CN101361754B - Use of rare-earth element salt or pharmaceutical composition thereof in preparing medicine for treating or preventing diabetes and adiposis - Google Patents
Use of rare-earth element salt or pharmaceutical composition thereof in preparing medicine for treating or preventing diabetes and adiposis Download PDFInfo
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Abstract
The invention relates to an application of rare earth element salts in biological field. The rare earth element salts or composite thereof can promote expression of PC1 and PC2 in islet beta cells and hypothalamus in human bodies, effectively promote secretion of mature insulin, lower level of blood sugar in animal bodies, thus achieving the purpose of curing diabetes; in addition, the rare earth element salts can also promote generation and secretion of alpha-MSH, thus achieving the purpose of curing obesity.
Description
Technical field
The present invention relates to rare-earth element salt or the purposes of its pharmaceutical composition in biological field, thereby refer in particular to that rare-earth element salt or its pharmaceutical composition promote in the pancreaticβ-cell in vivo and hypothalamus in the expression of PC1, PC2 promote beta cell secretion mature insulin and increase the secretion of hypophysis and hypothalamic alpha-MSH thereby to have the purposes for the treatment of diabetes and obesity.
Background technology
The ionic radius of rare earth element ion is similar to calcium ion, belongs to the hard acid metalloid together, and the geometric configuration of its Coordinative Chemistry behavior and coordination compound etc. are similar, can replace the position of calcium in many Biochemical processes, becomes the agonist of calcium.Replacement still keeps the character of calcium in some process afterwards, but also can produce and the more diverse biochemical properties of calcium, thereby the title of ' super calcium ' is arranged.
Rare earth element enter in the body will with the aminoacid wide application, found that the Rare Earth Amino-Acid coordination compound has bactericidal effect.After the eighties, there is the author to utilize the paramagnetic rare-earth ion as the probe research conformation of peptide in solution and the coordination mode of peptide and metal ion, these specific structural informations are for the structure-function relationship of the biomacromolecules such as analysing protein and enzyme, and the effect characteristics of understanding rare earth ion and other metal ion (such as Ca2+) and biomacromolecule all have important reference value.In addition, rare earth element can also and protein, the biomacromolecule combinations such as nucleic acid change its conformation and function.
Some rare earth compound has been developed as the medicine of curing the disease.Some rare earth compound has been developed as the medicine of curing the disease.The Dakin solution of a kind of commodity Ceriform by name in 1906 is just sold in the European market, and its main chemical compositions is potassium ceric sulfate.Sedemesis. has been used as antiemetic in the middle of last century and has been used for clinical.Nineteen eighty-two Britain Martindale pharmacopeia 28 editions is recorded cerous nitrate as the treatment burn medicine.Fox and Monafo etc. find 2.2% cerous nitrate (Ce (NO
3)
36H
2O) and the compound recipe suspension of 1% silver sulfadiazine (AgSD), burn there is good curative effect.
Rare earth compound has important medical value aspect anticoagulation.Be called clinically Trombodym-sulfo group .gamma.-pyridinecarboxylic acid neodymium is the high rare earth compound anticoagulant of a hypotoxicity and anticoagulant.
The sulfosalicylic acid chemical compound of neodymium and mishmetal has the analgesic cooling effect of highly significant, than good with the commonly used analgesic paroxysmal pain medicine aspirin of metering.
Experiment by rabbit and monkey shows, lanthanum chloride can prevent the aorta and the coronary atherosclerosis that are caused by meals.
In addition, the sequestration thing of gadolinium (Gd (DTPA)) is to use to get NMR contrast agent the most widely.
Tolbutamide Gd coordination compound (Gd-D860) is the rare earth compounding of the antidiabetic drug tolbutamide (D860) that synthesizes, find that Gd-D860 has stronger blood sugar reducing function, infer that its mechanism of action is to stimulate the pancreaticβ-cell uelralante to control blood sugar level.Nie Yuxiu etc. give low dosage SmCl to diabetes rat
3(0.05mg/kg body weight/day) can obviously improve the level of serum insulin, blood sugar lowering after 1.5 months.Further the effect of the results show rare earth is because adjusting islet cells secretory function causes.The diabetes that cause at the insulin dysmaturity can not fundamentally reach the purpose for the treatment of.So far do not find also that a good differentiation pancreatic stem cells of inducing promotes the ripe insulin secretion method that increases of insulin simultaneously.
Gadolinium is promoting to be used for the treatment of purposes in fat alpha-MSH and the ACTH secretion there are no research report.
Summary of the invention
Main purpose of the present invention provides rare-earth element salt or its pharmaceutical composition promotes the purposes that prohormone convertase 1 and prohormone convertase 2 are expressed in the medicine in the preparation treatment.
Another purpose of the present invention provides rare-earth element salt or its pharmaceutical composition promotes mature insulin and short melanocyte to stimulate the purposes in the hormone secretion medicine in the preparation treatment.
Another object of the present invention provides rare-earth element salt or the purposes of its pharmaceutical composition in preparation treatment or prevent diabetes and obesity drug.
The inventor has now found that after deliberation, rare-earth element salt can make pancreatic stem cells, beta cell in the pancreas, prohormone convertase 1 (prohormone convertase 1 in the beta cell tumor of In vitro culture, abbreviation PC1), prohormone convertase 2 (prohormone convertase 2, abbreviation PC2) expression raises, and these two kinds of enzymes can impel proinsulin to be cracked into insulin, and the secretion of mature insulin is increased.They are not only at external increase insulin and the C-peptide (micromolecule polypeptide that the C-peptide discharges when being degraded to mature insulin for the proinsulin human, the mensuration of C-peptide is the ripe more special assay method of people's pancreaticβ-cell, because it is not subjected to the interference of the hyclone in the cell culture medium.) secretion, and high fat fed the therapeutical effect that the diabetic mice of inducing generation has blood sugar lowering.In the pancreas of mice, observe increasing of PC1, PC2, and in the serum of mice, observe the reduction of insulin precurosor and the secretion of the mature insulin that increases.
In addition, the inventor finds after deliberation, rare-earth element salt can make the expression of PC1 in the hypothalamus, PC2 raise, these two kinds of enzymes can also impel precursor proopiomelanocortin (POMC) enzymolysis of Several Active Peptides to generate thyroliberin (ACTH) and short melanocyte stimulates hormone (alpha-MSH), and hypophysis and hypothalamic alpha-MSH secretion is increased.And alpha-MSH can reduce food ration by to control weight.Induce obesogenous mice that slimming therapeutical effect is arranged to high fat nursing.
Wherein, above-mentioned rare-earth element salt is acylate or the inorganic acid salt of rare earth element.
Above-mentioned rare earth element includes but not limited to: lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu), gadolinium (Gd), terbium (Tb), dysprosium (Dy), holmium (Ho), erbium (Er), thulium (Tm), ytterbium (Yb), lutecium (Lu), yttrium (Y), scandium (Sc), preferred rare earth element is lanthanum (La), gadolinium (Gd), lutecium (Lu), most preferably gadolinium (Gd).
Above-mentioned organic acid is citric acid or acetic acid, and described mineral acid is sulphuric acid, hydrochloric acid, hydrobromic acid, Fluohydric acid. or hydroiodic acid.
Further, the preferred chloride of rare-earth element salt, bromide, iodide, the more preferably chloride of rare earth element, most preferably Gadolinium trichloride.
The present invention also provides a kind of rare-earth element salt or the application of its pharmaceutical composition in preparation treatment or prevent diabetes medicine, comprises giving diabetics with rare-earth element salt.Diabetes herein refer to insufficient insulin or the diabetes that produce without insulin, such as I type and type ii diabetes.
In addition, the present invention also provides a kind of rare-earth element salt or the application of its compositions in preparation treatment or prevention of obesity disease drug, comprises giving the obesity patient with rare-earth element salt.The inventor gives mice that high fat feeds the obesity of inducing generation with rare-earth element salt or its compositions, the body weight gain of finding mice is controlled, athero in the body and the degree of interior fat all be improved significantly, from experimental data, gadolinium salt is than the better effects if of other rare-earth element salt.
Therefore, the present invention is by utilizing rare-earth element salt to promote the expression of PC1, PC2, thereby promotes pancreaticβ-cell secretion mature insulin, so that blood glucose descends in the animal body, and impel hypophysis and hypothalamic alpha-MSH secretion to increase, thereby reach the purpose for the treatment of diabetes and obesity.
The specific embodiment
Embodiment 1:
Gadolinium trichloride promotes the PC1 of the beta cell oncocyte of In vitro culture, the expression of PC2, promotes insulin precurosor to the conversion of mature insulin, increases the secretion of mature insulin.
Mice beta cell oncocyte is that NIT-1 is by (behind RPMI 1640 (U.S. Hyclone company)+10% hyclone (FBS) subculture, with 5 * 10
5The density kind of individual cells/well is implanted in 6 orifice plates, plants after 24 hours, and cell attachment is in good condition, culture fluid is changed to the PRMI1640 culture medium of serum-free, and hunger added respectively following culture medium after making the cultured cells synchronization in 16 hours:
①RPMI 1640+10％FBS
②RPMI 1640+10％FBS+0.1mM GdCl3
③RPMI 1640+10％FBS+0.2mM GdCl
3
④RPMI 1640+10％FBS+0.5mM GdCl
3
⑤RPMI 1640+10％FBS+1.0mM GdCl
3
6. RPMI 1640+10% hyclone+1.0mM NaCl
Respectively at 37 ℃, CO
2Cultivate in the incubator of concentration 5% after 24 hours, extract whole-cell protein and do Western Blotting experimental analysis, detect the variation of PC1, PC2 expressing quantity.
Above six kinds of albumen samples, setting sample PC1, PC2 expressing quantity 1. is with reference to " 1 ".In the situation that Tot Prot identical (β-actin expression is consistent) is observed in the situation that normal saline does not change PC1, PC2 expressing quantity, along with the GdCl that acts on cell
3The increase of concentration, PC1, PC2 expressing quantity also increase gradually.The results are shown in Table 1:
Table 1.GdCl
3Inducing mouse beta cell oncocyte is the PC1 of NIT-1, the variation of PC2 expressing quantity
| ① | ② | ③ | ④ | ⑤ | ⑥ | |
| PC1 | 1.0 | 2.3 | 3.2 | 6.5 | 8.8 | 1.0 |
| PC2 | 1.0 | 2.7 | 2.9 | 3.8 | 4.9 | 1.0 |
Extract simultaneously each sample RNA, in the situation that concentration identical (18s RNA is as confidential reference items), through the mRNA content of Real Time PCR detection PC1, PC2, the result shows: along with the GdCl that acts on cell
3The increase of concentration, the content of the mRNA of PC1, PC2 also increases gradually, shows GdCl
3Stimulated the expression of PC1, PC2 on the transcriptional level.
Table 2.GdCl
3Inducing mouse beta cell oncocyte is the PC1 of NIT-1, the variation of PC2mRNA
| ① | ② | ③ | ④ | ⑤ | ⑥ | |
| PC1 | 1.0 | 9.1 | 24.5 | 60.9 | 97.4 | 1.0 |
| PC2 | 1.0 | 5.6 | 16.2 | 31.3 | 80.0 | 1.0 |
Above condition of culture, suck culture fluid, measure the interior insulin of born of the same parents and the content of C-peptide, or this cell is carried out sugared stress tests, namely suck culture fluid, add the RPMI RPMI-1640 that contains in right amount glucose 2.5mM, after 90 minutes, culture fluid is changed into the RPMI RPMI-1640 that contains glucose 25mM, after 90 minutes, collect respectively culture fluid and cell, measure the difference content of its insulin and C-peptide.The result represents: after rare earth element is induced, the insulin in the mice B-glucagonoma cell line NIT-1 cell born of the same parents and the content of C-peptide with all increase to some extent, and sugar stress after, the insulin in the born of the same parents and the release of C-peptide all be improved (table 3, table 4).
Mice beta cell oncocyte was the interior insulin of NIT-1 cell born of the same parents and the content of C-peptide before and after table 3. was induced
| Insulin (μ IU/ml) | C-peptide (pmol/ml) | |
| Before inducing | 20.7 | 15.7 |
| After inducing (0.01mM Gadolinium trichloride) | 43.8 | 24.3 |
| After inducing (0.1mM Gadolinium trichloride) | 86.0 | 38.6 |
| After inducing (1mM Gadolinium trichloride) | 111.2 | 45.4 |
Mice beta cell oncocyte was NIT-1 cell sugar stress tests result before and after table 4. was induced
| Insulin content in the cell culture fluid (μ IU/ml) | Insulin content (μ IU/ml) in the cell born of the same parents | |
| Before inducing | 21.5 | 20.7 |
| After inducing (0.01mM Gadolinium trichloride) | 43.3 | 43.8 |
| After inducing (0.1mM Gadolinium trichloride) | 85.7 | 86.0 |
| After inducing (1mM Gadolinium trichloride) | 119.5 | 111.2 |
Embodiment 2:
Separation and purification normal mouse islets of langerhans, external through GdCl
3The variation of PC1, PC2 expression after stimulating
Only choose 6-8 week macrandry ICR/C57 mice 6-10, the separating-purifying islets of langerhans, with the islets of langerhans In vitro culture that extracts, the separation and purification concrete grammar of mouse islets is as follows:
1. mice overnight fasting (can't help water), the execution of craning one carries out disinfection to skin of abdomen with 75% ethanol.
2. carefully open the abdominal cavity, blunt separation fully exposes gallbladder.Enter duodenal position at common bile duct and carry out ligation with suture.
3. syringe is carefully inserted 2～3ml/ mice of By Injection of Collagenase working solution from the nearly liver end of common bile duct.Until pancreas is fully full expand after, win fast pancreas, insert in the clean centrifuge tube, add the 3ml collagenase.
4. centrifuge tube is put into 37 ℃ of water-baths and digested, frequently jolt, pancreas is fully contacted with collagenase.
5. when treating pancreas digestion for fine sand shape (approximately needing about 30 minutes), take out, add immediately cold HBSS or PBS fully to jolt mixing, leave standstill to stop digestion on ice in 4 ℃.
6. after Digestive system cools off fully, in 4 ℃, centrifugal 30 seconds of 1000rpm.Abandon supernatant, add cold HBSS or the PBS of 10ml and fully jolt mixing, in 4 ℃, centrifugal 30 seconds of 1000rpm.
7. with the supernatant Ex-all, add 5ml Histopaque1077, fully mixing is suspension, carefully adds 5mlHBSS or PBS again, is sure not to jolt test tube.4 ℃, centrifugal 20 minutes of 2200rpm is necessary for brakeless and stops.At this moment, liquid is divided into three layers in the visible pipe, with second layer sucking-off, places another clean centrifuge tube, repeatedly washes 2-3 time with HBSS or PBS.
8. in microscopically islets of langerhans is chosen, in (1640+15%FBS), cultivated.Islets of langerhans can be dyeed specifically by dithizone (DTZ), is scarlet.The preparation of collagenase working solution: Collagenase V (available from Sigma) is dissolved in the working solution that is made into concentration 1mg/ml among HBSS or the PBS, and filtration sterilization is stored in 4 ℃ of refrigerators for subsequent use.Matching while using is placed and is no more than 2 days as far as possible.
Separation and purification normal mouse islets of langerhans is divided into 3 groups at random, places respectively
①RPMI 1640+15％FBS
②RPMI 1640+15％FBS+1.0mM GdCl
3
③RPMI 1640+15％FBS+1.0mM NaCl
In three kinds of culture medium, in 37 ℃, cultivate after 24 hours in the incubator of CO2 concentration 5%, collect three groups whole protein lysate, detect the difference of three groups of sample P C1, PC2 expressing quantity with Western Blotting method.Found that, in the situation that Tot Prot identical (GAPDH is as confidential reference items albumen), through GdCl
3Post-stimulatory islets of langerhans PC1, PC2 expressing quantity are all than the high 4-6 of islets of langerhans that does not stimulate doubly.And the islets of langerhans that gives NaCl does not obviously change with the normal islets of langerhans of cultivating.
Extract simultaneously three groups mRNA, detect the mRNA content difference of three groups of sample nucleic acid level PC1, PC2 through the RealTime PCR method.Found that, in the situation that mRNA equivalent (18s RNA is as confidential reference items), through GdCl
3Islets of langerhans PC1, the PC2 that stimulates be do not stimulate islets of langerhans 4-6 doubly.And the islets of langerhans that gives NaCl does not obviously change with the normal islets of langerhans of cultivating.The results are shown in Table 5,6.
The variation of the level of PC1, the PC2mRNA of mouse islets before and after table 5. is induced
| Before inducing | After inducing | The normal saline contrast | |
| PC1 | 1.0 | 4.8 | 1.0 |
| PC2 | 1.0 | 5.1 | 1.0 |
The variation of the level of the PC1 of mouse islets, PC2 albumen before and after table 6. is induced
| Before inducing | After inducing | The normal saline contrast | |
| PC1 | 1.0 | 3.3 | 1.0 |
| PC2 | 1.0 | 4.6 | 1.0 |
Embodiment 3:
GdCl
3Therapeutic effect to the diabetic mice of Pax6 gene mutation
There is serious abnormal development in Pax6 homozygous mutation mice, can't survive, so we selects each several of the heterozygous mutant mouse male and female of child-bearing period, copulation production.Newborn mice is given birth to normal ablactation in rear 21 days, and the male mice (comprising that genotype is normal and heterozygous mutant) of choosing wherein gives the high lipid food nursing.
Monitoring mouse blood sugar from high fat fed for 6 weeks found that Pax6 mutant mouse majority feeds 10 all future trouble diabetes at high fat.The results are shown in Table 7:
Table 7.Pax6 homozygous mutation diabetes mice model mice
Annotate: 1. respectively organize the mice number all greater than 20.
2. high lipid food is provided by Department Of Medicine, Peking University's Animal House, prescription: 21% fat, and 0.15% cholesterol, all the other compositions and normal diet are close.
3. lumbar injection carbohydrate tolerance test IPGTT (Intraperitoneal Glucose Tolerance Testing) is as the index that detects mice trouble diabetes.Method is as follows:
1. IPGTT detects 5-6 point survey evening before yesterday Mouse Weight, and fasting, can't help water, change dressings, the impact of removing residual foodstuff;
2. after fasting 15-16 hour, namely morning next day the 8-9 point, row IPGTT test.Cut 5-6 μ m from Mouse Tail-tip and get tail blood survey fasting glucose, after the aseptic cotton carrier hemostasis, static 15 minutes;
3. mouse peritoneal is injected 20% glucose, and volume calculates by 10 μ l/g body weight, reduces the stimulation to mice in the operation as far as possible;
4. surveyed 30 minutes after the injection, 60 minutes, 120 minutes blood glucose;
5. add the pan feeding piece after experiment is finished.
Glucose for injection: anhydrous glucose is dissolved in sterilized water and is made into 20% glucose, and filtration sterilization is stored in 4 ℃ of refrigerators for subsequent use.Rapid blood sugar instrument and reagent paper are Johnson ﹠ Johnson's product.
Choose Pax6 sudden change diabetic mice, observe GdCl
3Hypoglycemic effect.
Diabetic mice is divided into two groups: first group is GdCl
3Treatment group, second group is physiology saline control group.Treatment is carried out synchronously by two groups, and administering mode is lumbar injection, is administered once in per four days, and the per injection volume calculates by 10 μ l/g body weight, the change of blood sugar of per 16 days capable IPGTT monitoring mices.Treatment GdCl
3Preparation: with solid GdCl
3Powder (available from Sigma) is dissolved in aquesterilisa, is made into the GdCl that concentration is 10mM
3Solution, filtration sterilization is stored in 4 ℃ of refrigerators for subsequent use.Normal saline is injection 0.9% sodium chloride solution, sterilizes, and is stored in 4 ℃ of refrigerators for subsequent use.
Found that: after experiment was carried out 32 days, the treatment group mouse blood sugar all had reduction, wherein 10% has returned to the euglycemia level, and 70% mice is IGT (impaired glucose tolerance), and control group mice blood glucose obviously raises, aggravation.After experiment is carried out 64 days, take IPGTT120 minute blood glucose as standard, estimate the carbohydrate metabolism level of two groups of mices, the treatment group mouse blood sugar reduces 30%-40%, simultaneously control group mice blood sugar increasing 40%-50%.And in the discovery experimentation, the body weight gain for the treatment of group mice is controlled, and the body weight of control group mice rises always.Find in the process of drawing materials, the athero in the treatment group Mice Body and the degree of interior fat are compared with matched group, all be improved significantly.Concrete outcome sees Table 8:
The blood sugar level of experiment diabetic mice before and after table 8. treatment
| Blood sugar concentration (mM) | |
| Before the treatment | 14.9±4.3 |
| GdCl 3Treatment group | 9.7±3.6 |
| The normal saline matched group | 19.5±5.4 |
Annotate:
Below respectively organize the mice number all more than 10.
The present WHO of diabetes diagnosis standard application, the ADA universal standard.
Embodiment 4:
The diabetic mice GdCl of Pax6 sudden change
3The variation of PC1, PC2 after the treatment
Two groups of mouse islets among the embodiment 3 are carried out separating-purifying, collect the whole protein lysate of two groups of islets of langerhans, detect the difference of two groups of sample P C1, PC2 expressing quantity with the WesternBlotting method.
Found that, in the situation that Tot Prot identical (GAPDH is as confidential reference items albumen), treatment group islets of langerhans PC1, PC2 expressing quantity are than the high 4-6 of matched group times, and albumen is more stable.Extract the RNA of two groups of islets of langerhans, after reverse transcription is cDNA, detect the difference of the mRNA level of two groups of sample P C1, PC2 through the RealTime PCR method.In the situation that RNA equivalent (18s RNA is as confidential reference items), the mRNA amount for the treatment of group islets of langerhans PC1 is 5-7 times of matched group, and the amount for the treatment of group islets of langerhans PC2 is 70-100 times of matched group.
The comparison of islets of langerhans form: the mouse islets of separating-purifying, in microscopically counting and size relatively, find that the islets of langerhans number for the treatment of group mice is about more than 5 times of matched group, and volume is obviously greater than matched group, and complete form.
Embodiment 5:
GdCl
3The diabetic mice serum insulin precursor of Pax6 sudden change and the variation of mature insulin after the treatment
The manufacture method of diabetic mice and GdCl
3Therapeutic Method with embodiment 3.
The result shows, sugar stimulation GdCl after 2 hours
3The content of mature insulin obviously raises in the treatment group serum, and the control group mice insulin precurosor of normal saline treatment is higher than treatment group, shows GdCl
3Treatment has promoted maturation and the secretion of the insulin in the diabetic mice body, thereby reaches the effect of blood sugar lowering treatment diabetes. concrete outcome sees Table 9.
The serum insulin level of table 9. experiment diabetic mice
| Insulin precurosor content (pM) | Insulin content (pM) | |
| Normal group | 22.0±0.12 | 151.8±6.39 |
| Diabetic model group | 73.6±2.63 | 130.12±1.00 |
| Diabetes+GdCl 3Treatment group | 20.3±0.58 | 158.4±8.32 |
Embodiment 6:
The citric acid gadolinium promotes the PC1 of the B-glucagonoma cell of In vitro culture, the expression of PC2, promotes insulin precurosor to the conversion of mature insulin, increases the secretion of mature insulin.
Experimental technique is with embodiment 1.The result is as follows:
Table 10. citric acid gadolinium inducing mouse beta cell oncocyte is the PC1 of NIT-1, the variation of PC2 expressing quantity
| ① | ② | ③ | ④ | ⑤ | ⑥ | |
| PC1 | 1.0 | 2.5 | 3.6 | 7.2 | 7.9 | 1.0 |
| PC2 | 1.0 | 1.9 | 2.6 | 5.8 | 6.3 | 1.0 |
Table 11. citric acid gadolinium inducing mouse beta cell oncocyte is the PC1 of NIT-1, the variation of PC2mRNA
| ① | ② | ③ | ④ | ⑤ | ⑥ | |
| PC1 | 1.0 | 8.2 | 16.5 | 33.9 | 68.2 | 1.0 |
| PC2 | 1.0 | 4.7 | 14.2 | 29.6 | 57.0 | 1.0 |
Mice beta cell oncocyte was the interior insulin of NIT-1 cell born of the same parents and the content of C-peptide before and after table 12. citric acid gadolinium was induced
| Insulin (μ IU/ml) | C-peptide (pmol/ml) | |
| Before inducing | 21.3 | 14.9 |
| After inducing (0.01mM citric acid gadolinium) | 39.9 | 22.8 |
| After inducing (0.1mM citric acid gadolinium) | 79.1 | 33.5 |
| After inducing (1mM citric acid gadolinium) | 105.8 | 42.3 |
Embodiment 7:
Gadolinium is to the treatment of diabetic mice
Experimental technique is with embodiment 3.The result is as follows:
Blood glucose and the serum insulin level of table 13. experiment diabetic mice
| Blood glucose (mM) | Insulin content (μ IU/ml) | |
| Diabetic model group | 15.6±2.74 | 9.12±1.00 |
| Diabetes+gadolinium group | 8.9±1.96 | 16.9±3.64 |
Embodiment 8:
Digadolinium trisulfate is to the treatment of diabetic mice
Experimental technique is with embodiment 3.The result is as follows:
Blood glucose and the serum insulin level of table 14. experiment diabetic mice
| Blood glucose (mM) | Insulin content (μ IU/ml) | |
| Diabetic model group | 15.6±2.74 | 9.12±1.00 |
| Diabetes+Digadolinium trisulfate group | 11.3±3.51 | 14.2±2.23 |
Embodiment 9:
Lanthanum chloride is to the treatment of diabetic mice
Experimental technique is with embodiment 3.The result is as follows:
Blood glucose and the serum insulin level of table 15. experiment diabetic mice
| Blood glucose (mM) | Insulin content (μ IU/ml) | |
| Diabetic model group | 15.6±2.74 | 9.12±1.00 |
| Diabetes+lanthanum chloride group | 12.2±2.12 | 11.9±3.05 |
Embodiment 10:
Lutecium chloride is to the treatment of diabetic mice
Experimental technique is with embodiment 3.The result is as follows:
Blood glucose and the serum insulin level of table 16. experiment diabetic mice
| Blood glucose (mM) | Insulin content (μ IU/ml) | |
| Diabetic model group | 15.6±2.74 | 9.12±1.00 |
| Diabetes+lutecium chloride group | 13.7±1.28 | 11.1±2.66 |
Embodiment 11:
GdCl
3The variation of PC1, PC2 in the obesity mice hypothalamus that the high fat of Pax6 sudden change is fed after the treatment
Two groups of mice thalamuses among the embodiment 3 are carried out separating-purifying, collect the whole protein lysate of two groups of thalamuses, detect the difference of two groups of sample P C1, PC2 expressing quantity with the WesternBlotting method.
Found that, in the situation that Tot Prot identical (GAPDH is as confidential reference items albumen), treatment group thalamus PC1, PC2 expressing quantity are than the high 2-3 of matched group times, and albumen is more stable.Extract the RNA of two groups of thalamuses, after reverse transcription is cDNA, detect the difference of the mRNA level of two groups of sample P C1, PC2 through the RealTime PCR method.In the situation that RNA equivalent (18sRNA is as confidential reference items), the mRNA amount for the treatment of group thalamus PC1 is 10-12 times of matched group, and the amount for the treatment of group islets of langerhans PC2 is 60-80 times of matched group.
Embodiment 12:
GdCl
3The variation of alpha-MSH and ACTH content in the serum of the obesity mice that the high fat of Pax6 sudden change is fed after the treatment
Collect two groups of mice serums among the embodiment 3, detect two groups of sample alpha-MSH and ACTH content with test kit.
Found that, alpha-MSH content is higher 3.5 times than matched group in the treatment group mice serum, and ACTH content is higher 3.3 times than matched group.
Embodiment 13:
GdCl
3The body weight change of the obesity mice that the high fat of Pax6 sudden change is fed after the treatment
The high fat of table 17. is fed the contrast (the treatment precursor is reset to 1) of the treatment front and back body weight of mice
| GdCl 3Treatment group | The normal saline matched group | |
| Before the treatment | 1 | 1 |
| GdCl 3Treated rear 16 days | 0.994 | 1.098 |
| GdCl 3Treated rear 32 days | 1.030 | 1.137 |
| GdCl 3Treated rear 48 days | 1.089 | 1.228 |
| GdCl 3Treated rear 64 days | 1.129 | 1.329 |
Embodiment 14:
The citric acid gadolinium, gadolinium, Digadolinium trisulfate, lanthanum chloride, the body weight change of the obesity mice that the high fat of Pax6 sudden change is fed after the lutecium chloride treatment
The high fat of table 18. is fed the contrast (the treatment precursor is reset to 1) of the treatment front and back body weight of mice, and treatment time is 64 days
| The normal saline matched group | Citric acid gadolinium treatment group | The Digadolinium trisulfate treatment group | The lanthanum chloride treatment group | The lutecium chloride treatment group | |
| Before the treatment | 1 | 1 | 1 | 1 | 1 |
| After the treatment | 1.329 | 1.114 | 1.143 | 1.198 | 1.207 |
In sum, rare-earth element salt can effectively promote the secretion of mature insulin, so that blood glucose descends in the animal body, reaches the effect for the treatment of diabetes; And can promote generation and the secretion of alpha-MSH, thereby reach the purpose for the treatment of of obesity.Wherein from experimental data, gadolinium salt is more excellent than other rare earth element effect.
Claims (6)
1. rare-earth element salt or its Pharmaceutical composition purposes in preparation treatment or prevention of obesity disease drug, it is characterized in that, described rare-earth element salt is acylate or the inorganic acid salt of rare earth element, and described rare earth element is lanthanum (La), gadolinium (Gd), lutecium (Lu).
2. purposes according to claim 1 is characterized in that, described rare earth element is gadolinium (Gd).
3. purposes according to claim 1 is characterized in that, described organic acid is citric acid or acetic acid, and described mineral acid is sulphuric acid, hydrochloric acid, hydrobromic acid, Fluohydric acid. or hydroiodic acid.
4. purposes according to claim 1 is characterized in that, described rare-earth element salt is the chloride of rare earth element, bromide, iodide.
5. purposes according to claim 4 is characterized in that, described rare-earth element salt is the chloride of rare earth element.
6. purposes according to claim 5 is characterized in that, described rare-earth element salt is Gadolinium trichloride.
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| CN110279683A (en) * | 2019-07-04 | 2019-09-27 | 江苏安唐医药科技有限公司 | Pharmaceutical composition and its application in preparation treatment and prevention diabetes medicament |
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