CN116687914B - Medicine for treating non-alcoholic fatty liver - Google Patents
Medicine for treating non-alcoholic fatty liver Download PDFInfo
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- CN116687914B CN116687914B CN202310981801.XA CN202310981801A CN116687914B CN 116687914 B CN116687914 B CN 116687914B CN 202310981801 A CN202310981801 A CN 202310981801A CN 116687914 B CN116687914 B CN 116687914B
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- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 title claims abstract description 43
- 239000003814 drug Substances 0.000 title claims abstract description 28
- 229940079593 drug Drugs 0.000 title description 5
- 150000002596 lactones Chemical class 0.000 claims abstract description 35
- 241000132446 Inula Species 0.000 claims description 58
- 235000021588 free fatty acids Nutrition 0.000 claims description 21
- 210000005229 liver cell Anatomy 0.000 abstract description 26
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 abstract description 15
- 230000006372 lipid accumulation Effects 0.000 abstract description 8
- 208000004930 Fatty Liver Diseases 0.000 abstract description 6
- 206010019708 Hepatic steatosis Diseases 0.000 abstract description 6
- 208000010706 fatty liver disease Diseases 0.000 abstract description 6
- 231100000240 steatosis hepatitis Toxicity 0.000 abstract description 6
- 230000032683 aging Effects 0.000 abstract description 5
- 241000253121 Inula britannica Species 0.000 abstract description 2
- 241000533042 Inula grandiflora Species 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 30
- 210000004027 cell Anatomy 0.000 description 25
- 230000000694 effects Effects 0.000 description 13
- 239000000203 mixture Substances 0.000 description 12
- 125000000686 lactone group Chemical group 0.000 description 9
- 210000003494 hepatocyte Anatomy 0.000 description 7
- 238000013232 NAFLD rodent model Methods 0.000 description 5
- 150000002632 lipids Chemical group 0.000 description 5
- 238000011160 research Methods 0.000 description 5
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- 239000000243 solution Substances 0.000 description 4
- 102000005936 beta-Galactosidase Human genes 0.000 description 3
- 108010005774 beta-Galactosidase Proteins 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
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- JXEGMONJOSAULB-UHFFFAOYSA-N (9-acetyloxy-8-hydroxy-5,8a-dimethyl-1-methylidene-2-oxo-4,5,5a,6,7,8,9,9a-octahydro-3ah-azuleno[6,5-b]furan-6-yl) acetate Chemical compound CC1CC2OC(=O)C(=C)C2C(OC(C)=O)C2(C)C(O)CC(OC(C)=O)C12 JXEGMONJOSAULB-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
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- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- NNDHDYDFEDRMGH-CAEIVAEBSA-N Anthranoyllycoctonine Chemical compound C([C@]12CN(C3[C@@]4(O)[C@]5(O)[C@H]6[C@@H](OC)[C@@H]([C@H](C5)OC)C[C@H]6[C@@]3([C@@H]1[C@@H]4OC)[C@@H](OC)CC2)CC)OC(=O)C1=CC=CC=C1N NNDHDYDFEDRMGH-CAEIVAEBSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010009208 Cirrhosis alcoholic Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- UXOXDDUEWZOAIW-UHFFFAOYSA-N Inuline Natural products CCN1CC2(CC(=O)Oc3ccccc3N)CCC(OC)C45C6CC7C(CC(O)(C6C7OC)C(O)(C(OC)C24)C15)OC UXOXDDUEWZOAIW-UHFFFAOYSA-N 0.000 description 1
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- JXEGMONJOSAULB-NKNCGKRASA-N [(3as,5r,5as,6r,8s,8as,9s,9ar)-9-acetyloxy-8-hydroxy-5,8a-dimethyl-1-methylidene-2-oxo-4,5,5a,6,7,8,9,9a-octahydro-3ah-azuleno[6,5-b]furan-6-yl] acetate Chemical group C[C@@H]1C[C@@H]2OC(=O)C(=C)[C@H]2[C@H](OC(C)=O)[C@]2(C)[C@@H](O)C[C@@H](OC(C)=O)[C@@H]12 JXEGMONJOSAULB-NKNCGKRASA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 208000010002 alcoholic liver cirrhosis Diseases 0.000 description 1
- VNRZCPPHNPEBFC-UHFFFAOYSA-N anthranoyllycoctonine Natural products CCN1CC2(COC(=O)c3ccccc3N)CCC(OC)C45C2C(OC)C(O)(C14)C6(O)CC(OC)C7CC5(O)C6C7OC VNRZCPPHNPEBFC-UHFFFAOYSA-N 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 206010053219 non-alcoholic steatohepatitis Diseases 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
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- 208000011580 syndromic disease Diseases 0.000 description 1
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- 230000009385 viral infection Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a medicine for treating non-alcoholic fatty liver, and belongs to the technical field of fatty liver treatment. The invention discovers that the inula grandiflora lactone can play a role in treating the non-alcoholic fatty liver by reducing the triglyceride content in liver cells, reducing the aging of the liver cells and reducing the lipid accumulation of the liver cells. Therefore, the preparation method can be used for preparing the medicine for treating the non-alcoholic fatty liver. The invention not only widens the application range of the inula britannica lactone, but also provides a novel safe and reliable medicament for treating the non-alcoholic fatty liver.
Description
Technical Field
The invention belongs to the technical field of fatty liver treatment, and particularly relates to a medicine for preventing and treating nonalcoholic fatty liver.
Background
Nonalcoholic fatty liver disease (NAFLD) refers to a pathological syndrome characterized by excessive deposition of lipid in liver cells, which is caused by removal of alcohol and other definite liver-damaging factors (such as drugs, viral infections or autoimmunity). According to the development process, non-alcoholic fatty liver disease can be classified into simple fatty liver, non-alcoholic steatohepatitis and cirrhosis. At present, the prevalence rate of the non-alcoholic fatty liver disease in the global scope is rapidly rising, and as the pathogenesis of the non-alcoholic fatty liver disease is complex and various, no specific medicine specially used for treating the non-alcoholic fatty liver disease is available at present, so the research on a treatment method and a treatment medicine for the non-alcoholic fatty liver disease is a hot spot of current research.
Inula flower lactone (Britanin), also known as inuline, is a plant extract from Inula flower of formula C 19 H 26 O 7 The CAS number is 33627-28-0. The existing research shows that the inula flowers lactone has low toxicity to normal cells and can produce effective anticancer effect. In addition, the existing research shows that the inula britannica lactone also has the effect of improving brainIschemic reperfusion injury. However, the effect and mechanism of the inula variegata lactone in the fatty liver treatment are not related to the related content of the inula variegata lactone in the fatty liver treatment at present, so the invention researches the effect and mechanism of the inula variegata lactone in the fatty liver treatment.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a new application of inula major flowers lactone and a medicine capable of effectively treating non-alcoholic fatty liver.
The first object of the invention is to provide the application of inula major in preparing the medicine for treating the non-alcoholic fatty liver.
Preferably, the medicament acts to treat non-alcoholic fatty liver disease by reducing triglyceride levels in hepatocytes.
Preferably, the medicament acts to treat non-alcoholic fatty liver disease by reducing aging of hepatocytes.
Preferably, the medicament acts to treat non-alcoholic fatty liver disease by reducing lipid accumulation in liver cells.
Preferably, the non-alcoholic fatty liver disease is a non-alcoholic fatty liver disease caused by free fatty acids.
Preferably, the concentration of inula major flowers is 31.25nM-1000nM.
Preferably, the optimal concentration of inula major flower lactone is 125nM.
A second object of the present invention is to provide a medicament for treating non-alcoholic fatty liver disease, characterized in that the active ingredient of the medicament is inula major-flowers lactone.
Preferably, the medicament acts to treat non-alcoholic fatty liver disease by reducing triglyceride levels in hepatocytes;
the medicine can be used for treating nonalcoholic fatty liver by reducing liver cell aging;
the medicament plays a role in treating non-alcoholic fatty liver by reducing lipid accumulation of liver cells.
Preferably, the non-alcoholic fatty liver disease is a non-alcoholic fatty liver disease caused by free fatty acids.
Preferably, the concentration of inula major flowers is 31.25nM-1000nM.
Preferably, the optimal concentration of inula major flower lactone is 125nM.
A third object of the present invention is to provide the use of inula major in the manufacture of a medicament for reducing the increase in triglyceride content in hepatocytes caused by free fatty acids.
Preferably, the concentration of inula major flowers is 31.25nM-1000nM.
Preferably, the optimal concentration of inula major flower lactone is 125nM.
The fourth object of the invention is to provide the application of inula major in preparing the medicine for reducing the liver cell aging caused by free fatty acid.
Preferably, the concentration of inula major flowers is 31.25nM-1000nM.
Preferably, the optimal concentration of inula major flower lactone is 125nM.
A fifth object of the present invention is to provide the use of inula major in the manufacture of a medicament for reducing the accumulation of hepatocyte lipids caused by free fatty acids.
Preferably, the concentration of inula major flowers is 31.25nM-1000nM.
Preferably, the optimal concentration of inula major flower lactone is 125nM.
The invention has the following beneficial effects:
the invention provides a new application of inula flower lactone in preparing a non-alcoholic fatty liver disease treatment drug, and discovers that the inula flower lactone can play a role in treating the non-alcoholic fatty liver disease by reducing triglyceride content in liver cells, reducing aging of the liver cells and reducing lipid accumulation of the liver cells. Therefore, the invention not only widens the application range of the inula major flowers lactone, but also provides a new safe and reliable medicament for treating the non-alcoholic fatty liver.
Drawings
FIG. 1 shows the cytotoxicity test results of inula flower lactone on liver cell LO-2;
FIG. 2 shows the results of the detection of the effect of inula major on TG content in a non-alcoholic fatty liver cell model;
FIG. 3 is a graph showing the results of the detection of the effect of inula major on cell senescence in a non-alcoholic fatty liver cell model;
FIG. 4 shows the results of the detection of the effect of inula major on lipid accumulation in a non-alcoholic fatty liver cell model.
Detailed Description
Example 1: selecting inula flowers lactone at a concentration gradient: 1000nM,500nM,250nM,125nM,62.5nM,31.25nM,0 nM:
(1) The LO-2 cells cultured after resuscitation were digested and counted, and the concentration of the cell suspension was adjusted to 5X 10 4 Individual/ml;
(2) Adding 100ul of cell suspension into a cell culture hole for culturing for 24 hours, discarding the culture medium one by one, and repeatedly flushing with PBS buffer solution for 2 times;
(3) Diluting inula flower lactone (purchased from Shanghai source leaf biotechnology Co., ltd.) to a desired concentration with 1640 complete medium, and adding 200ul of the corresponding gradient of the drug-containing medium to the cell culture plate;
(4) The cell culture plates were placed in an incubator for 24 hours and stained for CCK-8, OD was measured and cell activity was calculated for each group.
Firstly, the invention detects whether the inula variegata lactone has toxicity to the liver cell LO-2, and the result of figure 1 shows that the inula variegata lactone has no obvious effect on the cell activity of the liver cell LO-2 in the concentration range of 0-1000nM, which indicates that the inula variegata lactone has no toxicity to the liver cell and can be used for preparing medicaments.
Example 2: (1) 0.0609g of oleic acid and 0.0278g of palmitic acid are weighed by using an analytical balance, sequentially added into 15ml of 25% bovine serum albumin PBS solution, heated at 70 ℃, and then put into ultrasound until the mixture is completely dissolved;
(2) Diluting the prepared 20mM mother solution into 1mM free fatty acid mixed solution, and filtering and sterilizing by using a 0.22um filter for later use;
(3) LO-2 cells were inoculated evenly into petri dishes and after 24h cell attachment were treated according to the following groupings:
normal group: replacing 1640 complete medium;
NAFLD model group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture;
low dose of inula flowers lactone group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture +31.25nM inula major;
medium dose of inula flowers lactone group: the medium was changed to 1640 complete medium containing 1mM free fatty acid mixture +125nM inula major lactone;
high dose inula flowers lactone group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture +250nM inula major;
(4) After 24h of treatment, the culture solution was aspirated, washed 2 times with PBS, 1ml of PBS was added and the cells were scraped off with a cell scraper, collected into a centrifuge tube with a pipette, centrifuged at 1000rpm at 4℃for 5min;
(5) The supernatant was discarded, 200ul of 2% Triton X-100 lysate was added, and the mixture was lysed on ice for 40min, and the intracellular Triglyceride (TG) content was detected with reference to the TG kit.
The increase in triglyceride was the main cause of non-alcoholic fatty liver, so in example 2, the present invention examined the effect on TG content in a non-alcoholic fatty liver cell model, and the results are shown in fig. 2.
As can be seen from fig. 2, the TG content of the NAFLD model group was significantly increased compared with the normal group, while the TG content of the low, medium and high dose inula flowers lactone group was somewhat decreased compared with the NAFLD model group, and the difference was statistically significant, indicating that the treatment of the nonalcoholic fatty liver cell model with inula flowers lactone can effectively inhibit the TG content in the cells, and the concentration of about 125nM was optimal.
Example 3: (1) LO-2 cells were inoculated evenly into petri dishes and after 24h cell attachment were treated according to the following groupings:
normal group: replacing 1640 complete medium;
NAFLD model group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture;
low dose of inula flowers lactone group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture +31.25nM inula major;
medium dose of inula flowers lactone group: the medium was changed to 1640 complete medium containing 1mM free fatty acid mixture +125nM inula major lactone;
high dose inula flowers lactone group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture +250nM inula major;
(2) After 24h of treatment, the culture medium is sucked off, PBS is added to wash the cells for 2 times, and the cell fixative is added to fix the cells for 15min;
(3) Removing the fixing solution, adding PBS to wash the cells for 2 times, adding beta-galactosidase staining solution, sealing by using a preservative film, and then placing the cells at 37 ℃ for 12 hours of incubation;
(4) The staining solution was removed, positive cells were counted and the positive rate was calculated.
The aging of liver cells is also an important cause of nonalcoholic fatty liver disease caused by free fatty acid, so the effect of inula flower lactone on cell aging in a nonalcoholic fatty liver cell model is detected, and the result is shown in figure 3.
As can be seen from fig. 3, the proportion of β -galactosidase positive cells of the model group was significantly increased compared to the control group, while the proportion of β -galactosidase positive cells of the low, medium and high-dose strophaneroctone group was significantly decreased compared to the model group, and the effect was optimized at the medium dose as in example 2. The Inula flower lactone can effectively inhibit hepatic cell aging caused by free fatty acid.
Example 4: (1) LO-2 cells were inoculated evenly into petri dishes and after 24h cell attachment were treated according to the following groupings:
normal group: replacing 1640 complete medium;
NAFLD model group: the medium was replaced with 1640 complete medium containing 1mM free fatty acid mixture;
inula flowers lactone group: the medium was changed to 1640 complete medium containing 1mM free fatty acid mixture +125nM inula major lactone;
(2) After 24h of treatment, the culture solution is sucked away, PBS is used for cleaning for 2 times, and 4% paraformaldehyde is added to fix the cells for 15min;
(3) Sucking away paraformaldehyde, washing cells with PBS for 2 times, adding oil red O working solution, and dyeing for 10min in dark place;
(4) Rinsing with 60% isopropanol for several seconds to remove excess dye;
(5) Adding hematoxylin staining solution for 1.5min, rinsing with double distilled water for 3 times, and photographing under an inverted microscope.
Hepatocyte lipid accumulation is a major manifestation of non-alcoholic fatty liver, so the invention detects the effect of inula flowers lactone on lipid accumulation in non-alcoholic fatty liver cell models. As can be seen from fig. 4, the number of lipid droplets in the model group was significantly increased compared to the normal group, while the number of lipid droplets in the inula flower lactone group was significantly decreased compared to the model group, indicating that inula flower lactone can effectively reduce lipid accumulation in cells.
In the embodiment, the inula flower lactone can effectively reduce the TG content increase in the liver cells caused by the free fatty acid, and the aging of the liver cells and the accumulation of the lipid in the liver cells, thereby effectively treating the nonalcoholic fatty liver caused by the free fatty acid.
Claims (1)
1. The application of inula flower lactone in preparing medicine for treating non-alcoholic fatty liver caused by free fatty acid is characterized in that,
the concentration of the inula flowers lactone in the medicine is 31.25nM-250nM.
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| CN202310981801.XA CN116687914B (en) | 2023-08-07 | 2023-08-07 | Medicine for treating non-alcoholic fatty liver |
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| CN202310981801.XA CN116687914B (en) | 2023-08-07 | 2023-08-07 | Medicine for treating non-alcoholic fatty liver |
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| CN116687914B true CN116687914B (en) | 2024-01-16 |
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| CN102369202A (en) * | 2008-11-13 | 2012-03-07 | 链接医药公司 | Azaquinolinone derivatives and uses thereof |
| WO2013152313A1 (en) * | 2012-04-05 | 2013-10-10 | The Regents Of The University Of California | Compositions and methods for treating cancer and diseases and conditions responsive to growth factor inhibition |
| KR20140112636A (en) * | 2013-03-12 | 2014-09-24 | 영남대학교 산학협력단 | Composition for inhibiting cellular senescence comprising extracts of Inula japonica Thunberg or britanin isolated from the same |
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