CN103948581A - Application of combination of levocarnitine and L-arginine in preparation of drugs for treatment of diabetic retinopathy nerve damage - Google Patents
Application of combination of levocarnitine and L-arginine in preparation of drugs for treatment of diabetic retinopathy nerve damage Download PDFInfo
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Abstract
The invention discloses application of a pharmaceutical composition containing L-arginine and levocarnitine in combined preparation of drugs for treatment of diabetic retinopathy nerve damage, and particularly relates to application of the pharmaceutical composition in preparation of the drugs for treatment of retinal nerve damage and Muller cell injury of diabetic retinopathy nerve damage. Cell and animal experiments show that compared with the single use of the levocarnitine, the diabetic retinopathy nerve damage and particularly the retinal nerve damage and Muller cell injury of diabetic retinopathy nerve damage can be more effectively protected by combination of the levocarnitine and the L-arginine, and the technical effect of synergistic interaction can be achieved.
Description
Technical field
The present invention relates to levocarnitine associating L-arginine and become the application in nerve injury medicine in preparation treatment of diabetic retinopathy.
Background technology
Along with human living standard's generally raising and the prolongation of the average life span, the sickness rate of diabetes significantly raises both at home and abroad at present.Diabetes have many complication, wherein diabetic renal papillary necrosis (retinal retinopathy, DR) be one of its common, the most serious microvascular complication at eye, one of main blinding oculopathy, be mainly that its sickness rate and the order of severity increase resistance to along with the prolongation of diabetic duration because thereby retinal microvascular generation pathological changes causes a series of pathological change of retina.If studies have reported that diabetic duration as long as five years, DR sickness rate can reach 44.4%, and as diabetic duration reaches 7 years, the sickness rate of DR can be up to 56%.
Think that at present basic pathology of DR changes as follows: 1. the selectivity of pericyte is lost; 2. thickening of basement membrane; 3. the formation of microangioma; 4. the hypertrophy of endotheliocyte; 5. the formation of new vessels.Wherein the selectivity of pericyte is lost the pathological change that is regarded as its earliest period.In normal retina blood capillary, pericyte is about 1: 1 with the ratio of the quantity of endotheliocyte, and in diabetes and complication patient thereof, both ratios are about 1: 4, even lower.In recent years research shows, the generation of diabetic renal papillary necrosis is relevant with oxidative stress.Response to oxidative stress refers to that chemical factor and environmental factors cause that interior free yl increases, and causes antioxidase activity to reduce, DNA damage, thus promote tumor growth, cardiovascular disease and apoptosis.At eye, by irritation cell apoptosis, destroy the integrity of cell membrane, cause amphiblestroid irreversible damage.Hyperglycemia strengthens the aerobic oxidation of glucose, shows as that the electronics producing in tricarboxylic acid cycle process increases, mitochondrial proton gradient increases, and then the electronics spilling and O
2in conjunction with generating O
2 -, active oxygen (ROS) produces increases, and causes response to oxidative stress.The researchs such as Takeshi Nishikawa show: hyperglycemia can increase the generation of mitochondrion ROS, and this process is relevant with the generation of diabetic angiopathy.ROS should attack insatiable hunger profit fatty acid, and retina is the membrane tissue that is rich in polyunsaturated fatty acid, and compared to its hetero-organization, the oxidability of the picked-up ability profit glucose to oxygen is stronger.So when diabetes pathological changes, retina is easily subject to the damage of oxidative stress.In addition, hyperglycemia can cause that many metabolic pathway reactions of body change, comprise protein non-enzyme glycosylation end-product (AGEs) formation, protein kinase C activation, polyhydric alcohol Developmental and Metabolic Disorder, amidohexose pathway activation, also impel the increasing expression of various inflammatory factors and cell adhesion factor etc. simultaneously.Above-mentioned a series of pathophysiological change causes the death of retinal blood tube wall pericyte, basement membrane thickened, makes blood vessel wall insufficiency, has destroyed blood-retina barrier function, and then has caused osmotic pressure to increase, cellular edema.
Neurogliocyte (neuroglial cell) claim again glial cell, glial cell, is one of neural composition unit.Recent study shows, retina M ü ller cell is topmost glial cell in retina, also be a kind of neurogliocyte of particularization, there is the normal retinal structure of maintaining, participate in forming blood-retina barrier, support and trophic nerve unit, regulate the physiological functions such as neurotransmitter circulation.It plays very important effect in the multiple pathological process of retinopathy.
Due to the pathogenesis complexity of diabetic retinopathy, all lack effective treatment means all the time.Laser and operative treatment can not stop the PD of DR.Along with to the pathogenetic further investigation of DR, it is found that Drug therapy can block multiple approach of DR morbidity.But at present conventional medicine, as anti-vascular endothelial growth factor and glucocorticoid etc., has dispute at aspects such as evaluating its treatment effectiveness and Long-Term Safety.Therefore, find medicine safely and effectively, for the early treatment of DR, particularly important to reducing blind rate.
Levocarnitine (L-carnitine, LC) have another name called L-carnitine, it is the necessary natural materials of mammal energy i (in vivo) metabolism, mainly in the tissues such as liver, kidney, heart, change into gamma-butyrobetaine by trimethyl lysine, convert it into carnitine and synthesize by liver, kidney and cerebral tissue again, participating in lipid metabolism in body, body cell energy metabolism generation and transhipment are played an important role, is one of main source of the histoorgan metabolism institute energy requirements such as the heart, brain, kidney.LC also has the functions such as the protein degradation of promotion, antioxidation, Cell protection film simultaneously.Levocarnitine has been widely used in the treatment of the diseases such as cardiomyopathy, hepatic disease, dialysis patient, diabetes, male sterility, neuromuscular disease, kidney disease at present.Under normal circumstances, LC can, by long-chain fatty acid by being arranged in the carnitine acyl transferring enzyme-I of mitochondrial outer membrane and the carnitine acyl transferring enzyme-II of inner membrance is transported to mitochondrion, then can carry out beta oxidation by acyl coenzyme A (CoA) in the effect of cyclophorase in mitochondrion.Lipid metabolism generation obstacle when ischemia, anoxia, can cause long-chain acyl carnitine in mitochondrion, to be piled up and the accumulation of acyl-CoA, cause free carnitine to be reduced by a large amount of consumption, and the acyl-CoA piling up can cause the change of membrane structure, the disintegrate of film phase to cause cell death.As there being enough free carnitines in body, the acyl-CoA that can make to pile up enters and in mitochondrion, under the effect of cyclophorase, carries out beta oxidation, thereby it is unbalance to correct energy due to lipid metabolism obstacle, anti-upper lipid peroxidation.There are some researches show in addition; LC is except promoting fatty acid to be oxidized utilization by beta oxidation; it also can be used as a kind of oxygen free radical scavenger and effectively removes the oxygen-derived free radicals in body, its at enhancing body oxidative stress, prevent that the aspects such as lipid peroxidation have obvious protective effect.
Chen Jie (Master's thesis " levocarnitine is cultivated protective effect and the Mechanism Discussion of Rat retina neurocyte to external high sugar "), Yin Xiaolin (Master's thesis " protective effect of levocarnitine to retinal neuronal cell under the sugared condition of height "), the Yu Changhong (experimentation of levocarnitine to the protective effect of diabetic retinal tissue in rat ganglionic cell; " Chinese Pharmacological Bulletin ", the 29th volume o. 11th in 2013) all with regard to levocarnitine, the protective effect of retinal neuronal cell is studied.These researchs show, the protective effect of the retinal neuronal cell oxidative damage that levocarnitine causes high sugar is relevant with its performance anti-oxidation characteristics.As a kind of antioxidant, levocarnitine can reduce oxygen-derived free radicals generation, strengthen the oxidation resistance of cell, and can stability line mitochondrial membrane potential, improve mitochondrial function, thereby suppress the apoptosis that under high sugared state, oxidative stress causes.
In addition, Zhang Ying (Master's thesis " levocarnitine is cultivated protective effect and the mechanism of Rat retina M ü ller cell to external high sugar ") has studied the protective effect of levocarnitine to retina M ü ller cell.Research shows; high sugar is mainly because ROS rolls up the oxidative stress damage causing to the damage of retina M ü ller cell; and LC has protective effect to high sugar damage retina M ü ller cell; can suppress the apoptosis of M ü ller cell; its mechanism may generate with minimizing ROS; suppress mitochondrial membrane permeability, maintain mitochondrial membrane potential relevant.
But in view of the complicated pathogenesis of diabetic retinopathy, and multiple approach of morbidity, how further looking for medicine is more safely and effectively the emphasis that people pay close attention to.
Summary of the invention
In view of the deficiencies in the prior art; the inventor finds by theoretical and a large amount of cells of combination, zoopery; levocarnitine associating L-arginine can coordinating protection diabetic renal papillary necrosis nerve injury, especially diabetic renal papillary necrosis retina neural damage and M ü ller cell injury.
Therefore, the object of this invention is to provide a kind of pharmaceutical composition, it is characterized in that, in described compositions, contain levocarnitine and L-arginine, be used for the treatment of diabetic renal papillary necrosis nerve injury.
Further, the mol ratio of described levocarnitine and described L-arginine is 1: 1～10.
Further, the dosage form of described compositions is tablet, capsule, powder, granule, solution, Emulsion or suspensoid.
Another object of the present invention is to provide the application in the retina neural damage of levocarnitine associating the L-arginine application in the medicine of preparation treatment of diabetic retinopathy change nerve injury, especially treatment of diabetic retinopathy change and the medicine of M ü ller cell injury.
Further, in above-mentioned application, the mol ratio of levocarnitine and L-arginine is 1: 1～10, be further preferably 1: 3～and 8, most preferably be 1: 6.
Further, in above-mentioned application, levocarnitine and L-arginine can carry out administration with regular dosage forms such as tablet, capsule, powder, granule, solution, Emulsion, suspensoids.
L-arginine (L-Arginine, L-Arg) is a kind of alpha amino acid, and it is all playing the part of important role in multiple processes, and such as cell division, wound restore, discharge ammonia, immunologic function and secreting hormone etc.L-arginine and amino molecule interact and generate NO and L-Citruline under the catalysis of nitricoxide synthase (nitric oxide synthase, NOS).NO can activate guanylate cyclase (cGMPase) and generate cyclic guanosine monophosphate (cGMP), and then brings into play its biological effect, and this approach is called NO approach.NOS is a kind of monoamine oxidase, MAO of iron content, it is the rate-limiting enzyme in NO building-up process, according to the difference of tissue distribution scope, Ca-dependent and the gene expression of NOS, NOS is divided into induced NOS (inducible NOS, iNOS), neuron NOS (neuronal NOS, and endothelium in type NOS (endothe NOS, eNOS) nNOS).The main biological action of NO is: the NO of (1) physiological dose can participate in the information transmission in immunoreation and inflammatory process, and large agent NO can cause neuronal cell injury or kill normal cell, and causing cell to ask Information Conduction obstacle.(2) information transfering action: NO is the one neurotransmitter that is not true to type that in autonomic nervous system, neuron discharges, be present in widely each internal organs in human body, mode with local diffusion acts on adjacent cells, the cGMP performance biological effect generating by NO approach: (3) vasodilator: NO is combined with GC and is activated sGC, thereby cGMP level is raise and bring into play its biological effect: by loose vascular smooth muscle, vasodilator regulates peripheral vascular resistance, and participates in regulation and control cerebral circulation, coronary artery circulation and pulmonary circulation; (4) inflammatory mediator: interleukin I (IL-1), tumor necrosis factor (TNF), lipopolysaccharide can be induced the expression of neutrophilic granulocyte and macrophage L-Arg-NO approach, generate a large amount of NO, can kill intracellular bacteria, tumor cell and parasite on the one hand, can damage again on the other hand normal structure, cause cell death, participate in the autoimmune response of body.
Beneficial effect of the present invention is: prove by cell and zoopery; compared with independent use levocarnitine; associating L-arginine can be protected diabetic renal papillary necrosis nerve injury more effectively; especially the retina neural of diabetic renal papillary necrosis damages and M ü ller cell injury, has reached the technique effect of Synergistic.
Detailed description of the invention
The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not subject to any restriction of specific embodiment, but be limited by claim.
Experimental example 1
1.1 laboratory animal
The standard regular grade SD rat of selecting birth 1-3d, is provided by Lukang Medical Co., Ltd., Shandong's Experimental Animal Center, meets national medical animal and uses standard, and standard No. is clean level.
The former culture of 1.2SD rat retina neurocyte and the foundation of high sugared model; comprise the following steps that (concrete operations are referring to " levocarnitine is cultivated protective effect and the Mechanism Discussion of Rat retina neurocyte to external high sugar "; Chen Jie, University Of Qingdao's Master's thesis, 2013):
1.2.1 prepare following main agents
Containing 10% hyclone culture fluid, glucose, the bromo-2 ' BrdU of 5-, 4% paraformaldehyde, PBS phosphate buffer.
1.2.2 poly-D-lysine is processed Tissue Culture Flask or culture plate
For promoting the adherent of the retinal neuronal cell cultivated, before inoculation, process Tissue Culture Flask or culture plate with poly-D-lysine in advance.Method: in testing the previous day, 25mm
2culture bottle adds 100 μ g/ml poly-D-lysine 2.0ml, at the bottom of uniform fold bottle, in 37 DEG C of incubators, spends the night, and absorbs poly-D-lysine in culture bottle, and PBS washes three times, is put in 37 DEG C of incubator inner dryings for subsequent use.The cell of planning to implement identified by immunofluorescence is inoculated in 6 orifice plates, places in advance the coverslip of 27mm × 29mm in culture plate, and method poly-D-lysine advanced processing described above.
1.2.3 the preparation of primary retinal neuronal cell suspension
The SD rat of birth 1-3d is dipped to the 5min that carries out disinfection in 75% ethanol, the aseptic eyeball of getting in superclean bench, wash 3 times in ice bath PBS (containing penicillin 100U/ml penicillin and 100 μ g/ml streptomycins), under microscope, after corneoscleral junction, about 1mm place makes annular incision, remove anterior ocular segment tissue and vitreous body, blunt separation goes out layer of retina,neuroepithelial, ice bath PBS rinsing 2 times.The layer of retina,neuroepithelial of taking-up is cut into about 1mm by micro-eye scissors
2the piece of tissue of size, is collected in centrifuge tube.Add 0.125% trypsin of approximately 10 times of volumes, 37 DEG C of digestion 10-15min.DMEM/F12 culture fluid containing 10% hyclone stops digestion, after suction pipe is blown and beaten into single cell suspension gently repeatedly, 400 order stainless steel filtering nets filter, the centrifugal 5min of 1000r/min, collecting cell precipitation, containing the DMEM/F12 culture fluid re-suspended cell of 10% hyclone.Cell counting count board carries out cell counting, and adjusting cell density is 1 × 10
6individual/ml.
1.2.4 the inoculated and cultured of primary retinal neuronal cell
After cell suspension prepares, be inoculated in to be equipped with in advance and be coated with in 6 orifice plates of poly-D-lysine coverslip or culture bottle that poly-D-lysine was coated with, be placed in 37 DEG C, 5% CO
2incubator is cultivated.When cultivate add when 24h final concentration be the bromo-2 ' BrdU of 5-of 20 μ g/ml to suppress the growth of non-neurocyte, when 48h, change liquid, after this every 2-3d changes liquid once.
1.2.5 the foundation of high sugared model
In 24 orifice plates, when cell culture 3d, select 30mmol/L glucose to continue to cultivate 48h, observation of cell form cell suspension inoculation.
1.3 experiment grouping and methods
1. normal cell cultivation group (matched group); 2. high sugared damage group (HG); 3. levocarnitine protection group (LC, 100 μ mol/L); 4. L-arginine protection group (L-Arg, 600 μ mol/L); 5. the coordinating protection group (LC+L-Arg-1) of levocarnitine (50 μ mol/L) and L-arginine (300 μ mol/L); 6. the coordinating protection group (LC+L-Arg-2) of levocarnitine (50 μ mol/L) and L-arginine (50 μ mol/L); 7. department of the association protection group (LC+L-Arg-3) of levocarnitine (30 μ mol/L) and L-arginine (300 μ mol/L).
1. group get primary retinal neuronal cell and normally cultivate; the retinal neuronal cell that 2. group is got high sugared model is cultivated; 3.-and 7. group is got the retinal neuronal cell of high sugared model and is added corresponding protective agent to cultivate, and incubation time carries out trypan blue exclusion experimental calculation cell survival rate in the time of 24h and 48h.Concrete steps are: first digest centrifugal collecting cell, according to the resuspended liquid re-suspended cell of suitable cell for cell concentration, draw the resuspended cell of 100 μ l to centrifuge tube, add 100 μ l Trypan Blue liquid, mix gently dyeing 3min.Draw the cell having dyeed on a small quantity, with cell counting count board counting, each sample is at least counted 500 cells.
Cell survival rate=(total cellular score one blue cell number)/total cellular score × 100%.
1.4 statistical procedures
Experimental data represents with Y ± s, and Y represents average.Adopt SPSS17.0 statistical software to carry out data analysis.Statistical method adopts one factor analysis of variance, and between group, multiple comparisons adopts LSD-t inspection, has statistical significance taking P<0.05 as difference, and P<0.01 is that difference has remarkable statistical significance.
1.5 experimental result
This experimental result (in table 1) shows: the lower retinal neuronal cell apoptosis rate of high sugar effect significantly raise (P<0.01), levocarnitine protection group apoptosis rate is lower than the sugared damage group of height (P<0.05), and prompting levocarnitine can suppress the apoptosis due to high sugar damage; But the coordinating protection group apoptosis rate of the levocarnitine of three kinds of mol ratios and L-arginine is starkly lower than high sugared damage group (P<0.01), prompting levocarnitine profit L-arginine has been brought into play the effect of Synergistic.
The impact of the retina neural damage of table 1 levocarnitine associating L-arginine on diabetic retinopathy
Note: * represents and normal group compares P<0.05, and * * represents and relatively P<0.01 of normal group; # represents and high sugar group compares <0.05, and ## represents and relatively <0.01 of high sugar group.
Experiment 2
The former culture of 2.1SD rat retina M ü ller cell; comprise the following steps that (concrete operations are referring to " levocarnitine is cultivated protective effect and the mechanism of Rat retina M ü ller cell to external high sugar "; Zhang Ying, University Of Qingdao's Master's thesis, 2013):
2.1.1 poly-D-lysine is processed Tissue Culture Plate
2.1.2 the preparation of primary retina cell suspension
Take out raw after the SD rat of 1～3d, be placed in 75% ethanol soaking disinfection and anesthesia, after 5～8min by its execution, under aseptic condition, take out eyeball, be placed in PBS solution rinsing 3 times, remove ball week fascia tissue, clean eyeball, after corneoscleral junction, 1mm makes annular incision, takes out cornea, crystalline lens and vitreous body, by layer of retina,neuroepithelial blunt separation from eyecup, layer of retina,neuroepithelial is shredded with micro-eye scissors, collect in centrifuge tube, centrifugal, remove part supernatant.
In centrifuge tube, add isopyknic 0.125% trypsin, piping and druming gently, in 37 DEG C of water-baths, digest 8～15min, fully piping and druming cell, add containing the DMEM/F12 culture fluid of 10% hyclone and stop digestion, 400 order stainless steel filtering nets are filtered in centrifuge tube, the centrifugal 5min of 1000r/min, abandon supernatant, collecting cell precipitation.
In cell precipitation, add DMEM/F12 culture fluid (90%DMEM/F12 culture medium 15mM HEPES, 10% hyclone, 2.5mM L-glutaminate, penicillin 100U/ml, streptomycin 100 μ/ml), repeatedly blow and beat with re-suspended cell.
2.1.3 the inoculation of primary retina cell
The purification of 2.2SD rat retina M ü ller cell is cultivated
Culture fluid in sucking-off culture bottle, PBS washing 1 time, adds 0.125% trypsin to cover the bottle end) digestion, under phase contrast microscope, observe, see that cellular contraction becomes after circle, part cell suspension, absorbs trypsin.Add containing the DMEM/F12 culture fluid of 10% hyclone and stop digestion, repeatedly blow and beat re-suspended cell.Cell counting, same to primitive cell culture.Subpackage cell suspension, to culture bottle, is placed in 37 DEG C of cell culture incubators and cultivates.So repeat, after 2～3 times, can obtain the basically identical retina M ü ller cell of cellular morphology.
The foundation of the high sugared model of 2.3 retina M ü ller cells
In 24 orifice plates, when cell culture 3d, select 30mmol/L glucose to continue to cultivate 48h, observation of cell form cell suspension inoculation.
2.4 experiment grouping and methods
1. normal cell cultivation group (matched group); 2. high sugared damage group (HG); 3. levocarnitine protection group (LC, 100 μ mol/L); 4. L-arginine protection group (L-Arg, 600 μ mol/L); 5. the coordinating protection group (LC+L-Arg-1) of levocarnitine (50 μ mol/L) and L-arginine (300 μ mol/L); 6. the coordinating protection group (LC+L-Arg-2) of levocarnitine (50 μ mol/L) and L-arginine (50 μ mol/L); 7. the coordinating protection group (LC+L-Arg-3) of levocarnitine (30 μ mol/L) and L-arginine (300 μ mol/L).
1. group get primary retina M ü ller cell and normally cultivate; the retina M ü ller cell that 2. group is got high sugared model is cultivated; 3.-and 7. group is got the retina M ü ller cell of high sugared model and is added corresponding protective agent to cultivate, and incubation time carries out trypan blue exclusion experimental calculation cell survival rate in the time of 24h and 48h.Concrete steps are:
0.125% trypsinization M ü ller cell, collecting cell, the centrifugal 3min of 1000r/min, abandons supernatant, gets the resuspended liquid re-suspended cell of 500 μ l cell.Get the resuspended cell of 100 μ l to centrifuge tube, add 100 μ l Trypan Blue liquid (2X), mix, dyeing 3min.Draw the cell after 100 μ l dyeing, blood counting chamber counting, living cell counting and dead cell number respectively, calculates cell survival rate.
Cell survival rate=(total cellular score one blue cell number)/total cellular score × 100%.
2.5 statistical procedures
Experimental data represents with Y ± s, and Y represents average.Adopt SPSS17.0 statistical software to carry out data analysis.Statistical method adopts one factor analysis of variance, and between group, multiple comparisons adopts LSD-t inspection, has statistical significance taking P<0.05 as difference, and P<0.01 is that difference has remarkable statistical significance.
2.6 experimental result
This experimental result (in table 2) shows: the lower retina M ü ller apoptosis rate of high sugar effect significantly raise (P<0.01), levocarnitine protection group M ü ller apoptosis rate is lower than the sugared damage group of height (P<0.05), and prompting levocarnitine can suppress the M ü ller apoptosis due to high sugar damage; But the coordinating protection group M ü ller apoptosis rate of the levocarnitine of three kinds of mol ratios and L-arginine is starkly lower than high sugared damage group (P<0.01), prompting levocarnitine and L-arginine have been brought into play the effect of Synergistic.
The impact of the retina M ü ller cell injury of table 2 levocarnitine associating L-arginine on diabetic retinopathy
Note: * represents and normal group compares P<0.05, and * * represents and relatively P<0.01 of normal group; # represents and high sugar group compares <0.05, and ## represents and relatively <0.01 of high sugar group.
Experiment 3
The foundation of 3.1 diabetes rat models, concrete operations are referring to " experimentation of levocarnitine to the protective effect of diabetic retinal tissue in rat ganglionic cell ", Yu Changhong etc., " Chinese Pharmacological Bulletin ", the 29th volume o. 11th in 2013:
Under laboratory standard condition, rat high sugar feed adaptability is fed, and it is modeling success (fasting 12h before test) above that fasting glucose reaches 16.7mmol/L.
3.2 experiment grouping and methods
1. Normal group; 2. diabetic model group; 3. levocarnitine protection group (LC, 200mg/Kg); 4. L-arginine protection group (L-Arg, 1200mg/Kg); 5. the coordinating protection group (LC+L-Arg-1) of levocarnitine (100mg/Kg) and L-arginine (600mg/Kg); 6. the coordinating protection group (LC+L-Arg-2) of levocarnitine (100mg/Kg) and L-arginine (100mg/Kg); 7. the coordinating protection group (LC+L-Arg-3) of levocarnitine (60mg/Kg) and L-arginine (600mg/Kg).
1. group get normal rat normal saline gavage, 700mg/Kg, once a day; 2. group get diabetic model group rat normal saline gavage, 700mg/Kg, once a day; The 3. one 7. group get diabetic model group rat oral gavage and give corresponding protective agent, after 6 weeks, put to death rat, get retina dyeing.Process is referring to " experimentation of levocarnitine to the protective effect of diabetic retinal tissue in rat ganglionic cell ", Yu Changhong etc., " Chinese Pharmacological Bulletin ", the 29th volume o. 11th in 2013,1.3_3 joint.Each layer of optical microphotograph Microscopic observation retina, nucleus is the apoptotic cell that is of brown yellow granule dyeing.
Apoptotic index AI=(apoptotic cell quantity/total cellular score amount) × 100%.
3.3 statistical procedures
Experimental data represents with Y ± s, and Y represents average.Adopt SPSS17.0 statistical software to carry out data analysis.Statistical method adopts one factor analysis of variance, and between group, multiple comparisons adopts LSD-t inspection, has statistical significance taking P<0.05 as difference, and P<0.01 is that difference has remarkable statistical significance.
3.4 experimental result
This experimental result (in table 3) shows: the rat retinal ganglion cell apoptosis rate of diabetic model group significantly raise (P<0.01), levocarnitine protection group rat retinal ganglion cell apoptosis rate is lower than the sugared damage group of height (P<0.05), and prompting levocarnitine can suppress the rat retinal ganglion cell apoptosis due to diabetes; But the levocarnitine of three kinds of mol ratios and the arginic coordinating protection group of L mono-rat retinal ganglion cell apoptosis rate are starkly lower than diabetic model group (P<0.01), prompting levocarnitine and L. arginine have been brought into play the effect of Synergistic.
The impact of the retinal ganglion cells apoptosis of table 3 levocarnitine associating L. arginine on diabetes rat
| Group (n=6) | AI index (%) |
| Normal group | 2.4±0.7 |
| Diabetic model group | 15.5±2.8** |
| LC | 10.8±3.O*# |
| L-Arg | 14.7±1.4 |
| LC+L-Arg-1 | 4.8±1.2*## |
| LC+L-Arg-2 | 5.1±1.5*## |
| LC+L-Arg-3 | 5.2±1.0*## |
Note: * represents and Normal group compares P<0.05, and * * represents and relatively P<0.01 of Normal group; # represents and diabetic model group compares <0.05, and ## represents and relatively <0.01 of diabetic model group.
Claims (9)
1. a pharmaceutical composition, is characterized in that, contains levocarnitine and L-arginine in described compositions, is used for the treatment of diabetic renal papillary necrosis nerve injury.
2. pharmaceutical composition according to claim 1, is characterized in that, the mol ratio of described levocarnitine and described L-arginine is 1: 1～10.
3. pharmaceutical composition according to claim 1, is characterized in that, the dosage form of described compositions is tablet, capsule, powder, granule, solution, Emulsion or suspensoid.
4. the application of levocarnitine associating L-arginine in the medicine of preparation treatment of diabetic retinopathy change nerve injury.
5. application according to claim 4, wherein said diabetic renal papillary necrosis nerve injury is retina neural damage or M ü ller cell injury.
6. application according to claim 4, the mol ratio of wherein said levocarnitine and described L-arginine is 1: 1～10.
7. application according to claim 6, the mol ratio of wherein said levocarnitine and described L-arginine is 1: 3～8.
8. application according to claim 6, the mol ratio of wherein said levocarnitine and described L-arginine is 1: 6.
9. application according to claim 4, wherein said levocarnitine and described L-arginine carry out administration with tablet, capsule, powder, granule, solution, Emulsion, suspensoid.
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| EA039287B1 (en) * | 2016-11-11 | 2021-12-28 | Общество с ограниченной ответственностью медицинский центр "М.Т.К." | Method for the treatment of ischemic heart disease |
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| CN101534817A (en) * | 2006-10-31 | 2009-09-16 | 株式会社太平洋 | Use for treating obesity and diabetes |
| CN101939000A (en) * | 2007-12-17 | 2011-01-05 | 佛罗里达大学研究基金会有限公司 | Be used for the treatment of outgrowth material of pathologic ocular angiogenesis and method |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101534817A (en) * | 2006-10-31 | 2009-09-16 | 株式会社太平洋 | Use for treating obesity and diabetes |
| CN101939000A (en) * | 2007-12-17 | 2011-01-05 | 佛罗里达大学研究基金会有限公司 | Be used for the treatment of outgrowth material of pathologic ocular angiogenesis and method |
Non-Patent Citations (2)
| Title |
|---|
| A. CARMO: "L-Arginine transport in retinas from streptozotocin diabetic rats:correlation with the level of IL-1b and NO synthase activity", 《VISION RESEARCH》 * |
| 于常红等: "左卡尼汀对糖尿病大鼠视网膜神经节细胞保护作用的实验研究", 《中国药理学通报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017085642A1 (en) * | 2015-11-17 | 2017-05-26 | Товарыство З Обмэжэною Видповидальнистю Мэдычный Центр "Мтк" | Pharmaceutical composition |
| EA039287B1 (en) * | 2016-11-11 | 2021-12-28 | Общество с ограниченной ответственностью медицинский центр "М.Т.К." | Method for the treatment of ischemic heart disease |
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| Publication number | Publication date |
|---|---|
| CN103948581B (en) | 2018-05-08 |
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