendate) group, PDTC inhibitor matched group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group numerical value compared with model group have significance to raise (P<0.01), and in Radix Et Caulis Acanthopanacis Senticosi polysaccharide, dosage group also has significant difference compared with model group; Model group compares MDA with blank group and raises, and there is pole significant difference (P<0.01), positive drug (bifendate) group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group numerical value compared with model group have significance to decline (P<0.01), and PDTC inhibitor matched group also has significance to decline compared with model group; Model group compares SOD with blank group and reduces, and there is pole significant difference (P<0.01), positive drug (bifendate) group, PDTC inhibitor matched group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group numerical value compared with model group have significance to raise (P<0.05), and in Radix Et Caulis Acanthopanacis Senticosi polysaccharide, low dose group also decreases compared with model group.From table 5 and Figure 12, model group compares the IL-1 β in liver tissue homogenate with blank group, NO, TNF-α has significance to raise (P<0.01), positive drug group, PDTC inhibitor matched group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group is compared with model group, IL-1 β numerical value has significance to adjust back reduction (P<0.05), NO and TNF-α all has significance to reduce (P<0.01) in positive drug group and Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group, the all variant readjustment (P<0.05) of both inhibitor groups.
Table 4 ASPS in Immune liver injury murine liver tissue GSH-Px, MDA, SOD impact (
n=10)
#,
*represent respectively with blank group, model group compares, p<0.05;
##,
*expression is compared with blank and model group, p<0.01
Table 5 ASPS in Immune liver injury murine liver tissue IL-1 β, NO, TNF-α affect (
n=10)
#,
*represent respectively with blank group, model group compares, p<0.05;
##,
*expression is compared with blank and model group, p<0.01
IL-2, IL-4, INF-γ horizontal detection in 4.4 homogenation of spleen tissue
Getting spleen tissue homogenate adopts ELISA method to detect the value of IL-2, IL-4 and INF-γ in mouse boosting tissue.All adopt double-antibody sandwich ABC-ELISA method.After microplate reader reading, formulate standard curve, be finally converted into the content of IL-2, IL-4 and INF-γ according to the standard curve formulated.Result is as shown in table 6 and Figure 13, from table 6 and Figure 13, IL-2, IL-4, INF-γ compared in homogenation of spleen tissue has significance to reduce (P<0.01) to model group with blank group, positive drug group, inhibitor matched group, Radix Et Caulis Acanthopanacis Senticosi polysaccharide high dose group are compared with model group, IL-2, IL-4, INF-γ numerical value has significance to reduce (P<0.05), the most obvious with IL-2 and IFN-γ.Radix Et Caulis Acanthopanacis Senticosi polysaccharide low dose group IL-2, IL-4, INF-γ numerical value also have reduction trend (P>0.05),
Radix Et Caulis Acanthopanacis Senticosi polysaccharide ASPS has certain prevention and protective effect to immunologic liver injury as seen from the experiment; polysaccharide is not only embodied directly in the liver protecting and ALT lowering aspect to the protective effect of hepatic injury; the activity of SOD and GSH-Px can also be improved from the ability improving hepatocellular anti-oxidative damage by suppressing MDA; its mechanism may remove body free radical with him, and enhancing body anti peroxidation of lipid ability is relevant with the expression of adhesion factor with reduction inflammatory cytokine.
Claims (3)
1. the novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide, is characterized in that the medicine of Radix Et Caulis Acanthopanacis Senticosi polysaccharide for the preparation for the treatment of immunologic liver injury.
2. the novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 1, is characterized in that the dosage of described medicine is 15.93mg/kg/d.
3. the novelty teabag of Radix Et Caulis Acanthopanacis Senticosi polysaccharide according to claim 1, is characterized in that described pharmaceutical administration is oral.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102138956A (en) * | 2011-03-31 | 2011-08-03 | 吉林延边朝药药业有限公司 | Health-care product having liver-protecting function and production process |
| CN104328030A (en) * | 2014-10-30 | 2015-02-04 | 国家开发投资公司 | Surface growth type culture plate having sandwich structure, surface growing culture system and surface growing culture method |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102138956A (en) * | 2011-03-31 | 2011-08-03 | 吉林延边朝药药业有限公司 | Health-care product having liver-protecting function and production process |
| CN104328030A (en) * | 2014-10-30 | 2015-02-04 | 国家开发投资公司 | Surface growth type culture plate having sandwich structure, surface growing culture system and surface growing culture method |
Non-Patent Citations (2)
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| 杨书良: "刺五加多糖的研究现状及进展", 《黑龙江医药》 * |
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