CN107028938A - Application of the lipoamide in delaying and treating diabetic nephropathy - Google Patents
Application of the lipoamide in delaying and treating diabetic nephropathy Download PDFInfo
- Publication number
- CN107028938A CN107028938A CN201710249412.2A CN201710249412A CN107028938A CN 107028938 A CN107028938 A CN 107028938A CN 201710249412 A CN201710249412 A CN 201710249412A CN 107028938 A CN107028938 A CN 107028938A
- Authority
- CN
- China
- Prior art keywords
- lipoamide
- purposes
- kidney
- composition
- preparing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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
- A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
- A61K31/385—Heterocyclic compounds having sulfur as a ring hetero atom having two or more sulfur atoms in the same ring
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a kind of lipoamide for preparing the purposes in delaying and treating the composition of diabetic nephropathy.
Description
Technical field
The invention belongs to pharmaceutical technology field, lipoamide is related in particular in delaying and treating diabetic nephropathy
Purposes.
Background technology
The incidence of disease of diabetes is constantly rising, the incidence of disease of one of severe complication of diabetes --- diabetic nephropathy
Also it is in raise trend year by year, it has also become the one of the main reasons for causing the Etiological and diabetes of chronic renal failure lethal.Greatly
It there are about 30% type 1 diabetes patient and 25%~40% diabetes B patient understand complicated with diabetes nephrosis.However, glycosuria
The pathogenesis of sick nephrosis is sufficiently complex, and current result of study points out metabolic disorder, hemodynamic responses, inflammatory reaction machine
The many factors such as system, cell factor, oxidative stress, inherent cause, kinin system and autophagy take part in the morbidity of diabetic nephropathy,
But its precise mechanism is not yet clear and definite, therefore it is always more intractable difficult medical problem to the treatment of diabetic nephropathy, lacks always
Effective medicine.Conventional treatment thoughts are concentrated mainly on the peripheral hazards such as control blood glucose, blood pressure, blood fat, but control
Therapeutic effect is unsatisfactory.Therefore, look for prevention and prevent the effective ways of renal function progressive decline, delay and treat diabetes
Nephrosis, very urgent task is prevented and treated as current diabetic nephropathy.
Pathogenesis of Diabetic Nephropathy and pathological manifestations are more complicated, the main fibre including glomerulosclerosis and renal tubular interstitium
Dimensionization.Renal interstitial fibrosis shows as the extracellular matrix that normal renal tubule and renal interstitial structure are largely assembled
(ECM) replace, myofibroblast occur in interstitial.Renal cells (RTECs) is substantially thin as main kidney simultaneously
, there is renal cells to myofibroblast transdifferentiation (EMT) in born of the same parents.Myofibroblast has very strong secretion ECM
Ability, while also release suppresses the enzyme that ECM is decomposed, decompose ECM and reduce and accumulate, directly result in the fibrosis of interstitial with
The damage of renal function.
Transforminggrowthfactor-β1 (TGF-β 1) be acknowledged as diabetic nephropathy tissue fibrosis development it is key it is pathogenic because
Son, it is played by downstream serine/threonine kinases receptors (Smad) signal path in diabetic nephropathy progressive pathological development
Important function.New albumen N (SnoN) related Ski is one of Ski proto-oncogene family members, and its original research concentrates on embryo
In terms of fetal hair is educated with oncogenicity effect.Recent studies have found that SnoN is consideration convey record suppression important in TGF-β 1/Smad signal paths
The factor processed.Its by with TGF-β1The Smads protein-interactings in signal downstream, negative regulation TGF-β1Signal path, plays blocking
The effect of progression of fibrosis.Inventor once in diabetic nephropathy pathogenic process SnoN express change studied (《In
State's pathologic, physiologic magazine》, the phase of volume 24 6, page 1188 in 2008), find as diabetic nephropathy course advancement SnoN expression subtracts
It is few, and high sugar stimulation can cause rat primary renal cells SnoN protein expressions to reduce, and in time dependence, show
The expression reduction of SnoN protein levels is the important pathogenesis of diabetic nephropathy kidney fibrosis.Therefore increased by means such as medicines
Plus SnoN expression is extremely important for intervening diabetic nephropathy kidney fibrosis process.But the medicinal application of current this respect is very
It is limited.
On the other hand, if can act on by medicine increases the stability of SnoN albumen, it perhaps will be helpful to intervene diabetes
Nephrosis progression of fibrosis.But do not reported that medicine was for the intervention of SnoN protein stabilities and further fine to kidney so far
The influence of dimensionization.
Regulation and control (the J Biol Chem.2007 volumes 13,9475 of kinases (TAK-1) are also activated by transforming growth factor β
Page).TAK1 be MAPK kinases (Mitogen-Activated Protein Kinase Kinase,
MAPKK) one of family member, can be by the phosphorylation activation of TGF-β 1.The research of the past finds the progress damaged with Pathological,
The expression of TAK1mRNA and albumen and activity increase, and renal tubular epithelial can be mediated to occur Phenotypic Change.And apply TAK1 suppressions
Preparation for treating db/db mouse can substantially reduce Urine proteins, mitigate pathological change, reduce macrophages infiltration etc..But in the past
Do not have to report in document SnoN protein stabilities and renal cells to myofibroblast transdifferentiation (EMT) it
Between relation, do not have yet
Alpha-lipoic acid is that containing the pentacyclic fat-soluble structure of disulfide bond, electron density is high, the electrophilicity that has and with
The ability of radical reaction, therefore with significant inoxidizability.Clinically cause for treating diabete peripheral herve pathology
Cacesthesia.In the intervention and treatment of diabetic nephropathy, Zhou Jianling in 2015 reports lipoic acid to early diabetes kidney
Sick clinical efficacy influence (《Chinese practical medicine》, the phase of volume 10 6, page 143 in 2015), controlled with patient with melbine
Lipoic acid capsule is given while treatment, successive administration 28 days can reduce the effect of Urinary Microalbumin Excretion rate, to treatment early stage sugar
The sick nephrosis effect of urine is notable.Wang Hui etc. reported influence research of the lipoic acid to diabetic nephropathy fibrosis in 2011
(《Liaoning Medical University's journal》, the phase of volume 32 6, page 489 in 2011), lipoic acid continuous gavage is administered 12 weeks, as a result shows that sulphur is pungent
Acid can play the effect of its anti-fibrosis by suppressing the expression of the grade of nephridial tissue TGF-β 1, so as to delay STZ diabetes rats
The development of renal fibrosis.However, because lipoic acid is not enough protruded the intervention and protective effect of diabetic nephropathy, because
This important drugs also not turned into so far during diabetic nephropathy is treated is used.Current lipoic acid is to diabetic nephropathy
Focused mostly in Intervention Mechanism research in Antioxidation Mechanism, whether such medicine also has the other mechanism of action for intervening diabetic nephropathy
Then study less.
Lipoamide (Lipoamide), is a kind of derivative of alpha-lipoic acid.
Lipoamide can remove interior free yl, promote the generation of endogenic antioxidant, because it is efficient anti-oxidant
Effect causes the interest of researcher.The research such as Li finds that lipoamide can improve mitochondria work(by suppressing oxidative stress
Retinal pigment epithelium (volume 44, page 1465 of Free Radic Biol Med.2008) can be protected.The research hair such as Zhao
Existing lipoamide can play indirect antioxidant and act on (PLoS by activating Mitochondrial regeneration and II phase antioxidase
One.2015 volumes 10, e0128502).Lipoamide through listing, but there is not yet sulphur abroad as Liver protection medication
Report of the caprylamide in terms of delaying and treating the drug effect of diabetic nephropathy, also has no its study on mechanism.
The content of the invention
It is an object of the invention to provide purposes of the lipoamide in delaying and treating diabetic nephropathy.
The lipoamide of the present invention is for preparing the purposes in delaying and treating the composition of diabetic nephropathy.
The lipoamide of the present invention is improving the combination of kidney fibrosis lesion, improvement Renal Function in Rats and metabolism for preparing
Purposes in thing.
The lipoamide of the present invention is consolidated for preparing reduction diabetes rat kidney weight/weight ratio, 24h Urine proteins, total courage
Purposes in alcohol, the composition of triglycerides.
The lipoamide of the present invention is suppressing the effect of kidney of diabetic rats oxidative stress for preparing, and recovers kidney group
Knit TAC (Total antioxidant capacity, T-AOC), total number born (Total
Superoxide Dismutase, T-SOD), the activity of catalase (Catalase, CAT), and significantly reduce nephridial tissue third
Purposes in the composition of dialdehyde (Malondialdehyde, MDA) level.
The lipoamide of the present invention is played for preparing the expression of reduction renal interstitial collagen I (Collagen I)
Anti-diabetic renal fibrosis is acted on, hence it is evident that improve the purposes in the composition of kidney fibrosis lesion.
The lipoamide of the present invention is promoting the consideration convey record co-suppression factor S noN of kidney fibrosis key regulatory for preparing
Purposes in the composition of the expression of albumen.
The lipoamide of the present invention is reducing the consideration convey record co-suppression factor S noN of kidney fibrosis key regulatory for preparing
Purposes in proteins ubiquitin level, the composition of raising SnoN protein stabilities.
The optical antipode of lipoamide, hydrate, hydrogenation addition product or the precursor of the present invention, with lipoamide phase
Same or close improves kidney fibrosis lesion, improves the function of Renal Function in Rats and metabolism, to delay and treat diabetes kidney
Complication.
Described " hydrogenation addition product " refers to the sulfhydryl compound of the hydrogenated ring-opened formation of disulfide bond in structure formula (I)；
Described " precursor " refer to be metabolized or chemically reacted in mammal body can be transformed into structure formula (I) a kind of compound or
Its analog.
The composition of the present invention includes pharmaceutical composition and dietary supplement (such as Halth-care composition), as long as they contain
Lipoamide is used as improvement kidney fibrosis lesion, improvement renal function and the active component of metabolism.
Lipoamide of the invention or its optical antipode, hydrate, hydrogenation addition product or precursor are used to prepare and can improved
Purposes in the composition of kidney fibrosis lesion, improvement Renal Function in Rats and metabolism.
SD rat models of the invention by setting up diabetic nephropathy, using lipoic acid as positive control, by lipoamide
Rat is given with lipoic acid gavage.As a result show that lipoamide can substantially reduce diabetes rat kidney weight/weight ratio (KW/ after 6 weeks
BW), blood T-CHOL (TC), triglycerides (TG), 24h Urine proteins (24h UP)；T-AOC, T-SOD, CAT activity are significantly higher than
DM groups, and MDA levels are significantly reduced, and the T-AOC and T-SOD of lipoamide treatment group activity recover more aobvious compared with lipoic acid group
Write, and the reduction of MDA levels also becomes apparent from than lipoic acid group；H＆E, Masson coloration result show lipoamide group kidney fibrosis disease
Become apparent improvement, and the expression of lipoamide group renal interstitial collagen I (collagen I) is significantly lower than lipoic acid group；Sulphur is pungent
The expression increase of acid amides treatment group SnoN protein levels；The SnoN albumen of ubiquitination is reduced, the stability increase of SnoN albumen,
And find that the expression and activation of transforming growth factor β activation kinases (TAK-1) are reduced.The above results show that lipoamide can change
Kind Renal Function in Rats and metabolism, and with the effect of increase SnoN protein expression levels；Simultaneously can also be by suppressing TAK1 table
Reach and activate, improve the stability of SnoN albumen, and then reduce Collagen I expression and play anti-diabetic kidney fibrous
Change effect.
Embodiment
The experimental method of unreceipted actual conditions in following experiments, generally implements according to the normal condition of this area.
1. material
Healthy cleaning grade male Sprague-Dawley (SD) rat, body weight (180 ± 20) g is biological by magnificent Fukang, Beijing
Science and Technology Co., Ltd. provides, and lot number is SCXK (capital) 2009-0004.Steady extraordinarily type blood glucose meter (Johson ＆ Johnson), it is ultralow
Temperature refrigerator (Sanyo), high speed low temperature centrifugal machine (Beckman), Bayer1650 automatic clinical chemistry analyzers (Beckman), electrophoresis
System and electrotransfer device (Amersham), gel imaging system (Bio-Rad).Positive control drug lipoic acid obtains for purchase
(Xi'an Ze Bang bio tech ltd)；Test-compound lipoamide is synthesized according to following method.
The preparation method of lipoamide：Lipoic acid 30.0g and 300mL dichloromethane is added in round-bottomed flask, is stirred molten
I-hydroxybenzotriazole 20.8g is added after solution at room temperature, finishes after stirring 0.5h, dicyclohexylcarbodiimide is added in three batches
28g, adds and stopping reaction after 10h is stirred at room temperature, and reaction solution is washed with water three times (150mL/ times) and divides liquid to concentrate afterwards, obtains yellow and consolidates
40 grams of body.Solid 15.0g is taken to be dissolved in 90mL tetrahydrofurans, stirring is lower to add 40mL concentrated ammonia liquors, is stirred at room temperature after 8h and subtracts
Pressure is evaporated off in organic solvent, residue adding 50mL water, is stirred at room temperature after 2h, suction filtration, 40 DEG C of filter cake is dried under reduced pressure, and obtains sulphur decoyl
Amine crude product.It is pale yellow powder shape solid, yield 84.7% that crude product obtains lipoamide with ethyl alcohol recrystallization afterwards twice.Structural characterization
Data：1H NMR (d-DMSO, 400MHz), δ:7.21(s,1H)；6.68(s,1H)；3.58-3.54(m,1H)；3.14-3.06
(m,1H)；2.77(s,1H)；2.46-2.36(m,1H)；2.03-1.98(m,2H)；1.81-1.62(m,1H)；1.56-1.28
(m,6H)。13C NMR (d-DMSO, 100MHz), δ:174.66,56.69,40.05,38.63,35.46,34.70,28.92,
25.40.ESI-MS：m/z:228.1[M+Na]+。
2. method
2.1 animal model replications and packet：After SD rats adaptability is fed one week, tail vein injection is dissolved in pH4.5's
The chain assistant urea rhzomorph (dosage is 55mg/kg) of the sterile citric acid-sodium citrate buffer solutions of 0.01mol/L builds diabetes rat
Model.Rat fasting blood-glucose, blood glucose >=16.7mmol/L and the positive then modeling success of glucose in urine are surveyed after 72h.Rat is randomly divided into sugar
The sick group (DM groups) of urine, lipoic acid treats group (ALA groups) and lipoamide treatment group (ALM groups).Into after mould 2 weeks, gavage side is taken
Formula gives lipoic acid and lipoamide treatment, lipoic acid and lipoamide respectively it is muddy be suspended from 5% sodium carboxymethylcellulose (CMC-
Na) in the aqueous solution, given low is 150mg/kg/d, weekly administration 6 days；And set mouse age identical Normal group (NC groups),
The CMC-Na aqueous solution of gavage comparable sodium Isodose.All rats give fed standard chow, and free water monitors weekly one
Secondary blood glucose and body weight.All rats are put to death after 6 weeks.
Experimental data is represented with mean ± standard deviation (x ± s), using SPSS17.0 softwares, compares neat using variance between group
Property examine, make one-way analysis of variance, P<Represented when 0.05 statistically significant.
2.2 specimen collection：Rat is put to death first 1 day and collects 24h urine with metabolic cage, records urine volume, takes part urine centrifugation
- 20 DEG C of preservations afterwards；Fasting 6-8h, weighs after etherization before putting to death, femoral artery puncture blood sampling, -20 DEG C of preservations of separation serum；
Open abdomen and take bilateral renal, remove coating and peripheral adipose tissue, record kidney weight/weight ratio (KW/BW) of weighing, respectively with more than 4%
Polyformaldehyde is fixed and -80 DEG C of preservations.
Experimental example 1：Lipoic acid is with lipoamide to Rat renal weight/weight ratio, serum total cholesterol (TC), triglycerides
(TG), the contrast experiment of the influence of Urine proteins
Enzyme assay detection serum total cholesterol (TC) and triglycerides (TG)；Pyrogallol red colorimetric method surveys Urine proteins
(urine Protein, UP), is operated by kit specification, and urinary protein concentrations are 24h urine albumen amounts (24h with urine volume product
UP)。
Table 1 test 6 weeks after each group Rat renal weight/weight ratio, 24h Urine proteins, T-CHOL, triglycerides change (x ±
s)
Compared with normal group,*P<0.05；Compared with diabetes group,#P<0.05；Compared with lipoic acid group,ΔP<0.05
From table 1, KW/BW, TC, TG and 24h UP of lipoamide group (ALM) and lipoic acid group (ALA) reduce (P
<, and lipoamide is significantly stronger than lipoic acid to TC, TG reduction effect 0.05).
Experimental example 2：Lipoic acid or lipoamide are to rat TAC (T-AOC), catalase (CAT), super
The influence of superoxide dismutase (T-SOD), MDA (MDA)
All indexs are determined using colorimetric method with reference to kit specification.
The comparison (x ± s) of each group rat oxidative stress after table 2 is tested six weeks
Compared with normal group,*P<0.05；Compared with diabetes group,#P<0.05；Compared with lipoic acid group,ΔP<0.05
From table 2, diabetes rats kidney T-AOC, T-SOD and CAT activity reduce (P compared with normal group<
0.05), lipoic acid and lipoamide can raise DM rat kidney T-AOC, T-SOD and CAT activity (P<, and sulphur is pungent 0.05)
Acid amides up-regulation Renal of Diabetic Rats T-AOC and T-SOD activity are compared with lipoic acid significantly (P<0.05)；And diabetes rats
Kidney MDA contents are compared with normal group compared to rise (P<0.05), lipoic acid and lipoamide can lower Renal of Diabetic Rats MDA
Content, especially becomes apparent from (P with the effect of lipoamide<0.05), illustrate that lipoic acid and lipoamide are respectively provided with anti-oxidation stress
Effect, so as to mitigate or delay diabetic nephropathy to develop, and the antioxidation of lipoamide is significantly stronger than sulphur
Octanoic acid.
Experimental example 3：The influence of lipoic acid (ALA) or lipoamide (ALM) to renal tissues of rats Morphology
Paraformaldehyde fixes nephridial tissue, and the paraffin section of 3 μ m-thicks, row H＆E and Masson dyeing, om observation kidney group is made
Knit Morphology.The infringement method that tubulo-interstital is assessed in H＆E dyeing is as follows：Randomly selected in cortex renis under 10 20 times of mirrors
The visual field carries out system evaluation (1-6 grade).Injury score, which sentences grade scale, includes tubular ectasia, and renal tubule atrophy, cast is formed,
The scope of interstitial monocyte and extrtacellular matrix deposition accounts for the percentage of total visual field, and (1=is less than 10%；2=10-25%；3=
26-50%, 4=51-75%, 5=76-95%, more than 6=95%).
H＆E and Masson dyes visible normal rat renal tubule clear in structure, renal cells marshalling, substrate
Film is complete, and interstitial has no inflammatory cell infiltration；The expansion of DM group Rat renals lumen is obvious, renal tubular basement membrane irregular thickening,
There is empty balloon-shaped change in renal cells, and inflammatory cell infiltration, renal tubular interstitium Masson stained positive things occurs in interstitial
Matter increases；ALA and ALM group rat kidney lesions have different degrees of improvement, and renal tubular interstitium Masson stained positive materials are bright
Aobvious to reduce, inflammatory cell infiltration mitigates.System injury grading ALM groups are less than ALA groups after H＆E dyeing, it is seen that lipoamide is to kidney
The injury protection effect of tissue is better than lipoic acid.
Experimental example 4：The influence of lipoic acid (ALA) and lipoamide (ALM) to rat Collagen I
Using immunohistochemistry staining method, SP two-step methods detect Collagen I in the distribution of each group renal tissues of rats and
Expression.Paraffin section de-waxing aquation, after repairing antigen and 5%BSA closings through pancreatin, Collagen I (1: 100), 4 DEG C of incubations
Biotinylation secondary antibody is added overnight, 30min is incubated at room temperature, and SP reagent 3- amino -9- ethyl card azoles (3-amino-9- is added dropwise
Ethylcarbozole, AEC) develop the color, haematoxylin is redyed, and PBS replaces primary antibody, makees negative control.Collagen I's is heavy
Product is analyzed using quantitative iamge analysis system (Image-Pro+v 6.0)：Randomly selected in cortex renis under 10 40 times of mirrors
The visual field containing glomerulus is analyzed, and determines the region of positive staining, and lumen of artery is excluded outside research, calculates sun
Property dyeing area account for the ratio of the gross area.
As a result show that normal rat nephridial tissue Collagen I positive stainings are primarily present around blood vessel and cytoplasm, and
In its positive staining showed increased of diabetes rat, it is seen that renal tubular basement membrane strong positive is dyed.Lipoic acid and lipoamide are controlled
Collagen I expression is significantly reduced after treatment, and Collagen I expression is compared with lipoic acid especially after lipoamide treatment
Group reduction becomes apparent from (P<0.05), show that lipoic acid and lipoamide can play anti-sugar by reducing Collagen I expression
The sick renal fibrosis effect of urine, and the effect of lipoamide is stronger compared with lipoic acid.
Experimental example 5：Lipoamide (ALM) is to the expression of renal tissues of rats transforminggrowthfactor-β1, TAK1 protein expressions and work
The influence of change
The each group renal cortex of rats of -80 DEG C of preservations is taken, every sample takes 200mg, is separately added into after histone extract solution
Centrifuging and taking supernatant is homogenized, each group protein concentration is determined with BCA kits (the green skies), is calculated by measured concentration per swimming lane institute
Volume is needed, sample loading buffer is added and boils 10min, be separated by electrophoresis, transferred on pvdf membrane through 8% PAGE gel,
5% skimmed milk power room temperature closes 1h, is separately added into β-actin, TGF-β 1, TAK1, p-TAK1 primary antibody, working concentration is respectively 1:
4000、1:300、1:1000、1:1000,4 DEG C of incubation 12-24h；Next day, TBST is washed after film, adds corresponding horseradish peroxidase
Secondary antibody (concentration is 1: 4000) the incubation at room temperature 1h of enzyme mark, plus ECL fluorescence developing liquid, gel imager exposure, image
Each band adjustment bulking value of lab software analysis, each sample repeats 3 times using β-actin protein bands as internal reference, as a result
Represented with target protein/β-actin values.
As a result show, TGF-β 1 and TAK1, P-TAK1 expression are less in Normal group (NC)；And diabetes group (DM)
Middle TGF-β 1 and TAK1, P-TAK1 dramatically increase (P compared with normal group<0.05)；Lipoamide (ALM) treatment after TGF-β 1 and
TAK1, P-TAK1 significantly reduce (P compared with DM groups<0.05).Show that lipoamide can suppress TGF-β 1 and express, suppress simultaneously
TAK1 expression and activation.
Experimental example 6：The influence of lipoamide (ALM) new albumen N (SnoN) protein expression level related to rat Ski
The detection of SnoN protein levels：The each group renal cortex of rats of -80 DEG C of preservations is taken, every sample takes 200mg, added respectively
Enter after histone extract solution homogenate centrifuging and taking supernatant, each group protein concentration is determined with BCA kits (the green skies), by being surveyed
Obtain concentration and calculate volume needed for per swimming lane, add sample loading buffer and boil 10min, be separated by electrophoresis through 8% PAGE gel,
Transfer on pvdf membrane, 5% skimmed milk power room temperature closing 1h is separately added into β-actin, SnoN primary antibodies, working concentration difference
For 1:4000、1:300,4 DEG C of incubation 12-24h；Next day, TBST is washed after film, adds corresponding horseradish peroxidase-labeled
Secondary antibody (concentration is 1: 4000) is incubated at room temperature 1h, plus ECL fluorescence developing liquid, gel imager exposure, image lab softwares point
Each band adjustment bulking value is analysed, each sample is repeated 3 times and target egg is used using β-actin protein bands as internal reference, as a result
In vain/β-actin values are represented.In Normal group, SnoN height expression；SnoN expressions are aobvious compared with normal group in diabetes group (DM)
Write and lower (P<0.05)；SnoN expressions substantially up-regulation (P compared with diabetes group in lipoamide group<0.05),.Show
Lipoamide can recover the key regulatory Protein S noN of kidney fibrosis expression.
Experimental example 7：The influence of lipoamide (ALM) new albumen N (SnoN) ubiquitination level related to rat Ski
The detection of SnoN ubiquitination levels：The each group renal cortex of rats of -80 DEG C of preservations is taken, every sample takes 200mg, respectively
Add and centrifuging and taking supernatant is homogenized after histone extract solution, each group protein concentration is determined with BCA kits (the green skies).Pass through
Immunoprecipitation (Immunoprecipitation, IP) isolates and purifies SnoN, and the SnoN of extraction is added into sample loading buffer boils
10min, is separated by electrophoresis through 8% PAGE gel, transfers on pvdf membrane, 5% skimmed milk power room temperature closing 1h, respectively
Ubiquitin, SnoN are added, IgG primary antibodies, working concentration is respectively 1:1000、1:300,4 DEG C of incubation 12-24h；Next day, TBST washes film
Afterwards, secondary antibody (concentration is 1: 4000) the incubation at room temperature 1h of corresponding horseradish peroxidase-labeled, plus ECL fluorescence developings are added
Liquid, gel imager exposure compares Blot results and can detect SnoN proteins ubiquitin levels.In Normal group (NC), ubiquitin
The SnoN protein expressions of change are less；And the SnoN albumen of ubiquitination is dramatically increased compared with normal group in diabetes group (DM)；Sulphur decoyl
The SnoN albumen of ubiquitination is significantly reduced compared with diabetes group in amine group (ALM).Show that lipoamide can reduce SnoN eggs
White ubiquitination degraded, increases the stability of SnoN albumen.
The above described is only a preferred embodiment of the present invention, not making any formal limitation to the present invention, appoint
What is without departing from technical solution of the present invention content, and what the technical spirit according to the present invention was made to above example any simply repaiies
Change, equivalent variations and modification, in the range of still falling within technical solution of the present invention.
Claims (8)
1. lipoamide is for preparing the purposes in delaying and treating the composition of diabetic nephropathy.
2. lipoamide is for preparing the use in the composition for improving kidney fibrosis lesion, improvement Renal Function in Rats and metabolism
On the way.
3. lipoamide is for preparing reduction diabetes rat kidney weight/weight ratio, 24h Urine proteins, T-CHOL, triglycerides
Composition in purposes.
4. lipoamide is suppressing the effect of kidney of diabetic rats oxidative stress for preparing, recover nephridial tissue total antioxidation
Ability（Total antioxidant capacity, T-AOC）, total number born (Total Superoxide
Dismutase, T-SOD), the activity of catalase (Catalase, CAT), and significantly reduce nephridial tissue MDA
Purposes in the composition of (Malondialdehyde, MDA) level.
5. lipoamide plays anti-diabetic kidney reducing the expression of renal interstitial collagen I (Collagen I) for preparing
Dirty Fibrosis parameters, hence it is evident that improve the purposes in the composition of kidney fibrosis lesion.
6. lipoamide is in the expression for the consideration convey record co-suppression factor S noN albumen for promoting kidney fibrosis key regulatory for preparing
Purposes in the composition of level.
7. lipoamide records co-suppression factor S noN proteins ubiquitins in the consideration convey that kidney fibrosis key regulatory is reduced for preparing
Purposes in level, the composition of raising SnoN protein stabilities.
8. the optical antipode of lipoamide, hydrate, hydrogenation addition product or precursor, with identical or close with lipoamide
Improve kidney fibrosis lesion, improve the function of Renal Function in Rats and metabolism, to delay and treat the group of diabetic nephropathy
Purposes in compound;
Described hydrogenation addition product refers to the sulfhydryl compound of the hydrogenated ring-opened formation of disulfide bond in structure formula (I)；
Described " precursor ", which refers to be metabolized or chemically reacted in mammal body, can be transformed into one kindization of structure formula (I)
Compound or its analog.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710249412.2A CN107028938B (en) | 2017-04-17 | 2017-04-17 | Application of lipoamide in preparing composition for delaying and treating diabetic nephropathy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710249412.2A CN107028938B (en) | 2017-04-17 | 2017-04-17 | Application of lipoamide in preparing composition for delaying and treating diabetic nephropathy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107028938A true CN107028938A (en) | 2017-08-11 |
| CN107028938B CN107028938B (en) | 2020-09-15 |
Family
ID=59536442
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710249412.2A Active CN107028938B (en) | 2017-04-17 | 2017-04-17 | Application of lipoamide in preparing composition for delaying and treating diabetic nephropathy |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107028938B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101396357A (en) * | 2007-09-29 | 2009-04-01 | 中国科学院上海生命科学研究院 | Use of lipoamide in preventing and treating insulin resistance |
| CN101787013A (en) * | 2009-01-22 | 2010-07-28 | 杭州民生药业有限公司 | Novel lipoamide derivatives and preparation method and pharmaceutical application thereof |
-
2017
- 2017-04-17 CN CN201710249412.2A patent/CN107028938B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101396357A (en) * | 2007-09-29 | 2009-04-01 | 中国科学院上海生命科学研究院 | Use of lipoamide in preventing and treating insulin resistance |
| CN101787013A (en) * | 2009-01-22 | 2010-07-28 | 杭州民生药业有限公司 | Novel lipoamide derivatives and preparation method and pharmaceutical application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 云鹏等: "α－硫辛酸联合氯沙坦治疗早期糖尿病肾病的临床观察", 《中国医院药学杂志》 * |
| 林海燕等: "氧化应激与糖尿病肾病关系的研究进展", 《医学综述》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107028938B (en) | 2020-09-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ajagbonna et al. | Haematological and biochemical changes in rats given extract of Calotropis procera | |
| CN104080454B (en) | Methods of inhibiting muscle atrophy | |
| Kirby | Cataracts produced by triparanol.(MER-29). | |
| CN106994131B (en) | A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy | |
| CN107441078B (en) | A kind of pharmaceutical composition and its preparation method and application for treating diabetes | |
| RU2541093C2 (en) | Using 5?-androstane-3(,5,6(-triol for preparing neuroprotective drugs | |
| Moon et al. | Ninety-day oral toxicity study of rice-derived γ-oryzanol in Sprague-Dawley rats | |
| Bae et al. | Effect and putative mechanism of action of ginseng on the formation of glycated hemoglobin in vitro | |
| McClean et al. | 17α-Estradiol is neuroprotective in male and female rats in a model of early brain injury | |
| Alvarado-Vásquez et al. | Oral glycine administration attenuates diabetic complications in streptozotocin-induced diabetic rats | |
| CN108853080A (en) | Application of the procyanidine in the functional food or drug that preparation improves obese males sperm quality | |
| CN115400116A (en) | Application of luteolin and derivatives thereof in medicines for regulating uric acid secretory protein and kidney injury factor expression level | |
| CN107028938A (en) | Application of the lipoamide in delaying and treating diabetic nephropathy | |
| Nabi et al. | Role of biometals in activation of immune cum inflammatory response in ovine ageing eye: a potential model for understanding human geriatric eye diseases | |
| Guo et al. | Therapeutic effect of dexmedetomidine on myocardial ischemia reperfusion injury in type 2 diabetic rat model under P13k/akt pathway | |
| CN106667992B (en) | Use of coniferyl aldehyde or coniferyl aldehyde and vanadium compound composition for preventing and treating neurodegenerative diseases | |
| CN112206231A (en) | Application of matrine derivative in treating non-alcoholic steatohepatitis | |
| CN101703503A (en) | Application of organic dicarboxylic acid salts in preparing medicaments for preventing and treating cataract | |
| JP6266666B2 (en) | Pharmaceutical composition and method for treating ocular diseases associated with angiogenesis | |
| CN108888652A (en) | A kind of renal lobe beggarweed alcohol extract and its application | |
| JP7353246B2 (en) | Nucleic acids and their use for the treatment or improvement of male metabolic syndrome | |
| CN108310026A (en) | Application of the evening primrose oil in terms of improving Stein-Leventhal syndrome chronic inflammation | |
| CN104473915B (en) | Sivelestat is as the application for the treatment of epilepsy medicament | |
| CN111973606B (en) | New use of compound for treating vascular proliferation and diabetic retinopathy | |
| CN113476607B (en) | SLC12A5 and application of inhibitor thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20191225 Address after: 201109 No. 3599 Yuanjiang Road, Minhang District, Shanghai Applicant after: SHANGHAI CIRUI MEDICINE TECHNOLOGY CO.,LTD. Address before: 550004 No.4, Beijing Road, Guiyang City, Guiyang City, Guizhou Province Applicant before: GUIZHOU MEDICAL UNIVERSITY |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230404 Address after: 550004 No. 4, Beijing Road, Guiyang, Guizhou Patentee after: GUIZHOU MEDICAL University Address before: 201109 No. 3599 Yuanjiang Road, Minhang District, Shanghai Patentee before: SHANGHAI CIRUI MEDICINE TECHNOLOGY CO.,LTD. |
|
| TR01 | Transfer of patent right |