CN114948989A - Application of magnesium hydride in preparing medicine for preventing and treating kidney crystal lithiasis - Google Patents
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Abstract
The invention provides an application of magnesium hydride in preparing a medicament for preventing and treating renal calculus, wherein the medicament takes magnesium hydride as a unique active ingredient, or a medicinal composition containing magnesium hydride. The invention discloses the application of magnesium hydride in preparing the medicine for preventing and treating the kidney crystal lithiasis for the first time, the magnesium hydride has great application potential in crystallizing the kidney injury as a brand-new treatment means, and has important significance and wide clinical application prospect in the research of curative effect evaluation.
Description
Technical Field
The invention relates to the technical field of medicines for preventing and treating renal calculus, in particular to application of magnesium hydride in preparing medicines for preventing and treating renal calculus and a related pharmaceutical composition.
Background
The kidney stones are common diseases of the urinary system, are widely distributed in the global range, bring heavy economic and medical burden to the country, particularly Asia, the prevalence rate of China is as high as more than 4.87-10%, and the prevalence rate of China is on the trend of increasing year by year, along with the development of surgical technology, the treatment means of the kidney stones are gradually enriched (such as external shock wave lithotripsy, percutaneous nephrolithotomy and the like), but the recurrence rate is still high, statistics shows that the five-year recurrence rate exceeds 50%, and at present, no effective method for preventing generation and recurrence of the stones is available.
Hydrogen, which is the simplest and most abundant element in the natural world, has been widely noticed and studied in various fields since japanese scientists have found that hydrogen can treat diseases by selective antioxidation, and has been found to exhibit preventive and therapeutic effects in various disease models and human diseases, and that hydrogen exerts its selective antioxidation action by direct or indirect action, and scavenging harmful free radicals in the body to alleviate body damage is currently considered to be the most important form in which hydrogen exerts its action. However, the hydrogen intake method has certain limitations, such as the intake of hydrogen or the intake of hydrogen-rich water, and thus the biological effect of the solid hydrogen carrier becomes a hot spot in hydrogen medicine. Magnesium Hydride (MgH) 2 ) Is a novel high-efficiency magnesium-based hydrogen storage material, has large hydrogen release amount and byproducts Mg (OH) 2 No harm to human body, etc. The characteristics that 1 g of magnesium hydride can generate 1729 ml of hydrogen and hydrolysis slowly releases the hydrogen for a long time make the magnesium hydride an ideal biological hydrogen supply material. Magnesium hydride was first used in biological studies by Kamimura N in 2016, and the results suggest that magnesium hydride can improve plasma triglyceride levels and prolong the life expectancy of wild-type mice fed a high fat diet. The latest research on magnesium hydride comes from korea, and the results suggest that magnesium hydride prevents the occurrence of vascular dementia by protecting hippocampal neurons.
Disclosure of Invention
In order to overcome the technical defects, the first aspect of the invention provides application of magnesium hydride in preparing a medicament for preventing and treating renal calculus.
The renal crystal lithiasis provided by the invention specifically refers to renal crystal lithiasis and renal lithiasis;
the prevention and treatment of the invention refers to prevention or treatment, wherein the prevention refers to prevention of renal crystallization transformation into renal stone disease.
Furthermore, the medicine takes magnesium hydride as the only active component, or contains a magnesium hydride medicinal composition.
Furthermore, the content of magnesium hydride in the medicine is 0.1-99 wt%. Preferably 0.5 to 10 wt%.
Further, the pharmaceutical composition also comprises pharmaceutically acceptable auxiliary materials.
Further, the pharmaceutically acceptable auxiliary materials are selected from one or more than two of corn oil, glycerol, propylene glycol and water.
Further, the medicine is an oral preparation or an injection preparation.
The second aspect of the invention provides an oral preparation for preventing and treating kidney crystal lithiasis, which comprises magnesium hydride and pharmaceutically acceptable auxiliary materials. Preferably, the pharmaceutically acceptable auxiliary material is a solvent, preferably, the solvent is oil, and preferably, the oil is corn oil. Preferably, the solvent can also be glycerol, propylene glycol, water or the like.
The pharmaceutically acceptable auxiliary materials refer to conventional pharmaceutical auxiliary materials in the pharmaceutical field, wherein diluents and excipients are water and the like; binders such as cellulose derivatives, gelatin, or polyvinylpyrrolidone, etc.; fillers such as starch and the like; disintegrating agents such as calcium carbonate or sodium bicarbonate; other adjuvants such as flavoring agents and/or sweetening agents may also be added to the composition.
The pharmaceutical composition can be prepared into various dosage forms by adopting a conventional method in the medical field and taking magnesium hydride as an active ingredient and pharmaceutically acceptable auxiliary materials. Can be applied to individuals needing treatment by oral administration, intraperitoneal injection, subcutaneous injection, intravenous injection, intramuscular injection, lymph node injection, mucosal administration and other ways according to dosage forms. The individual may be a human or an animal. The dosage is generally 1-1000 mg/kg body weight/day, and can be changed according to age, disease condition and the like of an individual. When used for oral administration, it can be prepared into conventional solid preparations such as tablets, powders, capsules, etc.; for injection, it can be prepared into injection. In various formulations, the magnesium hydride active ingredient is present in an amount of 0.1% to 99% by weight, preferably 0.5 to 10% by weight.
After the technical scheme is adopted, compared with the prior art, the method has the following beneficial effects:
the invention discloses the application of magnesium hydride in preparing the medicine for preventing and treating the kidney crystal lithiasis for the first time, the magnesium hydride has great application potential in the crystal renal injury as a brand-new treatment means, and has important significance and wide clinical application prospect in the research of curative effect evaluation.
Drawings
FIG. 1 is a graph of various concentrations of magnesium hydride corn oil formulations. The magnesium hydride corn oil preparation with the concentration of 12.5mg/ml, 25mg/ml, 50mg/ml and 100mg/ml is subjected to macroscopic property expression after standing for 60 minutes;
FIG. 2 is a graph of the dose effect of magnesium hydride to ameliorate glyoxylate induced kidney tissue damage in a kidney mouse model. Con group is prepared by injecting normal saline into abdominal cavity for 5 days, Gly group is prepared by injecting glyoxylate into abdominal cavity for 5 days, and Gly + MH50 group, Gly + MH100 group, Gly + MH200 group, and Gly + MH400 group are prepared by respectively administering MgH 2 days in advance 2 50Mg/kg, 100Mg/kg, 200Mg/kg and 400Mg/kg, and continuously performing intraperitoneal injection on glyoxylate for 5 days after 2 days, wherein the interval between the two medicines is 4-6 hours, and the magnification is as follows: 200 times of the total weight of the powder;
FIG. 3 is a graph of magnesium hydride improving the degree of renal tubular injury. Con group is prepared by intraperitoneal injection of normal saline with equal dosage for 5 days, Gly group is prepared by intraperitoneal injection of glyoxylate for 5 days, and GH group is prepared by administering MgH 2 days earlier 2 Gavage 200mg/kg, performing intraperitoneal injection of glyoxylate for 5 days, separating two medicines by 4-6 hours, and performing injection on GOH group 2 days earlier with Mg (OH) 2 Performing intragastric administration, and continuing to perform intraperitoneal injection on the glyoxylate for 5 days after 2 days, wherein the interval between the two medicines is 4-6 hours; GV group is administered 2 days earlier to cornThe oil is infused into the stomach, and the glyoxylate is continuously injected into the abdominal cavity for 5 days after 2 days, and the interval time between the two is 4 to 6 hours. FIG. 3 shows kidney tissue H of five mice&E. Von kossa, Masson staining, magnification: 200 times of the total weight of the powder;
figure 4 is the renal function of mice with magnesium hydride-protected crystalline kidney injury, # p <0.05 (relative to Con group), # p <0.05 (relative to Gly group);
figure 5 is a graph showing that magnesium hydride reduces the level of inflammation in crystallized kidney injured mice, # p <0.05 (relative to Con group), # p <0.05 (relative to Gly group);
FIG. 6 shows that magnesium hydride improves renal tissue oxidative stress and lipid peroxidation levels in mice with renal injury, # p <0.05 (relative to Con group) and # p <0.05 (relative to Gly group);
FIG. 7 shows that magnesium hydride improves tubular cell apoptosis in crystallized kidney-injured mice. Grouping and dosing were as in figure 3, and figure 7 is TUNEL staining of five groups of mice with renal tissue responsive to apoptosis, stained with apoptotic cells as indicated by the red arrows, at 400-fold magnification.
Detailed Description
The advantages of the invention are further illustrated in the following description of specific embodiments in conjunction with the accompanying drawings. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
Preparation and application of magnesium hydride corn oil preparation
The test method comprises the following steps: magnesium hydride (purchased from Wuhankemike biomedical technologies, Inc.) is added into corn oil for experiment (glycerol, propylene glycol, water and other reagents can be used as the pharmaceutical carrier of magnesium hydride according to the experiment requirements), and magnesium hydride corn oil preparations with the concentrations of 12.5mg/ml, 25mg/ml, 50mg/ml and 100mg/ml are prepared according to the experiment requirements. After the preparation is sufficiently vortexed and shaken, the magnesium hydride is further uniformly distributed in the corn oil by using ultrasound. The magnesium hydride corn oil preparation is required to be ready for use, and if an aqueous reagent is used as a magnesium hydride carrier, it is required to complete administration within a short time immediately after preparation.
And (3) test results: the prepared magnesium hydride corn oil preparation can keep stable physicochemical properties for a relatively long time under normal temperature and normal pressure, and can ensure the uniformity and stability of animal administration, the magnesium hydride corn oil preparation with different concentrations is shown in figure 1, and the magnesium hydride corn oil preparation with different concentrations before standing or after standing for 60 minutes is shown as a suspension with uniform state in figure 1; wherein, the low-concentration (when 12.5mg/ml, 25mg/ml and 50 mg/ml) magnesium hydrate corn oil preparation before standing is a light yellow uniform suspension, the light yellow gradually becomes lighter along with the increase of the concentration, and when the concentration is increased to 100mg/ml, the magnesium hydride corn oil preparation is a milky white suspension; the color and suspension state of the magnesium hydride corn oil preparation after standing for 60 minutes hardly change, which indicates that the physical and chemical properties of the magnesium hydride corn oil preparation are stable. Meanwhile, the hydrolysis reaction of the magnesium hydride protected by the corn oil can be slowed down after the magnesium hydride enters the body, and the magnesium hydride is prevented from rapidly reacting in a gastric acid strong acid environment, so that the time of hydrogen release in the magnesium hydride body is prolonged.
Dose-effect relationship of magnesium hydride for treating crystalline renal injury
The test method comprises the following steps: a crystallized kidney injury model of a C57BL/6 mouse is constructed by using a glyoxylate intraperitoneal injection (100mg/kg, 5 days of intraperitoneal injection), a Con group is a normal saline with equal dosage for intraperitoneal injection for 5 days, a Gly group is used for intraperitoneal injection for 5 days, and a Gly + MH50 group, a Gly + MH100 group, a Gly + MH200 group and a Gly + MH400 group are respectively given to MgH 2 days in advance 2 50Mg/kg, 100Mg/kg, 200Mg/kg and 400 Mg/kg. Respectively using 50, 100, 200 and 400mg/kg MgH 2 days in advance 2 Performing intragastric administration, continuing to perform the glyoxylate intraperitoneal injection for 5 days after 2 days, and performing the glyoxylate intraperitoneal injection twice at an interval of 4-6 hours. After mice receive glyoxylate and are injected into abdominal cavities for 5 days, kidney tissues of each group of mice are taken and fixed by paraformaldehyde, and after the steps of dehydration, embedding, slicing and the like, the kidney tissue paraffin sections are subjected to' H&E "staining," Masson "staining and" Vonkossa "staining, and H was observed with a microscope&E stained section to determine renal tissue injury, Masson stained section to determine renal tissue fibrosis, Vonkossa stained section to determine renal calcium oxalate calculus, and further using Polarized light microscope to observe H&E staining the sections to determine the condition of the kidney crystalline stones.
And (3) test results: the dose effect of the kidney tissue damage of a mouse model after 5 days of glyoxylate intraperitoneal injection is shown in figure 2 (magnification: 200 times), as can be seen from figure 2, H & E staining indicates that the kidney tissue of the mouse model can be seen in sheet-shaped tubular acute injury, which is manifested by that the brush border of the tubular organ falls off, epithelial cells are flat, the tubular cavity expands cystic, and interstitial focal inflammatory cell infiltration, after treatment with different doses of magnesium hydride, the pathological table of the tubular acute injury is indicated to be relieved, and simultaneously, a dose effect relationship exists, the larger the dose of the magnesium hydride is, the lighter the degree of the kidney injury is, the better the effects of the Gly + MH200 and Gly + MH400 groups are, and the treatment effects are not obviously different; the deposition of collagen fibers blue-stained in the interstitial spaces of the kidney of a model mouse is obviously increased by Masson staining, which indicates that the kidney fibrosis is obvious, the kidney fibrosis of a treatment group is relieved, and the effects of Gly + MH200 and Gly + MH400 are more obvious; vonkossa staining shows that a large amount of black crystals are deposited in a renal tubule lumen of a model mouse to block the lumen, black crystal infiltration can be seen in part of interstitium, equal inflammatory cell infiltration is seen in dense crystal positions, the deposition of Gly + MH50 group crystals in a treatment group is not obviously improved, the deposition of Gly + MH100 group crystals is obviously reduced, and the black crystal staining is hardly seen in Gly + MH200 and Gly + MH400 groups; the crystal is in double-folded light under the polarized light of the H & E dyed slice, which shows that the calcium oxalate animal model in the experiment is successfully induced, the formed crystal/calculus component is calcium oxalate, and in a treatment group, the double-folded light crystal is obviously reduced along with the increase of the administration dosage. The results of the above experiments are combined to find that magnesium hydride can significantly reduce the renal tubular injury induced by crystalline calculus, inhibit the formation of renal crystalline calculus and improve the degree of renal fibrosis, and meanwhile, the kidney protection effect is stronger along with the increase of the dosage of magnesium hydride, and when the dosage is greater than 200mg/kg of the body weight of a mouse, the curative effect of the magnesium hydride is not obviously different from that of the dosage of 200mg/kg, so that the dosage of magnesium hydride in the body of the mouse is determined to be 200mg/kg of the body weight of the mouse in subsequent experiments.
(III) the magnesium hydride improves the damage degree of the renal tubules of the mice
The test method comprises the following steps: according to the experimental result of the dose effect of the magnesium hydride, 200mg/kg of magnesium hydride is selectedAnd performing subsequent animal experiments, namely mixing corn oil serving as a solvent with magnesium hydride to form suspension, and reacting with water to hydrolyze to generate hydrogen and magnesium hydroxide after the suspension enters the digestive tract. Con group is prepared by intraperitoneal injection of normal saline with equal dosage for 5 days, Gly group is prepared by intraperitoneal injection of glyoxylate for 5 days, and GH group is prepared by 2 days earlier by MgH 2 Gavage at 200mg/kg, performing intraperitoneal injection of glyoxylate for 5 days, performing intraperitoneal injection of glyoxylate for 4-6 hours twice, and performing intraperitoneal injection of Mg (OH) for 2 days in advance for GOH group 2 The 446mg/kg mouse body weight is used for gastric lavage, the glyoxylate is continuously administered for 5 days after 2 days, and the administration time interval of the two glyoxylate intraperitoneal injections is 4-6 hours; the GV group is administered with corn oil by intragastric administration 2 days earlier, and is administered with glyoxylate intraperitoneal injection for 5 days after 2 days, and the administration time of two glyoxylate intraperitoneal injections is 4-6 hours.
And (3) test results: the results are shown in FIG. 3 (magnification: 200 times), and from H & E, VonKossa, Masson staining of kidney tissue of five groups of mice, it is clear that magnesium hydride group (GH group) improved glyoxylate-induced acute injury of renal tubules of renal mice, and Masson staining suggested that interstitial fibrosis was also somewhat reduced, while GOH group and GV group did not have the above-mentioned effects.
(IV) magnesium hydride improves the renal function level of the crystallized renal injury mice
The test method comprises the following steps: 5 days after the mice are injected with the glyoxylate in the abdominal cavity, collecting a blood sample of the mice, centrifuging to obtain serum, and detecting the serum creatinine and urea nitrogen levels of the mice by using a full-automatic biochemical analyzer.
And (3) test results: as can be seen from FIG. 4, the serum urea nitrogen and creatinine of the mice in the Gly group were both significantly increased as compared with the Con group, and the renal function was decreased after the treatment with magnesium hydride, while the GOH and GV groups did not have the above-mentioned protective effects.
(V) magnesium hydride reduces the level of inflammation in mice with crystallized renal injury
The test method comprises the following steps: 5 days after the mice are injected with the glyoxylate in the abdominal cavity, blood samples of the mice are collected and centrifuged to obtain serum, and the IL-1 beta and TNF-alpha levels of the serum of the mice are detected by an ELISA method.
And (3) test results: the test result is shown in figure 5, the IL-1 beta and TNF-alpha values of serum in the Gly group are increased, the inflammation indexes in the GH group are reduced, the difference has obvious significance, and the magnesium hydride has an improvement effect on the inflammation reaction of the crystallized kidney injury.
(VI) magnesium hydride improves the renal tissue oxidative stress and lipid peroxidation indexes of the mice with the crystallized renal injury
The test method comprises the following steps: the oxidative stress indexes (GSH and NO) and the lipid peroxidation indexes (MDA and 4HNE) of the kidney tissues of the crystallized kidney-injured mice are detected by using an oxidative stress and lipid peroxidation kit and a multifunctional enzyme labeling instrument.
And (3) test results: as shown in FIG. 6, the oxidative stress level and lipid peroxidation level of the body of the mice with the kidney injury are obviously increased, while magnesium hydride can eliminate oxygen free radicals in the body and improve the oxidative stress and lipid peroxidation level of the body.
(VII) magnesium hydride for improving renal tubular cell apoptosis of mice with crystallized renal injury
The test method comprises the following steps: the in situ terminal transferase labelling technique (TdT-mediated DUTP nickend labeling, TUNEL) reflects the fragmentation of nuclear DNA during early apoptosis. Dewaxing and rehydrating the paraffin sections; proteinase K digestion, exposed DNA, 20. mu.g/ml, incubation for 15 min at room temperature, 5 min × 3 washes with PBS; incubation in TUNEL reaction solution at room temperature for 1 hour; DAB color development, 10 minutes, PBS wash 5 minutes x 3 times; hematoxylin counterstain the nucleus; and (3) performing gradient dehydration on alcohol, enabling dimethylbenzene to be transparent, sealing a piece, and observing and photographing under an optical microscope.
And (3) test results: the results are shown in fig. 7, the scattered cell nuclei of the renal tubular epithelial cells of the crystallized renal mouse (indicated by an arrow) are brownish yellow cells, namely the apoptotic cells exist, the renal tubular epithelial cells of the GOH group and the GV group, the renal tubular epithelial cells of the magnesium hydride intervention group have normal morphology, and no apoptotic cells are found.
It should be noted that the embodiments of the present invention have been described in terms of preferred embodiments, and not by way of limitation, and that those skilled in the art can make modifications and variations of the embodiments described above without departing from the spirit of the invention.
Claims (7)
1. The application of magnesium hydride in preparing medicine for preventing and treating renal crystalline lithiasis is provided.
2. The use of magnesium hydride in the preparation of a medicament for the prevention and treatment of renal calculus, as claimed in claim 1, wherein the medicament is a magnesium hydride as the sole active ingredient, or a pharmaceutical composition comprising magnesium hydride.
3. The use of magnesium hydride in the preparation of a medicament for the prevention and treatment of renal calculus, as claimed in claim 1, wherein the magnesium hydride is present in the medicament in an amount of 0.1 to 99% by weight.
4. The use of magnesium hydride in the preparation of a medicament for the prevention and treatment of renal calculus, as claimed in claim 2, wherein said pharmaceutical composition further comprises pharmaceutically acceptable excipients.
5. The use of magnesium hydride in the preparation of a medicament for the prevention and treatment of renal calculus, according to claim 4, wherein the pharmaceutically acceptable excipients are selected from one or more of corn oil, glycerol, propylene glycol, and water.
6. Use of magnesium hydride according to any one of claims 1 to 5 for the preparation of a medicament for the prevention and treatment of renal calculus, wherein said medicament is in the form of an oral preparation or an injectable preparation.
7. An oral preparation for preventing and treating kidney crystal lithiasis is characterized by comprising magnesium hydride and pharmaceutically acceptable auxiliary materials, wherein the pharmaceutically acceptable auxiliary materials are selected from one or more of corn oil, glycerol, propylene glycol and water.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102462694A (en) * | 2010-11-12 | 2012-05-23 | 上海交通大学医学院附属第三人民医院 | Application of hydrogen to preparation of medicament and health-care beverage for preventing kidney stones |
| CN105456288A (en) * | 2015-12-15 | 2016-04-06 | 上海交通大学 | Healthcare pharmaceutical preparation with magnesium supplementation and antioxidation functions and preparation method thereof |
| US20170106032A1 (en) * | 2014-04-02 | 2017-04-20 | Universitat De Les Illes Balears | Combination of a urinary acidifier and a calcium phosphate crystallisation inhibitor for the treatment or prevention of renal lithiasis |
-
2021
- 2021-02-19 CN CN202110195051.4A patent/CN114948989A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102462694A (en) * | 2010-11-12 | 2012-05-23 | 上海交通大学医学院附属第三人民医院 | Application of hydrogen to preparation of medicament and health-care beverage for preventing kidney stones |
| US20170106032A1 (en) * | 2014-04-02 | 2017-04-20 | Universitat De Les Illes Balears | Combination of a urinary acidifier and a calcium phosphate crystallisation inhibitor for the treatment or prevention of renal lithiasis |
| CN105456288A (en) * | 2015-12-15 | 2016-04-06 | 上海交通大学 | Healthcare pharmaceutical preparation with magnesium supplementation and antioxidation functions and preparation method thereof |
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