CN108498566A - A kind of application of Astragalus Root P.E in treatment cadmium causes diabetic nephropathy - Google Patents
A kind of application of Astragalus Root P.E in treatment cadmium causes diabetic nephropathy Download PDFInfo
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Abstract
The present invention provides a kind of application of Astragalus Root P.E in treatment cadmium causes diabetic nephropathy.Joint intraperitoneal injection caddy induction is fed using high sugar high in fat, structure cadmium causes diabetic nephropathy mouse model.Cause the protective effect of pathogenesis of diabetic nephropathy, feature and Radix Astragali to carry out experimental study cadmium from oxidative stress, correlative protein expression level etc. are many-sided, for treatment by long-term cadmium expose and high-carbonhydrate diet high in fat caused by diabetic nephropathy provide new approach.
Description
Technical field
The invention belongs to area of pharmacology, are related to the activity of Radix Astragali and its extract for treating diabetic nephrosis, more particularly to
The treatment of diabetic nephropathy caused by long-term cadmium exposure and high-carbonhydrate diet high in fat.
Background technology
Diabetes (diabetes mellitus, DM) are global serious public health problems, and diabetes prevalence goes out
Now acutely increase trend.Diabetes harmfulness is mainly manifested on various chronic complicating diseases, is not only the lethal cause of diabetic
Residual main reason, also creates heavy financial burden.
Diabetic nephropathy (diabetic nephropathy, DN) is diabetes generalized capillary complication, is also led
T2DM patient's accumulation of one of the main reason for causing chronic kidney hypofunction, course of disease 10 years or more has 20% generation DN.In western countries, DN
The Etiological for having become end-stage renal disease (end-stage renal disease, ESRD), in the U.S., nearly 42% ESRD
Patient is caused by DM, and wherein T2DM is even more to have accounted for 90%, and in ESRD newly-increased every year, 50% is DN patient.And according to
The data of China urban statistics in 2002 shows that DN accounts for 15% of ESRD or so.DN early clinic symptoms are hidden, and find evening
And lesion is difficult to reverse, and still lacks effective therapy at present, only relies on hypoglycemic, decompression and adjusts the conventional therapies means such as fat.
Therefore, the current treatment status of DN allows of no optimist, and key is to early diagnose and prevent, once into the clinical proteinuria phase, kidney damage
It is harmful then be difficult to reverse.The pathogenesis of DN is more complex, completely not clear yet so far, is related to high sugared toxicity, disorders of lipid metabolism, kidney blood
The many aspects such as hydromechanics is abnormal, cell factor and growth factor, inflammatory factor and oxidative stress, and in any of the above mechanism
Oxidative stress is the Common Mechanism of DN morbidities.
Cadmium (Cd) is a kind of toxic heavy metal, is regarded as by noxious material and disease registration office joint institution most dangerous
One of environmental contaminants.Cd deprofessionaliztions exposure and the main source of poisoning are drinking water, food pollution and smoking.Cd once into
Entering in vivo gradually to accumulate, and long half time reaches decades.Acute and chronic Cd exposures, can act on different organs and group
It knits, such as liver, kidney, lung, intestines, central nervous system, ovary, testis, pancreas.Oxidative stress is the pass of body function cell damage
Key risk factors, are related to a variety of lysises and Cd causes one of the important mechanisms of organ toxicity, each organ that oxidative stress occurs
Mechanism major embodiment in the antioxidant defense mechanisms of cell.Long-term Cd exposures can cause body to generate lasting hyperglycemia,
Eventually lead to the generation of type-2 diabetes mellitus.
Radix Astragali (Astragalus Membranaceus) is the dry root of legume Inner Mongolia Astragalis or Astragalus membranacus, main
Want ingredient for astragalus saponin, astragalus polyose, 1- aminobutyric acids, trace element etc., it clinically can hypoglycemic, tune fat, reduction Urine proteins.
Radix Astragali first recorded in《This warp》, sweet in flavor, slightly warm in nature, for clinical conventional Chinese medicine, with tonifying Qi and lifting yang, hemostasis by invigorating QI, tonifying Qi row is stagnant, dispels
Stasis and resolving masses, invigorating qi for consolidating superficies, beneficial the moon of tonifying Qi, inducing diuresis for removing edema, it is warm in cold dispelling, the multiple pharmacological effects such as strengthen the body resistance to consolidate the constitution and slow down aging.
2010 editions《Chinese Pharmacopoeia》It is expressly recited：" Radix Astragali is used for qi deficiency edema, Heat Diabetes " etc., research confirms, Radix Astragali energy nephrectasia
Blood vessel increases renal blood flow, reduces albuminuria, improves glycolipid metabolism, protects renal blood vessels inner skin cell function, reduces blood platelet
The effects that degree of sticking, shows that Radix Astragali and its compound preparation can protect kidney, delays renal failure.Zhu Likun reports " Radix Astragali
To the adjustment effect of db/db mouse kidney inflammatory Cytokines Expressions ", the results showed that, Radix Astragali acts on without shadow the blood glucose of db/db mouse
It rings；To IL-10, IL-1 β, MCP-1 and the TNF-α of db/db mouse kidneys, these inflammatory factors have significant inhibition to make to Radix Astragali
With to the IL-4 of mouse spleen, IFN-γ, IL-6 and IL-10 effect unobvious；Radix Astragali can adjust db/db murine interleukins
With the quantity of monocyte.
But the active ingredient and mechanism of Astragalus in Treating DM, DN not yet illustrate completely, with modern molecular biology and now
For pharmaceutical technology, the mechanism of further investigation Astragalus in Treating DM and DN are conducive to find the new side for the treatment of diabetes, diabetic nephropathy
Method and approach are also beneficial to preferably develop and utilize Radix Astragali.
Invention content
The purpose of the present invention is to provide a kind of application of Astragalus Root P.E in treatment cadmium causes diabetic nephropathy.
In order to achieve the above objectives, present invention employs following technical schemes：
The present invention feeds joint intraperitoneal injection caddy induction using high sugar high in fat, constructs a kind of diabetic nephropathy mouse
Model.It is composed using Gas chromatographyMass spectrometry (GC-MS) detection blood serum metabolic profiling, in conjunction with statistics such as principal component analysis
Method analysis of control group, diabetic nephropathy model group and protection group metabolin are found and (the also referred to as cadmium cause of such diabetic nephropathy
Diabetic nephropathy) relevant biomarker occurs.From oxidative stress, metallic element level and correlative protein expression water
Flat various aspects analyse in depth the protective effect of such pathogenesis of diabetic nephropathy and Radix Astragali.
The result shows that diabetic nephropathy (the cadmium cause glycosuria that long-term cadmium exposure (such as smoking) and high-carbonhydrate diet high in fat are caused
Sick nephrosis) pathogenesis is complex, and there are multiple links and target spot, single target spot intervention effect is not good enough.Pass through astragalus extraction
Object intervenes diabetic nephropathy model, shows that Astragalus Root P.E causes diabetic nephropathy that there is significant protection to make cadmium
With can be applied in the drug for preventing such diabetic nephropathy.
Beneficial effects of the present invention are embodied in：
The present invention discloses Radix Astragali and causes diabetic nephropathy to have significant protection cadmium by the intervention effect of Astragalus Root P.E
Effect causes diabetic nephropathy to provide new approach for prevention cadmium.
Description of the drawings
Fig. 1 is separate groups of mice kidney transmission electron microscope ultra microstructure figure, wherein：A is K groups, and B is M groups, and C is S groups, D H
Group.
Fig. 2 is the blood glucose test results of mouse in different groups.
Fig. 3 is the blood TG testing results of mouse in different groups, wherein # is indicated compared with diabetic nephropathy group (i.e. M groups)
p<0.05, * * indicates the p compared with blank group (i.e. K groups)<0.01, * indicates the p compared with blank group (i.e. K groups)<0.05.
Fig. 4 is the blood TC testing results of mouse in different groups, wherein * * indicate the p compared with blank group (i.e. K groups)<
0.01, ## indicates the p compared with diabetic nephropathy group (i.e. M groups)<0.01, # indicates the p compared with diabetic nephropathy group (i.e. M groups)<
0.05。
Fig. 5 is the blood CRE testing results of mouse in different groups, wherein * * indicate the p compared with blank group (i.e. K groups)<
0.01, ## indicates the p compared with diabetic nephropathy group (i.e. M groups)<0.01, # indicates the p compared with diabetic nephropathy group (i.e. M groups)<
0.05。
Specific implementation mode
The present invention will be further described with reference to the accompanying drawings and examples.
One, animal prepares
Male mouse of kunming 70.
Two, animal model structure and packet transaction
1, it is grouped
At random to mice group, 7 groups are divided into, every group of 10 mouse.Respectively Normal group (K groups), cadmium induced module
Type group (M groups), Radix Astragali protection I groups (H groups), positive drug control I groups (Y groups), STZ control groups (S groups), Radix Astragali protect II groups
(H2 groups), positive drug control II groups (Y2 groups), referring to table 1.
2, drug is protected
1) preparation of Astragalus Root P.E
Milkvetch Root crushes, and crosses 40 mesh sieve, and it is 10g to take medicinal material amount, and pre-soaking 8 hours in water are flowed back using water as solvent
It extracts (about 100 DEG C), is extracted 2 times, first time 2h, second of 1h with l0 times of amount (mg/mL) of medicinal material, merging filtrate, rotation is steamed
After sending out instrument reduced pressure (about 40 DEG C), freeze drier freeze-drying pulverizes, obtains 1.9g dry extracts, paste-forming rate 19% uses this
Method prepares Radix Astragali dry extract until reaching the desired amount of Astragalus Root P.E.
2) positive control medicine
According to reported in literature, select for melbine of the DN with protective effect as a contrast.
3, model construction
1) animal selects：Select the bull kunming mice of 20g ± 2g.First by the mouse of experiment before modeling
Carry out adaptable fed 7 days, automatic feeding drinking-water.
2) adaptable fed is passed through one week to the male mouse of kunming of selection (M groups, H groups, Y groups, S groups, H2 groups and Y2 groups)
Afterwards, give high-sugar-fat-diet to feed 8 weeks, the component and quality proportioning of the high-sugar-fat-diet are as follows：Lard 18%, sucrose
20%, yolk 3% and normal diet 59%.
3) high-sugar-fat-diet feeds the processing of joint cadmium slow poisoning：During high-sugar-fat-diet is fed, at the same it is right
The 1mg/ CdCl of (kgd) are given in mouse (M groups, H groups and Y groups) intraperitoneal injection2.It feeds to after 6 weeks, to mouse fasting 12 hours
After measure its fasting blood-glucose, it is preliminary to distinguish whether model builds success using blood glucose >=11.1mmol/L as standard；Blood glucose value is reached
Mouse to standard further detects urine creatinine, urea nitrogen (BUN), Urinary microalbumin variation.Finally with blood glucose >=
11.1mmol/L and NAG >=21 ± 6U/L is diabetic nephropathy mouse modeling success indicator.
4) high-sugar-fat-diet feeds joint STZ processing：It is right at the end of high-sugar-fat-diet fed progress to the 6th week
50mg/ (kgd) STZ is given in positive control damage group (i.e. S groups), the injection in continuous three days of H2 groups and Y2 group mouse, waits for 72h blood glucose
Fasting blood sugar (fasting 12h) is measured after stabilization.It is preliminary to distinguish whether model is built into using blood glucose >=11.1mmol/L as standard
Work(；The mouse for reaching standard to blood glucose value further detects urine creatinine, urea nitrogen (BUN), Urinary microalbumin variation.
5) control group (K groups) mouse is fed using normal diet, and injecting normal saline.
6) at the end of modeling the 8th week, 1-2 mouse is selected at random for every group, by these mouse fasting from all groups
Then enough time is put it into the metabolic mouse cage that can acquire urine, collect urine for 24 hours.The urine being collected into is sent to disease
The microalbumin content in room detection urine is managed, CRE and BUN is surveyed with multi-function microplate reader, assesses the degree of kidney injury.
7) H groups and Y group mouse to having generated damage are then begun to and removes intraperitoneal injection CdCl2Outside, also correspond to gavage to
Protection medicine (Astragalus Root P.E) and positive control medicine are given, to equally having generated the M group mouse of damage except intraperitoneal injection CdCl2
Outside, gavage physiological saline is gone back；Gavage is corresponded to the H2 groups and Y2 group mouse that have generated damage and gives protection medicine (astragalus extraction
Object) and positive control medicine, to equally having generated the S group intragastric administration on mice physiological saline of damage；To K group gavage physiological saline；
The gavage time for 4 weeks.
Table 1. is grouped and processing
Three, the measurement of physiological and biochemical index
Ensure constant temperature (indoor temperature is about 25 DEG C), constant humidity after starting modeling, gives 12h dark and the alternate item of illumination
Part supplies ADEQUATE DIET and drinking-water, enable mouse can free feeding water intaking, routine observation dietary amount and amount of drinking water, survey weight change
Change, fasting blood-glucose is measured per weekend, and calculate the average value of data.
After terminating gavage at the 12nd week, the record work of all routines is first carried out, mouse feed is then taken out and it is prohibited
Food measures fasting blood-glucose, and the metabolic mouse cage of urine can be acquired by, which being then placed in, collects urine for 24 hours, is collected into after urine, centrifuges
Removal of impurities, is then stored in -80 DEG C of refrigerators, is used for the detection of urine creatinine and microalbumin.Then the 0.5h abdomens before taking blood
The heparin sodium aqua of chamber injection 1%, eyes-affinity venous blood sampling stand the whole blood being collected into, and low-temperature centrifugation takes supernatant to continue point
Dress, a part is for measuring serum total cholesterol (TC), serum triglyceride (TG), HDL, LDL, blood creatinine, in addition one
Divide for detecting the metabolin ingredient in (LC-MS) blood.In Histomorphological, taking internal organ is dissected, observes internal organs form
Variation, takes kidney to weigh, and calculates kidney weight ratio, then dispenses nephridial tissue, according to paraffin section and ultra-thin cuts before this
The preparation of progress material required by piece and fixed in corresponding fixer；Another part is homogenized for tissue preparation, is made
For detection tissue oxygenization stress enzyme detection Sample storage in -80 DEG C；Remaining part directly freezes in -80 DEG C of refrigerators
In, for immuning tissue's blotting detection and retain spare sample.
Four, result
1, the difference (i.e. M groups and S groups difference) of cadmium induction and STZ induced animal models
Animal model that the diabetic nephropathy model blood glucose rise of STZ inductions is induced compared with cadmium faster, but kidney injury
Index (such as BUN, urine CRE) illustrates that the performance of cadmium induced diabetes nephrosis occurs earlier, and damage becomes apparent from.
Referring to Fig. 1 (A), (B) and (C), by Ultrastructure analysis, the kidney mesentery of cadmium guidance model group (i.e. M groups) is thick
Degree is 244.54nm, and the mesentery thickness of Normal group (i.e. K groups) is 133.70nm, illustrates the kidney of diabetic nephropathy mouse
Phenomena such as appearance glomerular mesangium expansion thickens, and glomerulus hardens, the performance with early diabetic nephropathy, STZ controls
The mesentery thickness of group (i.e. S groups) is only 109nm, illustrates that hyperglycemia can't lead to thickening for kidney mesentery, and Cd and height high in fat
The synergy of sugared feed can then lead to lesion, form diabetic nephropathy.
2, the protective effect of Astragalus Root P.E
Referring to Fig. 1 (D), the kidney mesentery thickness of H groups is 175.35nm, and there is surface Astragalus Root P.E certain protection to imitate
Fruit alleviates the generation of diabetic nephropathy.
Referring to Fig. 2, according to the measurement of mouse fasting blood glucose level, H groups mouse and the comparison of M group mouse, it can be seen that the
Terminate within 12 weeks, Astragalus Root P.E effectively inhibits blood glucose, to maintain the basic activity feature of body.
The toxic effect of cadmium can induced diabetes nephrosis mouse matter and energy metabolic disorder, and Astragalus Root P.E is to blood
The stabilization of fat has certain adjusting protective effect；Referring to Fig. 3, it is logical to body decomposition food acquisition energy can effectively to restore Cd
The protection on road, and be in effective control of high TG present situations to S groups；Referring to Fig. 4, the toxic effect of cadmium being capable of induced diabetes nephrosis
The metabolism of mouse total cholesterol is abnormal, and Astragalus Root P.E has certain adjusting protective effect to the metabolism of total cholesterol,
Illustrate that Astragalus Root P.E has certain protective effect.
Referring to Fig. 5, CRE illustrate be kidney injury degree, as shown, cadmium induction can obviously damage kidney, and
STZ inductions are relatively low for the damage of kidney, and Astragalus Root P.E can effectively alleviate the variation of CRE.
3, it discusses
Although in document report, STZ induced diabetes nephropathy models generally require the longer time, from blood glucose with
And the variation of creatinine level, it is sufficient to show STZ and play inducing action.The diabetic nephropathy mouse model that the present invention is built
Have the characteristics that be different from general diabetic nephropathy model：Cadmium causes diabetic nephropathy to model the time compared to using streptozotocin
(STZ) diabetic nephropathy model built is shorter, and damage becomes apparent from, and is mainly manifested in creatinine (CRE), urea nitrogen (BUN)
And the change of microalbumin occurs earlier, meanwhile, experimental result also indicates that, apparent (Y2 groups and S are acted on positive control drug
Group is compared) unlike, within the model treatment time, pathoglycemia raising caused by Astragalus Root P.E induces STZ does not have
Apparent inhibiting effect (H2 groups are compared with S groups).
4, conclusion
1) long-term Cd exposures can cause body to generate lasting hyperglycemia, eventually lead to the generation of type-2 diabetes mellitus.Sugar
The sick persistent high blood sugar of urine can be with dyslipidemia phenomenon, including free fatty increases, triglycerides increases, and total cholesterol increases
Equal physiology variation.These variations have in generating insulin resistance, the patient for obtaining metabolic syndrome or type-2 diabetes mellitus
Apparent observation.Meanwhile more serious kidney injury (urine CRE reductions, BUN raisings, blood CRE raisings, kidney system are caused
Film thickens).
2) under the induction of Cd long-term damages joint high-sugar-fat-diet, cause insulin sensitivity to reduce, generate insulin and support
It is anti-.Most important two indexs are triglycerides and cholesterol in various lipid materials in blood, are fed with high sugar high in fat, blood
Middle disorders of lipid metabolism.
3) diabetic nephropathy that cadmium combines high sugar structure high in fat is successful, Radix Astragali and its extract to the exposure of long-term cadmium and
The diabetic nephropathy that high-carbonhydrate diet high in fat is caused has the validity of prevention and treatment.
In short, the present invention is small by the diabetic nephropathy for feeding joint intraperitoneal injection caddy induction structure to high sugar high in fat
The changing rule of all kinds of indexs is studied in mouse model, is conducive to the mechanism of action for furtheing elucidate diabetic nephropathy, this hair
Bright to show that DN pathogenesis is complicated, there are multiple links and target spot, single target spot intervention effect is not good enough.And Chinese herbal medicine astragalus is shown
The characteristics of multicomponent and multiple target effect, causes have obvious effect and advantage (for example, with melbine phase in DN preventions in cadmium
Than so that change of blood sugar eases up), be conducive to find the new approach for the treatment of DN, be also beneficial to preferably develop and utilize Radix Astragali.
Claims (8)
1. the application of Radix Astragali or Astragalus Root P.E in preparing the drug for causing diabetic nephropathy for preventing cadmium.
2. application according to claim 1, it is characterised in that：The Astragalus Root P.E is selected from Radix Astragali extractum.
3. application according to claim 2, it is characterised in that：The Radix Astragali extractum is using water to bulk pharmaceutical chemicals Radix Astragali hot dipping
Through concentration and prepared by drying after carrying.
4. application according to claim 1, it is characterised in that：The Radix Astragali or Astragalus Root P.E have stabilizing blood sugar and suppression
The effect of blood glucose rise processed.
5. application according to claim 1, it is characterised in that：The Radix Astragali or Astragalus Root P.E, which have dyslipidemia, to be protected
Shield and adjustment effect.
6. application according to claim 5, it is characterised in that：The dyslipidemia includes total cholesterol and triglycerides liter
It is high.
7. application according to claim 1, it is characterised in that：The Radix Astragali or Astragalus Root P.E, which have kidney injury, to be protected
Shield acts on.
8. application according to claim 7, it is characterised in that：The kidney injury is embodied in urine creatinine reduction, blood
Creatinine increases and kidney mesentery thickens.
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| CN109350691A (en) * | 2018-10-12 | 2019-02-19 | 皖西学院 | A kind of Chinese medicine row's cadmium kidney-tonifying and sperm-producing cream and preparation method thereof |
| CN111920843A (en) * | 2019-05-13 | 2020-11-13 | 北京中医药大学 | Astragalus membranaceus-paecilomyces cicadae fermentation product, and preparation method and application thereof |
| CN114208776A (en) * | 2022-01-27 | 2022-03-22 | 安徽医科大学 | Construction method of male sterility mouse model |
| CN114534808A (en) * | 2022-02-23 | 2022-05-27 | 陕西科技大学 | Microfluidic chip for multi-cell co-culture and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109350691A (en) * | 2018-10-12 | 2019-02-19 | 皖西学院 | A kind of Chinese medicine row's cadmium kidney-tonifying and sperm-producing cream and preparation method thereof |
| CN111920843A (en) * | 2019-05-13 | 2020-11-13 | 北京中医药大学 | Astragalus membranaceus-paecilomyces cicadae fermentation product, and preparation method and application thereof |
| CN114208776A (en) * | 2022-01-27 | 2022-03-22 | 安徽医科大学 | Construction method of male sterility mouse model |
| CN114208776B (en) * | 2022-01-27 | 2022-11-25 | 安徽医科大学 | Construction method of male sterility mouse model |
| CN114534808A (en) * | 2022-02-23 | 2022-05-27 | 陕西科技大学 | Microfluidic chip for multi-cell co-culture and preparation method thereof |
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