CN104352962A - Medicinal composition for treating renal fibrosis, as well as preparation method and application thereof - Google Patents
Medicinal composition for treating renal fibrosis, as well as preparation method and application thereof Download PDFInfo
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- CN104352962A CN104352962A CN201410567744.1A CN201410567744A CN104352962A CN 104352962 A CN104352962 A CN 104352962A CN 201410567744 A CN201410567744 A CN 201410567744A CN 104352962 A CN104352962 A CN 104352962A
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Abstract
The invention provides a medicinal composition for treating the renal fibrosis. The medicinal composition is prepared from the following constituents in parts by weight: 7-13 parts of ginseng, 35-45 parts of radix astragali, 7-13 parts of raw rhubarb, 12-18 parts of herba epimedii, 10-15 parts of angelica sinensis, 7-13 parts of leech, 10-15 parts of curcuma zedoary, 10-15 parts of bombyx batryticatus, and 17-23 parts of oldenlandia diffusa. The invention further provides a preparation method and application of the composition. The medicinal composition disclosed by the invention is rigorous in formula, combined with tonification and purgation, capable of nourishing and loosing bowels with a mutual reason, further can remove turbid and evil, considers about deficiency of qi, slows down development of renal fibrosis, improves the pathological states such as malnutrition and microinflammation for a patient with chronic renal failure, and improves the survival quality of the patient with the chronic renal failure.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition for the treatment of renal fibrosis and its production and use.
Background technology
Renal fibrosis is a kind of pathophysiological change, be kidney function by health to damage, then to damage, until the progressive process of afunction.Kidney is owing to being subject to wound, infection, inflammation, blood circulatory disorder, and the multiple paathogenic factor such as immunoreation stimulates, its intrinsic cell damage, developing into the later stage there is a large amount of collagen deposition and gathering, cause excess of the kidney matter to harden gradually, form cicatrix, until kidney completely loses organ function.Intrinsic cell fibrosis, the process of sclerosis i.e. the process of renal fibrosis in kidney.Renal fibrosis is feature with the abnormal deposition of extracellular matrix (ECM).
Traditional Chinese Medicine, without renal fibrosis one word, according to its clinical manifestation, generally belongs to " edema ", " asthenia ", " difficulty in urination ", " obstruction and rejection " category, pathomechanism is that function is impaired, the positive deficiency of vital energy declines, and wet, expectorant, the stasis of blood, poison are kept in body, define in kidney " mass in the abdomen ".Long-pending is body inner product block, have the advantages that tangible and hard work does not move, show extracellular matrix build-up during chronic kidney disease progress, glomerular capsule is adhered, blood vessel loop is inaccessible, and FSGS, kidney region fibrosis all can belong to kidney Nei “ mass in the abdomen " category.
Summary of the invention
The object of the present invention is to provide a kind of pharmaceutical composition for the treatment of renal fibrosis.Another object of the present invention is to preparation method and purposes that said composition is provided.
Particularly, the invention provides a kind of pharmaceutical composition for the treatment of renal fibrosis, its raw material is containing, for example the component of lower weight proportion:
Further, its raw material is containing, for example the component of lower weight proportion:
Further, its raw material is made up of the component of following weight proportion:
Further, its raw material is made up of the component of following weight proportion:
For above-mentioned composition flavour of a drug proportioning, the present invention it may be noted that:
(1) in preclinical practice, compared for Radix Panacis Quinquefolii and Radix Ginseng to the impact of compositions, when finding to use Radix Ginseng, compositions curative effect is better than Radix Panacis Quinquefolii.Its reason may be, the traditional Chinese medical science thinks Radix Ginseng partial heat, and focus on QI invigorating, Radix Panacis Quinquefolii is put down cooler, can nourishing YIN to lower pathogenic fire, and Chronic Renal Failure Patients deficiency in origin mostly is the deficiency of vital energy, therefore uses ginseng qi-tonifying kidney tonifying.
(2) in early-stage Study, use Radix Salviae Miltiorrhizae is circulation promoting vein relaxing medicine, but after replacing Radix Salviae Miltiorrhizae with Radix Angelicae Sinensis Bombyx Batryticatus, compositions curative effect is better.Its reason may be, with the passing of time Chronic Renal Failure Patients is converted to yang deficiency by the deficiency of vital energy, and Radix Salviae Miltiorrhizae is cold, and Radix Angelicae Sinensis is then warm in nature, and promoting blood circulation to remove obstruction in the collateral curative effect is more excellent; Bombyx Batryticatus salty in the mouth is pungent, kind dissipating phlegm and resolving masses, and chronic kidney hypofunction with the passing of time, keep in body, forms obstinate “ mass in the abdomen in kidney by wet, expectorant, the stasis of blood, poison ", add Bombyx Batryticatus and then strengthen Xiao Disorder effect of invigorating blood circulation.
(3) consumption proportion of each taste medicine in pharmaceutical composition of the present invention, be that inventor's treatment of summing up out in clinical practice is for a long time early stage, mid-term Chronic Renal Failure Patients curative effect the best proportioning, and when roll up separately wherein QI invigorating medicine or activating blood and removing stasis drug or heat-clearing and toxic substances removing medicine ratio time, patient's curative effect weakens.
According to long-term clinical practice, in the final resulting composition of the present invention, Radix Ginseng, the Radix Astragali are monarch, and kidney is the residence of extreme misery, contains nephroyin and nephroyang, vigour virtual loss in kidney, directly need the large agent Radix Astragali, Radix Ginseng strongly invigorating primordial QI basic to reinforce the kidney; Herba Epimedii assists monarch drug warming and recuperating the kidney-YANG to be minister; Radix Angelicae Sinensis, Rhizoma Curcumae, Hirudo, Bombyx Batryticatus are invigorated blood circulation Xiao Disorder, and Herba Hedyotidis Diffusae, Radix Et Rhizoma Rhei removing toxic substances let out and turbidly to help for assistant, gather altogether reinforce the kidney Xiao Disorder, detoxify and let out turbid merit.Herba Epimedii and Herba Hedyotidis Diffusae compatibility, for the pathogenesis of this disease " deficiency in origin and excess in superficiality, coldheat complex ", embody the Important Thought of " simultaneously using both cold and hot drugs ".Full side " simultaneously using both cold and hot drugs, reinforcement and elimination in combination ", compatibility is precise and appropriate.
Wherein, it be by the medicated powder of the raw material of described proportioning, water extract or/and ethanol extraction is active component, add the dosage form that pharmaceutically conventional adjuvant or complementary composition are prepared from.
Water extract or be used as medicine with medicated powder is all Chinese medicine tradition occupation modes, and after water extraction, because the soluble end of water is wide, can, by most of effective ingredient stripping, medicine is more easily absorbed by the body, onset be faster, the form of medication such as such as decoction; Be used as medicine with former powder, the surface area of medicated powder is larger, also effective ingredient absorption in vivo in medical material is conducive to, but medical material un-extracted, effective ingredient still needs stripping in vivo to absorb again, and the relative water extract of its onset is comparatively slow, but also weakens the toxicity that in medical material, harmful components cause human body simultaneously, be suitable for long-term taking, as former powder is prepared into the form of medication such as pill.At present in pharmacy procedure, ethanol extracts medicine as solvent, also be one of the most common extracting mode, ethanol is semi-polarity solvent, solubility property circle is between polarity and non-polar solven, some composition water miscible can be dissolved, also some compositions that non-polar solven dissolves can be dissolved, usually decocting is replaced with ethanol extraction, thus avoid the stripping of a large amount of invalid components, improve concentration and the extraction efficiency of effective ingredient, but comparatively water is expensive for the price of ethanol, in the large production of modern pharmaceutical industry, in order to save production cost, usually still based on decocting.
Pharmaceutically acceptable adjuvant of the present invention, refer to the material be included in addition to the active ingredient (s in dosage form, include but are not limited to filler (diluent), lubricant (fluidizer or antitack agent), dispersant, wetting agent, binding agent, regulator, solubilizing agent, antioxidant, antibacterial, emulsifying agent, disintegrating agent etc.Binding agent comprises syrup, arabic gum, gelatin, sorbitol, tragacanth, cellulose and its derivates (as microcrystalline Cellulose, sodium carboxymethyl cellulose, ethyl cellulose or hydroxypropyl methylcellulose etc.), gelatine size, syrup, starch slurry or polyvinylpyrrolidone etc.; Filler comprises lactose, Icing Sugar, dextrin, starch and derivant thereof, cellulose and its derivates, inorganic calcium salt (as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate etc.), sorbitol or glycine etc.; Lubricant comprises micropowder silica gel, magnesium stearate, Pulvis Talci, aluminium hydroxide, boric acid, hydrogenated vegetable oil, Polyethylene Glycol etc.; Disintegrating agent comprises starch and derivant (as carboxymethyl starch sodium, Explotab, pregelatinized Starch, modified starch, hydroxypropyl starch, corn starch etc.), polyvinylpyrrolidone or microcrystalline Cellulose etc.; Wetting agent comprises sodium lauryl sulphate, water or alcohol etc.; Antioxidant packages is containing sodium sulfite, sodium sulfite, sodium pyrosulfite, dibutyl benzoic acid etc.; Antibacterial comprises 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol etc.; Regulator comprises hydrochloric acid, citric acid, potassium hydroxide (sodium), sodium citrate and buffer agent (comprising sodium dihydrogen phosphate and sodium hydrogen phosphate) etc.; Emulsifier package is smooth containing Tween-80, fatty acid Pyrusussuriensis, pluronic gram F-68, lecithin, fabaceous lecithin etc.; Solubilizing agent comprises tween 80, bile, glycerol etc.
Described pharmaceutically acceptable complementary composition, it has certain physiologically active, but adding of this composition can not change above-claimed cpd or the leading position of derivant in treatment of diseases, and only play auxiliary effect, these auxiliary effects are only the utilizations to this composition known activity, are the usual adjuvant treatment modality of field of medicaments.If by above-mentioned complementary composition and the compounds of this invention with the use of, still should belong to the scope of protection of the invention.
Further, described dosage form is through gastrointestinal absorption dosage form.
Present invention also offers the preparation method of aforementioned pharmaceutical compositions, it comprises following operating procedure:
(1) raw material is taken by proportioning;
(2) get Herba Epimedii, Hirudo, pulverize, use 60%v/v ethanol extraction, extracting solution reclaims ethanol, is condensed into fluid extract; Medicinal residues and the Radix Astragali, Radix Angelicae Sinensis, Rhizoma Curcumae, Bombyx Batryticatus, Herba Hedyotidis Diffusae 5 taste medicine extracting in water, filter, filtrate reduced in volume, leaves standstill, get supernatant and add above-mentioned fluid extract, after concentrating under reduced pressure, add Radix Ginseng, Rhubarb, be prepared into preparation.
Alcohol extraction procedure of the present invention, can adopt ultrasonic, backflow, dipping, percolation etc.
Water extraction method of the present invention, can adopt ultrasonic, decoction, warm macerating etc.
Present invention also offers the purposes of aforementioned pharmaceutical compositions in the medicine of preparation treatment renal fibrosis
Present invention also offers aforementioned pharmaceutical compositions and prepare the purposes in the medicine improving Maintenance Hemodialysis Patients Micro-inflammation state, treatment renal interstitial fibrosis.
Pharmaceutical composition of the present invention, prescription is rigorous, full side's reinforcement and elimination in combination, there is benefit to have to rush down, reinforcing and reducing phase factor, not only remove turbid pathogen but also looked after deficiency of vital energy assertive evidence, both can delay renal fibrosis progress, also can improve the pathological states such as Chronic Renal Failure Patients malnutrition, micro milling, improve the life quality of Chronic Renal Failure Patients.
Below in conjunction with detailed description of the invention, the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on content of the present invention all belong to scope of the present invention.
Detailed description of the invention
Embodiment 1
Above-mentioned raw materials, decocts with water, and merges decocting liquid.Every day potion, decocting 400ml, oral, sooner or later twice sub-service.
Embodiment 2
Above-mentioned raw materials, decocts with water, and merges decocting liquid, after concentrated, add appropriate amount of starch and dextrin, prepares granule.
Embodiment 3
Herba Epimedii, Hirudo powder are broken into coarse powder, with 8 times amount 60% alcohol dipping 12 hours, percolation, collect percolate, reclaim ethanol, be condensed into fluid extract.Medicinal residues and the Radix Astragali, Radix Angelicae Sinensis, Rhizoma Curcumae, Bombyx Batryticatus, Herba Hedyotidis Diffusae 5 taste medicine add recipe quantity 8 times of water soakings 1 hour, decoct 3 times, each 1 hour, collecting decoction, and filter, filtrate reduced in volume is to relative density (1.02-1.05g/cm
3, 80 DEG C), leave standstill 12 hours, get supernatant and add above-mentioned fluid extract, be evaporated to relative density (1.20-1.25g/cm
3, 80 DEG C) extractum.Add Radix Ginseng, Radix Et Rhizoma Rhei fine powder, granulate with dextrin, granulate, subpackage both obtained, and 10g granule is equivalent to 21.593g raw medicinal herbs.
Embodiment 4
Above-mentioned raw materials, with 80 ~ 85%w/w alcohol reflux 2 times, alcohol extract is for subsequent use, and medicinal residues decoct with water 2 times, merges decocting liquid.Alcohol extract merges with decocting liquid after reclaiming ethanol, after concentrated, add appropriate microcrystalline Cellulose, spraying dry, prepares capsule.
Beneficial effect of the present invention is illustrated below by way of test example.
Test example 1 pharmaceutical composition of the present invention is on the impact of Maintenance Hemodialysis Patients Micro-inflammation state
1 case-data
All cases all derive from certain hospital outpatient Maintenance Hemodialysis Patients, and totally 40 examples, are divided into two groups by table of random number.
Treatment group 20 example, wherein man 11 example, female 9 example, 51.34 ± 1.3 years old mean age, dialysis ages is the shortest 6 months, the longest 72 months, average 31.42 ± 4.2 months, and protopathy is chronic glomerulonephritis 12 example, diabetic nephropathy 5 example, hypertensive nephropathy 1 example, urate nephropathy 1 example, polycystic kidney 1 example.
Matched group 20 example, wherein man 10 example, female 10 example, 52.75 ± 1.2 years old mean age, dialysis ages is the shortest 6 months, the longest 74 months, average 32.18 ± 3.89 months, and protopathy is chronic nephritis 11 example, diabetic nephropathy 5 example, hypertensive nephropathy 1 example, urate nephropathy 1 example, polycystic kidney disease: 2 cases.Two groups of data sexes, age, dialysis ages, protopathy no significant difference, have comparability.
Healthy group 20, is the Healthy People of my Out-patient Department health check-up.Wherein man 10, female 10,50.32 ± 1.68 years old mean age.
2 Therapeutic Method
Treatment group and matched group all give Primary Care, i.e. high-quality protein diet, control blood pressure, blood glucose, infection, anemia, correct calcium and phosphorus metabolism disorder.Two groups all adopt treasure AK200s dialysis machine, bicarbonate dialysis liquid system, polysulfone membrane dialyser, area 1.4m
2, Kt/V maintains > 1.2, each dialysis 4 hours, and dialysate flow 500ml/min, blood flow 200-250ml/min, dialyse 3 times weekly, the March course for the treatment of.Treatment group on the basis of the above, adds medicament composition granule agent of the present invention 10 grams (10g granule is equivalent to 21.593g raw medicinal herbs), every day three times, continuous March.
3 observation items and Testing index
Two groups of patients are before treatment and treat and observe for latter 3 months.
3.1 nutritional status monitorings
3.1.1 anthroposomatology index
Dry weight, triceps skinfold thickness (TSF), upper-arm circumference (AC), arm muscle circumference (AMC) after height, dialysis.AMC(cm)＝AC-3.14×TSF(cm)。Dialysis to terminate in rear 10-20min arm measure TSF, AC on MHD patient's internal fistula offside.
3.1.2 blood biochemistry index
Detect albumin (Alb), prealbumin (PA), T-CHOL (TC), triglyceride (TG), HDL-C (HDL-C), low-density lipoprotein cholesterol (LDL-C), hemoglobin (Hb).
3.2 inflammation index
Phlebotomize and look into C reactive protein (CRP), tumor necrosis factor (INF-α), interleukin-6 (IL-6).Normal healthy controls group is adopted limosis vein blood and is looked into These parameters.CRP adopts immunoturbidimetry, INF-α, IL-6 ELISA method.
4 statistical methods
Enumeration data represents with (X ± s), adopts the capable t inspection of SPSS11.0 software or variance analysis.
5 results
5.1 anthroposomatology indexs
After treating March, treatment group dry weight, MAMC obviously raise (P<0.05) before comparatively treating; Though TSF has rising, not statistically significant (P ﹥ 0.05).Matched group indices is unchanged.Treatment group compares with matched group, and difference has statistical significance (P<0.05).In table 1.
The change (X ± s) of anthroposomatology index before and after table 1 medication
Note: compare with normal healthy controls group,
*p<0.05; Compare with before treatment
△p<0.05; Compare with matched group
#p<0.05.
5.2 blood biochemistry index change treatment group Alb, PA, Hb raise obviously after treating March, statistical significance (P<0.05) is had with comparing difference before treatment, control group A lb, after the PA3 month without significant change, rise to some extent before Hb comparatively treats, but no significant difference (P ﹥ 0.05).Obvious afterwards with matched group comparison therapy group Alb, PA, Hb3 month
onrise, difference has statistical significance (P<0.05).No significant difference (P<0.05) before and after two groups of blood TC, TG, HDL-C, LDL-C horizontal stretchers.In table 2.
Nutritive index change (X ± s) before and after table 2 medication
Table 2 continues
Note: compare with normal healthy controls group,
*p<0.05; Compare with before treatment
△p<0.05; Compare with matched group
* #p<0.05
After 5.3MHD patient's inflammation index change treatment group treats March, CRP, INF-α, IL-6 are all lower than before treatment (P<0.05), matched group CRP, INF-α, IL-6 are all higher than before treatment, but no significant difference (P>0.05).Compare between treatment group with group after treatment of control group, treatment group CRP, INF-α, IL-6 are all lower than matched group (P<0.05).In table 3.
Front and back inflammation index change (X ± s) treated by table 3
Note: compare with normal healthy controls group,
*p<0.05; Compare with before treatment
△p<0.05; Compare with matched group
#p<0.05
6 discuss
Micro-inflammation state refers to that patient does not have whole body or locally dominant clinical infection sign, but there is low-level, lasting inflammatory conditions, show as inflammatory factor, as tumor necrosis factor (INF-α), interleukin-6 (IL-6), interleukin-1 (IL-1) and inflammation related proteins, as High-sensitivity C reactive protein (hs-CRP), Fibrinogen, serum amyloid protein slightly raise.Stenvinkl etc. propose Chronic Renal Failure Patients and there is malnutrition-inflammation-atherosclerosis (MIA) syndrome, and inflammation is key link wherein, are cause malnutrition and atherosclerotic key factor.CRP level, malnutrition, atherosclerosis are the strong predictor affecting dialysis patient prognosis, there is obvious dependency in CRP level and malnutrition, atherosclerosis, therefore Micro-inflammation state becomes the key that MHD patient prevents cardiovascular complication.CRP is a kind of acute phase protein, and be the most responsive and special index of chronic inflammatory disease, hs-CRP can detect low-level CRP concentration, is the Sensitive mark of low-level inflammation; IL-6 is important inflammatory mediator, stimulates liver to produce CRP.TNF-α produces primarily of the mononuclear phagocyte of activation, is that one has multiple bioactive polypeptide cytokines, closely related with the immunoreation of body, inflammatory reaction, brings out systemic inflammatory response syndrome by active cell cytokine network system.This is studied in 40 routine patients has 31 examples serum CA125 before treatment namely to raise, account for 77.50% of patient's number, consistent with bibliographical information, and normal healthy controls group CRP only has 10% (2 example) to exceed normal value, illustrates that Maintenance Hemodialysis Patients exists Micro-inflammation state really.After this research matched group patients with hemodialysis, CRP, TNF-a, IL-6 raise before all comparatively treating, and illustrate that in hemodialysis, monokaryon one macrophage is activated, a large amount of cytokine activation, IL-6, TNF-a regulate acute-phase response as the proinflammatory factor.
There is the malnutrition of two types in uremic patient, I type malnutrition is reduced by insufficient, the appetite of dialysing and causes albumen and Energy intaking deficiency to cause, and II type malnutrition is relevant to inflammation.Inflammatory factor increases the metabolism of muscle protein, makes body weight loss, occurs becoming thin, and serum albumin synthesis reduces and causes hypoalbuminemia.Uremic patient usually 2 kinds of malnutrition exists simultaneously.Therefore, improve dialysis adequacy in chronic and extra-nutrition material, improve the strategy that Micro-inflammation state is treatment.After treatment group takes pharmaceutical composition of the present invention, average dry weight rises to (57.82 ± 10.25) kg, MAMC meansigma methods by (55.32 ± 12.68) kg before treating and rises to (21.56 ± 4.83) cm by (17.78 ± 6.12) cm before treating.Obviously raise after Alb, PA, Hb treatment in blood biochemistry index, These parameters is improved may improve patient's appetite with pharmaceutical composition of the present invention, and albumen is taken in be increased, and aminoacid reduces relevant with Protein catabolism.After treatment group gives medicine composite for curing of the present invention, CRP, TNF-a, IL-6 reduce before all comparatively treating, showing that nutriture improves can enhancing human body immunity function, developing of defence micro milling, and alleviating of micro milling makes patient's appetite strengthen, the nutrient substance such as protein, carbohydrate is taken in be increased, nutriture is impelled to take a turn for the better, the two is mutually promoted, interacts, the progress of common control MHD patient malnutrition and micro milling, reduce cardiovascular event to occur, improve life in patients, extend life cycle.
The experimentation of the anti-renal interstitial fibrosis rat of test example 2 pharmaceutical composition of the present invention
1 experiment material
1.1 Experimental agents
Pharmaceutical composition of the present invention, prepares by embodiment 1, and 1ml is containing crude drug 1.88 grams.Benazepril every sheet content 10mg, Novartis Pharma AG produces.
1.2 experiment reagent
Rabbit Chinese People's Anti-Japanese Military and Political College Mus TGF-β
1polyclonal antibody test kit, rabbit Chinese People's Anti-Japanese Military and Political College Mus Fibronectin polyclonal antibody test kit, goat anti-rabbit igg are produced by Wuhan doctor's moral company, and rabbit Chinese People's Anti-Japanese Military and Political College Mus type III collagen polyclonal antibody test kit is purchased from Beijing Zhong Shan Bioisystech Co., Ltd.
1.3 major experimental instruments
CD-6 multi-media image analytical system, OLYMPUS BX-50400 times optical microscope.
1.4 laboratory animal
Healthy SD female rats 36, body weight 180 ± 20 grams, Luzhou Medical College Experimental Animal Center provides.
2 experimental techniques
2.1 modelling
Aseptically, by 36 rat skin lower injection ketalar (150mg/kg) anesthesia, skin is cut in left side midriff, free left side ureter, except sham operated rats not ligation ureter is sewed up except abdominal cavity, all the other respectively group respectively in renal pelvis place and ureter 1/3 place with after silk thread ligation, cut off ureter, layer-by-layer suture skin, namely prepares UUO model.
2.2 animal process
After laboratory animal adaptability feeds one week, be divided into 4 groups of (1) sham operated rats, 6 (2) model group 10 (3) pharmaceutical composition groups of the present invention, 10 (4) benazepril group 10 at random by random alignment table.Sham operated rats and model group clean tap water gavage every day 1 time, medicine group, in preoperative 1 day, gives pharmaceutical composition 12.45ml.kg of the present invention postoperative every day respectively
-1.d
-1, benazepril suspension 10mg.kg
-1.d
-1gavage, postoperative 2 weekends put to death each group of rat, often organize to select 6 rats to get left kidney at random to do indices inspection.
2.3 methods of sample collection
After sacrifice of animal, take out left kidney immediately, fix 24 hours by 10% neutral formalin, through dewatering, embedding, be cut into 4 micron thickness, make HE dyeing and immunohistochemical staining.
2.4 renal pathology inspections
2.4.1 immunohistochemical staining
SABC method: 1 anti-(the TGF-β that (removing endogenous catalase active) (3) 1% normal sheep serums closed (4) dropping that the conventional aquation (2) that dewaxes of the paraffin section of (1) 4 micron thickness immerses 3% hydrogen peroxide 5 minutes is diluted
1type III collagen, Fibronectin polyclonal antibody, dilution factor is 1:100), 4 DEG C are spent the night, and PBS rinses 3 times (5) and drips excessive goat anti-rabbit igg, room temperature 1 hour, PBS rinses 3 times (6) and drips SABC complex, room temperature 1 hour, and PBS rinses that (7) DAB develops the color, haematoxylin is redyed, dewaters, transparent, sealing.Replace primary antibodie as negative control using PBS.
2.4.2 renal tubular interstitium pathological change
Renal tubulointerstitial lesion degree is classified with reference to domestic Tang Zheng, is divided into III level, i.e. I level: tubulo-interstital pathological changes is dispersed in slightly, range L EssT.LTssT.LT15%; II level: pathological changes is stove or small pieces distribution, scope >=15%, <50%; III level: pathological changes in the form of sheets or fill the air distribution, scope >=50%.
2.4.3 pathology image analysis
After immunohistochemical staining, microscopy excess of the kidney matter and the interior brown granular of Interstitial cell slurry are positive.Often open section and get 5 width figure at renal interstitial, coloration result Quantitative Pathologic Image Analysis system carries out semi-quantitative analysis, and renal interstitial positive expression area and renal interstitial area ratio are renal interstitial positive expression rate, 5 result computation of mean values and standard deviation.
2.4.5 statistical procedures
Experimental data represents with × ± S, and measurement data group difference adopts one factor analysis of variance q inspection, and numeration data adopts the rank test by grade classified data, with the process of SPSS10.0 statistical software.
3 results
3.1 pathological observation
Perusal sham operated rats nephridial tissue is ruddy without enlargement, and model group, pharmaceutical composition group of the present invention, benazepril group sick side Kidney Volume obviously increases, and color bleaches, and cortex is thinning.HE coloration result, rats in sham-operated group nephridial tissue structure is without exception.The atrophy of model group renal cells, degeneration, necrosis, come off, the cystic dilatation of part tube chamber, renal interstitial monokaryon, lymphocytic infiltration and fibrous tissue are formed.Pharmaceutical composition group of the present invention changes all to have compared with model group with benazepril group sick side Pathological and alleviates in various degree.
3.2 each treated animal renal interstitial fibrosis degree: (see table 4)
Table 4 is group tubulointerstitial fibrosis in rats fibrosis (x ± s) respectively
Note: compare with model group,
*p<0.05; Compare with pharmaceutical composition group of the present invention
#p>0.05.
As shown in Table 4, sham operated rats is without renal tubulointerstitial lesion.All there is renal interstitial fibrosis in model group, pharmaceutical composition group of the present invention, benazepril group, pharmaceutical composition group of the present invention, benazepril group renal interstitial fibrosis degree comparatively model group obvious difference (
*p<0.05); Two treatment group fibrosis there was no significant differences (
#p>0.05).
3.3III Collagen Type VI, Fibronectin, TGF-β
1expression: (see table 5)
Table 5 is group kidney of rats interstitial ColIII, FN, TGF-β respectively
1area ratio (%) change (x ± s)
Note: compare with sham operated rats,
*p<0.05; Compare with model group,
#p<0.05; Compare with pharmaceutical composition group of the present invention,
▲p>0.05 (lower same)
Model group, pharmaceutical composition group of the present invention, benazepril group renal interstitial ColIII, FN, TGF-β as shown in Table 5
1area ratio increase, have compared with matched group significant difference (
*p<0.05); Compared with model group, two medicine group renal interstitial ColIII, FN, TGF-β
1expression weakens, area ratio reduces (
#p<0.05); Pharmaceutical composition group of the present invention and benazepril group area ratio no significant difference (
▲p>0.05).
4 discuss
Renal tubulointerstitial fibrosis is the progressive common pathological characters proceeding to end-stage renal failure of various chronic renal disease.The mechanism that interstitial fibrosis causes glomerular filtration rate to decline is: between tubule, blood capillary is narrow, and vascular resistance increases, and causes glomerule blood flow and declines; Renal tubules, particularly proximal tubular atrophy consequential damage renal tubular function, affects glomerular filtration rate by bulb feedback mechanism; Interstitial disease of renal tubule also directly can cause glomerular sclerosis or fibrosis crescent pathological changes.Renal interstitial fibrosis shows as the deposition of extracellular matrix (ECM) compositions such as interstitial collagen (I type, II type, type III), fibronectin (FN) and some glycoproteins at kidney, cause normal Reno-colic fistula to change, renal function is faded and is lost.ECM is in continuous metabolic turnover and degrades in the dynamic equilibrium reinvented under normal circumstances, and when this dysequilibrium, ECM will in renal interstitial over-deposit.Type III collagen, Fibronectin are the main components of ECM, the faint expression of the rarely seen renal interstitial of normal rat, during kidney injury, the activation of renal interstitial fibroblast, propagation, type III collagen, Fibronectin are expressed at renal interstitial obviously to be increased, therefore, detect type III collagen, Fibronectin at the expression of renal interstitial, kidney region fibrosis degree can be reflected.TGF-β thinks most important fibrosis somatomedin at present.It stimulates ECM to produce and deposition by following four aspects: (1) induction I, III, IV, collagen type v, FN, LN transcribe, synthesize and secrete.(2) suppress the expression of metalloproteases (MMP), promote PAI and TIMP synthesis, thus reduce ECM degraded.(3) cell surface ECM receptor is promoted---integrin number increases, and makes sticking between cell and substrate strengthen and impel apposition.(4) it or fibroblastic chemotactic factor, and stimulate fibroblast proliferation.TGF-β synthesizes more at the stronger then ECM of the expression of renal interstitial, imply that kidney region fibrosis is more serious.Therefore, block or reduce the expression of TGF-β, contribute to the renal interstitial fibrosis that slows down.This experiment shows that pharmaceutical composition of the present invention can reduce renal interstitial TGF-β
1, ColIII, FN expression, inferring that it can cause the generation of the fine factor by blocking, reducing the synthesis of extracellular matrix, thus reaching the effect of anti-kidney region fibrosis.
Claims (10)
1. treat a pharmaceutical composition for renal fibrosis, it is characterized in that: its raw material is containing, for example the component of lower weight proportion:
Radix Ginseng 7-13 part Radix Astragali 35-45 part Radix Et Rhizoma Rhei 7-13 part Herba Epimedii 12-18 part Radix Angelicae Sinensis 10-15 part Hirudo 7-13 part Rhizoma Curcumae 10-15 part Bombyx Batryticatus 10-15 part Herba Hedyotidis Diffusae 17-23 part.
2. pharmaceutical composition according to claim 1, is characterized in that: its raw material is containing, for example the component of lower weight proportion:
Radix Ginseng 10 parts of Radixs Astragali 40 parts of Radix Et Rhizoma Rhei 10 parts of Herba Epimedii 15 parts of Radix Angelicae Sinensis 12 portions of Hirudos 10 parts of Rhizoma Curcumae 12 parts of Bombyx Batryticatus 12 parts of Herba Hedyotidis Diffusaes 20 parts.
3. pharmaceutical composition according to claim 1, is characterized in that: its raw material is made up of the component of following weight proportion:
Radix Ginseng 7-13 part Radix Astragali 35-45 part Radix Et Rhizoma Rhei 7-13 part Herba Epimedii 12-18 part Radix Angelicae Sinensis 10-15 part Hirudo 7-13 part Rhizoma Curcumae 10-15 part Bombyx Batryticatus 10-15 part Herba Hedyotidis Diffusae 17-23 part.
4. pharmaceutical composition according to claim 3, is characterized in that: its raw material is made up of the component of following weight proportion:
Radix Ginseng 10 parts of Radixs Astragali 40 parts of Radix Et Rhizoma Rhei 10 parts of Herba Epimedii 15 parts of Radix Angelicae Sinensis 12 portions of Hirudos 10 parts of Rhizoma Curcumae 12 parts of Bombyx Batryticatus 12 parts of Herba Hedyotidis Diffusaes 20 parts.
5. the pharmaceutical composition according to Claims 1 to 4 any one, it is characterized in that: it be by the medicated powder of the raw material of described proportioning, water extract or/and ethanol extraction is active component, add the dosage form that pharmaceutically conventional adjuvant or complementary composition are prepared from.
6. pharmaceutical composition according to claim 5, is characterized in that: described dosage form is through gastrointestinal absorption dosage form.
7. the preparation method of pharmaceutical composition described in Claims 1 to 4 any one, is characterized in that: it comprises following operating procedure:
(1) raw material is taken by proportioning;
(2) get each raw material, extracting in water, merge Aqueous extracts, preparation.
8. preparation method according to claim 7, is characterized in that: in water extraction process, under after Radix Et Rhizoma Rhei.
9. the purposes of pharmaceutical composition described in claim 1 ~ 6 any one in the medicine of preparation treatment renal fibrosis.
10. pharmaceutical composition described in claim 1 ~ 6 any one improves the purposes in the medicine of Maintenance Hemodialysis Patients Micro-inflammation state, treatment renal interstitial fibrosis in preparation.
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| CN107582648A (en) * | 2017-10-18 | 2018-01-16 | 华中科技大学同济医学院附属协和医院 | It is a kind of to be used to treat Chinese medicine preparation of CKD and preparation method thereof |
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| 张琼等: "扶肾泄浊法对维持性血液透析患者微炎症状态的影响", 《泸州医学院学报》 * |
| 王虎: "肾衰丸中大黄素的含量测定", 《中国医院药学杂志》 * |
| 蒲清荣等: "肾纤康颗粒超声提取工艺研究", 《现代医药卫生》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106697A (en) * | 2015-09-07 | 2015-12-02 | 江志鑫 | Pharmaceutical composition for resisting renal fibrosis and application of pharmaceutical composition |
| CN107582648A (en) * | 2017-10-18 | 2018-01-16 | 华中科技大学同济医学院附属协和医院 | It is a kind of to be used to treat Chinese medicine preparation of CKD and preparation method thereof |
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