CN108685979A - The application of cat's whisker leaf extract - Google Patents

The application of cat's whisker leaf extract Download PDF

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Publication number
CN108685979A
CN108685979A CN201810934307.7A CN201810934307A CN108685979A CN 108685979 A CN108685979 A CN 108685979A CN 201810934307 A CN201810934307 A CN 201810934307A CN 108685979 A CN108685979 A CN 108685979A
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kidney
cat
whisker
group
mouse
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CN108685979B (en
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许丛丛
刘德坤
蔡旋
张璐
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Qufu Normal University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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  • Animal Behavior & Ethology (AREA)
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  • Urology & Nephrology (AREA)
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Abstract

The invention discloses application of the cat's whisker leaf extract in preparing the drug for treating injury of kidney.It is confirmed by testing, OS has regulating and controlling effect to the murine chronic kidney injury of Induced by High Fat Diet, the OS of various dose can reduce the mouse kidney damage of high fat diet induction to some extent, the most apparent with high dose OS (500mg/kg) effects, this can preferably reduce hyperlipidemia and oxidative stress in high fat diet Mice Body with it much relations.As it can be seen that cat's whisker leaf extract will have a wide range of applications in preparing the drug for treating injury of kidney.

Description

The application of cat's whisker leaf extract
Technical field
The invention belongs to biomedicine technical fields, and in particular to the application of cat's whisker leaf extract (OS).
Background technology
Mao Xu Cao (OSrodendranthus spicatus (Thunb.) C.Y.Wu) also known as kidney tea, cat's whisker are public, have " all over the world First is fresh " title, rich in flavonoids, phenols, terpene, lignanoids, chromogen alkenes, alkyl glycosides, steroid saponin, anthraquinone The a variety of active ingredients such as class, amino acid, organic acid, vitamin, minerals.The researchs such as Yam find Mao Xu Cao water and methanol extraction Object can preferably remove DPPH free radicals, inhibit Fe3+The lipid peroxidation of ion induction, hence it is evident that improve CCl4The rat of induction Hepatic injury.Sriplang etc. has found that Mao Xu Cao water extract can significantly reduce content of triglyceride in diabetes rat blood plasma, And significantly improve high-density lipoprotein cholesterol content.Liu Xuhang etc. is established big with Cationic bovine serum albumin (C-BSA) Mouse Chronic Nephritis Model, find Mao Xu Cao can also preferably reduce the Urine proteins of C-BSA Nephritis Model rats, serum creatinine, Urea nitrogen levels improve the renal function of rat.In addition, transforming growth factor-β1(TGF-β1) may participate in outside adjusting messangial cell The secretion of matrix causes glomerulus structure and function abnormal, promotes glomerulosclerosis.The researchs such as woods Weiyuan find Mao Xu Cao pair TGF-β1Expression have obvious inhibiting effect, fibronectin and collagen iv can be reduced in messangial cell epimatrix Deposition, delays glomerulosclerosis.Long Heming etc. has found that Mao Xu Cao general flavone also has the function of inhibiting renal carcinoma cells growth, this It may achieve the effect that inhibit growth of cancer cells related with the regulation and control by cell cycle and Apoptosis.As it can be seen that Mao Xu Cao exists Anti-oxidant, reducing blood lipid and improvement chronic nephritis etc. are with obvious effects.
In addition, it has now been found that high lipid diet causes abnormalities of sugar/lipid metabolism, induction hyperlipidemia, is to lead to Chronic Renal Impairment An important factor for.Kidney oxidative stress is likely to also play an important role wherein.However, Mao Xu Cao causes high fat diet The regulating effect of hyperlipidemia, oxidative stress and kidney injury, has not been reported.
Invention content
Goal of the invention:For the deficiencies in the prior art, the object of the present invention is to provide a kind of extractions of cat's whisker blade of grass Application of the object in preparing the drug for treating injury of kidney, satisfies the use demand.
Technical solution:In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:
Application of the cat's whisker leaf extract in preparing the drug for treating injury of kidney.
The injury of kidney is the injury of kidney generated by high fat diet.
The cat's whisker leaf extract is prepared by following methods:
Cat's whisker blade of grass 500g is taken, 50% ethyl alcohol soaking at room temperature of 25L is added for 24 hours, is stirred when impregnating, at 280W ultrasonic waves Manage 30min;Leaching liquor is filtered using middling speed quantitative filter paper, is freeze-dried after rotary evaporation concentration, extract is protected in drier It deposits.
20~50 μm of the aperture of the middling speed quantitative filter paper.
Advantageous effect:Compared with prior art, OS of the invention is to the murine chronic kidney injury of Induced by High Fat Diet With regulating and controlling effect, the OS of various dose can reduce the mouse kidney damage of high fat diet induction to some extent, with high dose OS (500mg/kg) effects are the most apparent, this can preferably reduce hyperlipidemia and oxidative stress in high fat diet Mice Body with it Level has much relations.It is widely answered as it can be seen that cat's whisker leaf extract will have in preparing the drug for treating injury of kidney With.
Description of the drawings
Fig. 1 is influence (n=6) figure of cat's whisker leaf extract to mouse kidney index;
Fig. 2 is Mao Xu Cao to mice serum triglycerides (TG), low-density protein cholesterol (LDL-C), high density egg Influence (n=6) figure of white matter cholesterol (HDL-C), total cholesterol (TC) content;
Fig. 3 is cat's whisker leaf extract on kidney of mouse GSH-Px (A) and active influence (n=6) results of SOD (B) Figure;
Fig. 4 is that cat's whisker leaf extract is homogenized H to kidney of mouse2O2(A), NO (B) and influence (n=of MDA (C) content 6) result figure;
Fig. 5 is influence (× 400) result figure of cat's whisker leaf extract to kidney of mouse structure;In figure, A:Control group; B:Model group;C:High dose group;D:Middle dose group;E:Low dose group.
Specific implementation mode
With reference to specific embodiment, the present invention is described further.
Main material used in following embodiment is as follows:
Experimental animal:Using 84 15~20g male C57BL/6 mouse, it is purchased from the limited duty of Shanghai Si Laike experimental animals Ren companies (animal productiong licensing number:SCXK (Shanghai) 2017-0005).
Animal feed:Common standard feed (D12450B), high lipid food (D12492) are by Research Diet Inc., New Brunswick, USA are provided.
Low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), total courage Sterol (TC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malonaldehyde (MDA), hydrogen peroxide (H2O2), nitric oxide (NO) kit and haematoxylin, eosin stain be purchased from and build up bio-engineering corporation in Nanjing;Penta 2 Aldehyde is purchased from Sinopharm Chemical Reagent Co., Ltd..
Embodiment 1
1, the extracting method of cat's whisker leaf extract (OS):Cat's whisker blade of grass 500g is taken, the leaching of 50% ethyl alcohol room temperatures of 25L is added Bubble for 24 hours, stirs, 280W ultrasonications 30min when impregnating.Leaching liquor uses middling speed quantitative filter paper (20~50 μm of aperture) mistake Filter is freeze-dried after rotary evaporation concentration, weighs powder weight, and it is 9.89% to calculate yield, and extract preserves in drier.
2, mice group and modeling:By 60 15~20g male C57BL/6 mouse, 5 groups are randomly divided into, is respectively compareed Group, model group, high dose (HOS) group, middle dosage (MOS) group and low dosage (LOS) group, every group 12.
Mouse is fitted in clean feeding environment (illumination/interlunation is 12h/12h, and temperature control is 23 ± 2 DEG C) Answering property is fed 1 week, and normal diet is gradually changed to experiment feed in the laundering period.Experimental period is 8 weeks, in experimental period, control group Mouse feeding common standard feed (D12450B), remaining group mouse feeding high lipid food (D12492).HOS groups, MOS groups and LOS It is respectively 500mg/kg, 100mg/kg, 50mg/kg that group mouse, which is fed for OS dosage, and control group is then fed for equivalent with model group mouse PBS (0.027%NaH2PO4, 0.142%Na2HPO4, 0.8%NaCl, 0.02%KCl).Mouse is freely eaten in experimental period, supplies To enough aqua sterilisas.It is deprived of food but not water after last gavage for 24 hours, plucks eyeball and take blood to be measured.After mouse takes blood, at cervical dislocation Extremely, it dissects, quickly removes kidney, observe form and weigh, a part of renal tissue is placed in 4% glutaraldehyde and fixes, and is seen for HE It examines;Remainder tissue is placed in Rapid-Freezing Method in liquid nitrogen, and -80 DEG C of preservations are to be measured.
3, mouse kidney index:Renal index (%)=kidney weight (g)/weight (g) × 100%.
4, the measurement of mice serum Biochemical Indexes:Plucking eyeball takes blood, whole blood to be stored at room temperature 30min, 4 DEG C of items after blood coagulation 3000rpm centrifuges 10min under part, draws serum, and LDL-C, HDL-C, TC and TG content are detected by kit specification step.
5, the measurement of kidney of mouse homogenate Biochemical Indexes:Mouse kidney tissue accurately is weighed, by 1: 9 (m:V) ratio 0.9% physiological saline of 9 times of volumes is added in example, 10% tissue homogenate is made, 4000r/min is centrifuged under the conditions of 4 DEG C 10min takes supernatant, and GSH-Px and SOD activity and H are measured by kit specification step2O2, NO and MDA contents.
6, kidney of mouse HE is dyed:With reference to Wang Xiangli[12]Report method takes about 1cm3Renal tissue, 4% glutaraldehyde are molten Fixed in liquid routine paraffin wax embeds for 24 hours, and 5 μ m-thicks slice is made.Then, it is dyed according to Hematoxylin-eosin (HE) sequence, then With neutral gum mounting.Finally, nephridial tissue structure is observed under an optical microscope.
7, data processing
Data analysis is carried out using SPSS18.0 statistical softwares, experimental result is indicated with x ± S, and indicates aobvious with P < 0.05 Write otherness.
As shown in Figure 1, model group mouse kidney index is significantly higher than control group (P < 0.05).Compared with model group, OS high The renal index of dosage group, middle dose group and low dose group mouse is remarkably decreased (P < 0.05).As can be seen that OS is to subtracting The mouse kidney index elevating effect of slow high fat diet induction is apparent.
As shown in Fig. 2, compared with the control group, TG, LDL-C and TC content significantly increase (P < in model group mice serum 0.05), and HDL-C contents be in notable downward trend (P < 0.05).These results illustrate that high fat diet leads to blood fat in Mice Body There is hyperlipidemia in metabolic disorder.
Compared with model group, TG, LDL-C and TC content are equal in OS high dose groups, middle dose group, low dose group mice serum There is different degrees of reduction.Each group effect size compares:Di Jiliangzu <Zhong Jiliangzu <High dose group.With OS high dose group mouse blood TG, LDL-C and TC content reduce the most apparent in clear, are in significant difference (P < 0.05).Wherein, TG, LDL-C and TC content Reduction amplitude respectively reaches 13.45%, 29.68% and 16.87%.Though middle dose group and low dose group have reduction trend, not Significantly (P > 0.05).In each OS reparations group, significantly (P < are increased with HDL-C contents in high dose group mice serum 0.05), other two groups of raising unobvious (P > 0.05).As can be seen that OS can reduce height in the Mice Body of high fat diet induction Blood lipid level, and have certain dosage correlation, high dose group effect is maximum.
As shown in figure 3, GSH-Px and SOD activity reduces respectively compared to control group in model group kidney of mouse 53.64% and 33.23% (P < 0.05).GSH-Px and SOD activity is all remarkably higher than model in each OS reparations group kidney of mouse Group (P < 0.05).GSH-Px and SOD activity is respectively model in high dose group, middle dose group and low dose group kidney of mouse Group 1.53 times and 1.23 times, 1.45 times and 1.19 times, 1.46 times with 1.20 times.Each OS reparations group acts on size without significant difference (P > 0.05).As can be seen that OS has apparent effect to GSH-Px and SOD antioxidant systems in enhancing high fat diet kidney of mouse Fruit.
Fig. 4 shows, compared with the control group, H in model group kidney of mouse2O2, NO and MDA contents significantly increase (P < 0.05), illustrate that mouse kidney tissue is in oxidative stress status.
Compared with model group, H in high dose group, middle dose group and low dose group kidney of mouse2O2Content significantly drops Low (P < 0.05), reduction amplitude are up to 18.54%, and effect size there is not significant difference (P > 0.05) between each OS reparations group.Phase Compared with model group, NO contents reduce 32.23% (P < 0.05), middle dose group and low dose group in high dose group mouse kidney NO changes of contents is not notable (P > 0.05).MDA contents are equal compared to model group in high dose group and middle dose group kidney of mouse It significantly reduces (P < 0.05), reduces 37.32% and 12.39% respectively.High dose group effect degree is significantly higher than middle dose group (P < 0.05).MDA contents are compared to model group variation unobvious (P > 0.05) in low dose group kidney of mouse.It can see Go out, OS has positive effect to reducing high fat diet inducing mouse kidney oxidative stress, and certain dosage correlation, high agent is presented It is the most apparent to measure OS regulating effects.
In 400 times of microscopic observation mouse kidney tissue HE stained slices as it can be seen that control group mesonephric vesicle surrounds glomerulus, Malpighian corpuscle morphologic features are normal, renal tubule form rule, (Fig. 5 A) clear in structure;Model group renal tissue section is pale, portion Divide glomerulus loose, cell number increases, and proliferation of mesangial cells, extracellular matrix is broadening, and some changes (perpendicular arrow institute in lobulated Show).Glomerulus cyst membrane is irregular, and structure is imperfect, and blister cavities increases (shown in horizontal arrow).Renal tubule structure is unintelligible and has apparent Vacuolar degeneration is damaged, the visible renal tubule atrophy (shown in asterisk) (Fig. 5 B) having;OS high dose groups, middle dose group and low dose group In, mesangial cell is inhibited with matrix secreted, and capsula glomeruli film improves in various degree with tubular degeneration damage (Fig. 5 C-E);High dose group improvement is better than middle dose group and low dose group.As can be seen that OS is to high fat diet inducing mouse Kidney structure damage, which has, obviously slows down effect, and high dose OS improvements are the most apparent.
As it can be seen that OS has regulating and controlling effect to the murine chronic kidney injury of Induced by High Fat Diet, the OS of various dose can The mouse kidney damage of high fat diet induction is reduced to some extent.It is the most apparent with high dose OS (500mg/kg) effects, this Can preferably reduce hyperlipidemia and oxidative stress in high fat diet Mice Body with it has much relations.

Claims (4)

1. application of the cat's whisker leaf extract in preparing the drug for treating injury of kidney.
2. application according to claim 1, which is characterized in that the injury of kidney is to be damaged by the kidney that high fat diet generates Wound.
3. application according to claim 1, which is characterized in that the cat's whisker leaf extract is prepared by following methods:
Cat's whisker blade of grass 500g is taken, 50% ethyl alcohol soaking at room temperature of 25L is added for 24 hours, is stirred when impregnating, 280W ultrasonications 30min;Leaching liquor is filtered using middling speed quantitative filter paper, is freeze-dried after rotary evaporation concentration, extract preserves in drier.
4. application according to claim 3, which is characterized in that 20~50 μm of the aperture of the middling speed quantitative filter paper.
CN201810934307.7A 2018-08-16 2018-08-16 Application of radix ranunculi ternati leaf extract Active CN108685979B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114191532A (en) * 2021-11-02 2022-03-18 桂林理工大学 Uric acid reducing composition, preparation method and application thereof
CN116036150A (en) * 2022-12-30 2023-05-02 深圳市中医院 Application of radix ranunculi ternati in preparation of medicines for treating kidney fibrosis

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114191532A (en) * 2021-11-02 2022-03-18 桂林理工大学 Uric acid reducing composition, preparation method and application thereof
CN114191532B (en) * 2021-11-02 2022-11-11 桂林理工大学 Uric acid reducing composition, preparation method and application thereof
CN116036150A (en) * 2022-12-30 2023-05-02 深圳市中医院 Application of radix ranunculi ternati in preparation of medicines for treating kidney fibrosis

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