CN116440271A - Application of miR-19a-3p inhibitor in preparation of neuroprotective drugs - Google Patents
Application of miR-19a-3p inhibitor in preparation of neuroprotective drugs Download PDFInfo
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Abstract
The invention relates to the technical field of neuroprotection medicine preparation, and in particular discloses application of a miR-19a-3p inhibitor in preparation of neuroprotection medicines, wherein the miR-19a-3p inhibitor comprises the following components in percentage by weight: 5'-UCAGUUUUGCAUAGAUUUGCACA-3'. The miR-19a-3p inhibitor can obviously reduce miR-19a-3p expression, reduce damage of rat hippocampal CA1 zone neurons after cerebral ischemia reperfusion, and improve cognitive function of rats after cerebral ischemia reperfusion.
Description
Technical Field
The invention relates to the technical field of preparation of neuroprotective medicines, in particular to application of a miR-19a-3p inhibitor in preparation of neuroprotective medicines.
Background
Cerebral stroke is the second leading cause of death and disability in the world today, of which about 87% are ischemic strokes. Ischemic cerebral apoplexy is a disease which causes neuron damage due to lack of brain tissue blood supply and further causes nerve dysfunction, has five characteristics of high morbidity, high disability rate, high mortality rate, high recurrence rate and high economic burden, and brings heavy burden to life and society of people. The main measure of the current clinical treatment of ischemic cerebral apoplexy is to recover blood supply as soon as possible by thrombolysis and other modes. However, in the process of recovering blood supply after ischemia, the influence of factors such as calcium overload, oxygen free radicals and the like can cause the re-injury of brain tissues, namely, cerebral ischemia reperfusion injury, and seriously influence the prognosis effect of patients. How to effectively alleviate cerebral ischemia reperfusion injury is a problem to be solved. Although a number of exogenous drugs (including radical scavengers, anti-inflammatory molecules, neurotrophic factors, etc.) have been developed to reduce cerebral ischemia reperfusion injury, the clinical use of these drugs has not achieved the desired effect. Therefore, there is a great need in the neuroprotection field to develop new brain cell protective drugs.
In recent years, post-ischemic adaptation, i.e. the one or more relatively brief ischemia reperfusion of a particular tissue during post-cerebral ischemia reperfusion, has proven to be a very effective endogenous brain protection strategy. Post-ischemic adaptation significantly reduces nerve damage following cerebral ischemia reperfusion by interfering with the reperfusion process following cerebral ischemia, continues to significantly improve brain function, but its endogenous neuroprotective mechanisms have not yet been fully elucidated. The intrinsic molecular mechanism of the adaptation protection effect after ischemia is deeply explored, and a safe and effective novel medicine can be provided for brain protection treatment.
microRNA is a non-coding RNA molecule expressed endogenously by an organism, can participate in regulating and controlling a plurality of target genes, can be specifically combined with target gene mRNA through a complete complementation or partial complementation mode, further regulates and controls the expression of the target genes, and can play an important role in pathological processes. The micro RNA or the effector targeting the micro RNA has the characteristics of small molecules, easy administration, long action duration and the like, and the advantages enable the micro RNA or the effector targeting the micro RNA to be more easily converted into potential drugs for clinical treatment. The present invention is directed to the development of neuroprotective agents of this type.
Disclosure of Invention
In order to solve the technical problems, the invention provides application of a miR-19a-3p inhibitor in preparation of neuroprotective medicines, wherein the miR-19a-3p inhibitor comprises the following components: 5'-UCAGUUUUGCAUAGAUUUGCACA-3'.
Further, the miR-19a-3p inhibitor can be used for preparing medicines for improving cognitive functions of rats after cerebral ischemia reperfusion.
Further, the miR-19a-3p inhibitor can be used for preparing medicines for reducing damage of rat neurons after cerebral ischemia reperfusion.
Further, the miR-19a-3p inhibitor can be used for preparing medicines for reducing damage of rat hippocampal CA1 region neurons after cerebral ischemia reperfusion.
Further, the miR-19a-3p inhibitor is used for reducing expression of miR-19a-3 p.
Compared with the prior art, the invention has the beneficial effects that:
1. the miR-19a-3p inhibitor designed by the invention can obviously reduce the on-body expression level of miR-19a-3p, reduce the damage of neurons in the CA1 region of the hippocampus in the ischemia sensitive region, improve the cognitive function of rats and has the potential for preparing neuroprotective medicines.
2. According to the invention, the miR-19a-3p inhibitor is injected 30 minutes after the rat model with cerebral ischemia reperfusion injury is molded, namely, the miR-19a-3p inhibitor is injected 30 minutes after severe 15 minutes whole brain ischemia reperfusion, so that the delayed death of the hippocampal CA1 neurons in an ischemia sensitive area can be reduced, and the cognitive function of the rat can be improved.
3. The invention expands the current cognition of the action of miR-19a-3p in brain injury, provides a potential target point for neuroprotection treatment of ischemic stroke, and is also expected to provide a new candidate marker for clinical diagnosis of ischemic stroke, and the miR-19a-3p inhibitor has potential for preparing neuroprotection treatment medicines.
4. The invention finds that the expression level of the miR-19a-3p in the CA1 region of the rat hippocampus is obviously reduced in a post-ischemia adaptive neuroprotection model, and the post-ischemia adaptation inhibits the expression level of the miR-19a-3p in the CA1 region of the hippocampus after cerebral ischemia reperfusion.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the relative expression level of miR-19a-3p in blood of an acute ischemic stroke group (reference: blood of healthy people, patient: blood of 60-75 years old acute ischemic stroke patient, a P<0.05 represents the comparison of the patient group with the control group, n=8).
FIG. 2 shows the relative expression levels of rat hippocampal CA1 region miR-19a-3p at reperfusion for 6h, 12h and 24h (note: a P<0.05 represents the comparison with the sham group, b P<0.05 represents n=4 compared to the cerebral ischemia reperfusion group).
FIG. 3 shows the effect of post-ischemic adaptation on injured neurons in the CA1 region of the hippocampus of rats after cerebral ischemia reperfusion in the present invention;
wherein, the graph A shows the Nib staining graphs of each group (a-c, hippocampus global graph; d-f, hippocampus CA1 region local enlargement graph; black square indicates local enlargement part);
panel B shows pyramidal neuronal density [ ] for survival in the CA1 region of the hippocampus of rats in each group a P<0.01 represents the comparison with the sham group, b P<0.01 represents n=5 compared to the cerebral ischemia reperfusion group).
FIG. 4 shows the effect of post-ischemic adaptation on rat open field and new object recognition behavior after cerebral ischemia reperfusion in accordance with the present invention;
wherein, panel a represents the body weight of each group of rats (n.s. represents comparative no statistical difference, n=6);
panel B shows the average speed of each group of rats in the open center region (n.s. shows no statistical difference, n=6);
panel C shows residence time of each group of rats in the central region of open field (n.s. shows comparative no statistical difference, n=6);
graph D shows the new object cognitive index of each group of rats a P<0.01， b P<0.05 represents a comparison with the sham group; c P<0.05 represents n=6 compared to the cerebral ischemia reperfusion group).
FIG. 5 shows the effect of miR-19a-3p inhibitors of the invention on rat hippocampal CA1 region injured neurons after cerebral ischemia reperfusion (control: cerebral ischemia reperfusion + control: miR-19a-3p inhibitor: cerebral ischemia reperfusion + miR-19a-3p inhibitor);
wherein, the graph A shows the relative expression level of each group of miR-19a-3p a P<0.01 represents n=4 compared to the control group);
panel B shows Nib staining plots of each group (a-B, hippocampal global plot; c-d, hippocampal CA1 region local enlargement; black boxes indicate local enlargement);
panel C shows pyramidal neuron density [ ] surviving the CA1 region of the hippocampus of each group of rats a P<0.01 represents n=5 compared to the control group).
FIG. 6 shows the effect of miR-19a-3p inhibitors of the invention on rat open field and new-body recognition behavior following cerebral ischemia-reperfusion;
wherein, panel a represents the body weight of each group of rats (n.s. represents comparative no statistical difference, n=6);
panel B shows the average speed of each group of rats in the open center region (n.s. shows no statistical difference, n=6);
panel C shows residence time of each group of rats in the central region of open field (n.s. shows comparative no statistical difference, n=6);
graph D shows the new object cognitive index of each group of rats a P<0.05 represents n=6 compared to the control group).
Detailed Description
The following detailed description of specific embodiments of the invention is, but it should be understood that the invention is not limited to specific embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The experimental methods described in the examples of the present invention are conventional methods unless otherwise specified, and materials, reagents, etc. used in the examples described below are commercially available.
Example 1
Application of miR-19a-3p inhibitor in preparation of neuroprotective drugs
1. Materials and methods
1. Inhibitors of miR-19a-3 p: 5'-UCAGUUUUGCAUAGAUUUGCACA-3' (SEQ ID NO. 1);
miR-19a-3p inhibitors illustrate: the whole chain of the miR-19a-3p inhibitor (namely antagomir of miR-19a-3 p) is subjected to methylation modification, respectively subjected to thio modification at the front 2 bases of the 5' end and the rear 4 bases of the 3' end, and connected with cholesterol modification at the 3' end. Designed by the applicant itself and produced by su Ji Ma gene stock.
2. Experimental animals and main reagents
Adult SD male rats were selected and weighing 200-300 g, provided by Xuzhou university of medical science laboratory animal center. Experimental animal use license: SYXK 2020-0048, all experimental procedures of this study were approved by the Xuzhou university laboratory animal center ethical committee.
The Nib staining solution was purchased from Shanghai Biyun biotechnology Co., ltd; trizol reagent was purchased from Invitrogen corporation, USA; reverse transcription kits and SYBR Green Polymerase Chain Reaction (PCR) kits were purchased from TaKaRa, japan.
The miR-19a-3p primer and the U6 primer are produced by biological engineering (Shanghai) Co., ltd. And the miR-19a-3p inhibitor and the control agent are produced by Suzhou Ji Ma Gene Co., ltd.
2. Experimental method
1. Clinical blood sample collection method
All subjects collected peripheral venous blood using EDTA anticoagulant tubes and stored at-80 ℃. The clinical research project is approved by the medical ethics committee of the affiliated hospital of Xuzhou medical university.
Inclusion criteria: acute ischemic stroke is diagnosed for the first time in the hospital by combining clinical symptoms with imaging CT or MRI, the age range is 60-75 years, and the stroke scale score is 4-15 points through the national institutes of health.
Exclusion criteria: other types or secondary cerebral stroke, such as subarachnoid hemorrhage, brain tumor, and cerebral vascular malformation, etc.; recurrent cerebral apoplexy; kidney and liver system diseases; cancer; acute infectious diseases; inflammation and autoimmune diseases; mental diseases such as depression and schizophrenia.
Healthy crowd selection criteria: the age is matched with the group of patients suffering from acute ischemic cerebral apoplexy, all meet the exclusion standard, and no cerebral apoplexy risk factors exist.
2. Animal experiment grouping and treatment
91 SD rats were randomly divided into 5 groups: a sham operation group, a cerebral ischemia reperfusion group, a post-ischemia adaptation group, a cerebral ischemia reperfusion + contrast agent group and a cerebral ischemia reperfusion + miR-19a-3p inhibitor group.
The brain ischemia reperfusion group and post-ischemic adaptation group rats were 23 each, and the remaining three groups were 15 each. A model of whole brain ischemia reperfusion was established by four arterial ligation (Zhu QJ, kong FS, xu H, et al Tyrosine phosphorylation of GluK2 up-regulates kainate receptor-mediated responses and downstream signaling after brain ischemia). The rat separates the bilateral common carotid artery under anesthesia and electrically coagulates the vertebral artery, and after 24 hours, the bilateral common carotid artery is clamped by an arterial clamp under awake state of the rat, and after 15 minutes, the arterial clamp is removed to restore blood flow supply. The rectal temperature is maintained between 36.5 and 37.5 ℃ during ischemia. Sham rats were given the same surgical treatment, but did not occlude the common carotid artery on both sides. The post-ischemia adaptation group is prepared according to the whole brain ischemia reperfusion model, and ischemia is performed again after 15min of ischemia, 10min of reperfusion, and 3min of reperfusion is resumed. The cerebral ischemia reperfusion + control group was 10. Mu.L of control agent administered at 0.01. Mu. Mol/L to the unilateral ventricle 30min after molding. The cerebral ischemia reperfusion + miR-19a-3p inhibitor group was prepared by administering 0.01. Mu. Mol/L miR-19a-3p inhibitor 10. Mu.L to a unilateral ventricle 30min after molding. Contrast agent: 5'-CAGUACUUUUGUGUAGUACAA-3' (SEQ ID NO. 2), miR-19a-3p inhibitor: 5'-UCAGUUUUGCAUAGAUUUGCACA-3'.
3. qRT-PCR method for detecting clinical blood sample and expression level of rat hippocampal CA1 region miR-19a-3p
Blood samples and brain tissue total RNA were extracted with Trizol reagent and then reverse transcribed into cDNA using reverse transcription kit. The obtained cDNA was used as a template to prepare a PCR reaction solution according to SYBR Premix Ex Taq II, and qRT-PCR was performed on a fluorescent quantitative PCR apparatus. U6 as reference geneAdopts 2 -ΔΔCT The relative expression level of miR-19a-3p was calculated by the method.
Primer information is as follows:
miR-19a-3p reverse transcription primer (shown in SEQ ID NO. 3): 5'-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTCAGTTT-3';
miR-19a-3p upstream primer (shown in SEQ ID NO. 4): 5'-CTGGAGTGTGCAAATCTATGCA-3';
miR-19a-3p downstream primer (shown in SEQ ID NO. 5): 5'-GTGCAGGGTCCGAGGT-3';
u6 reverse transcription primer (shown as SEQ ID NO. 6): 5'-AAAATATGGAACGCTTCACGAATTTG-3';
u6 upstream primer (as shown in SEQ ID NO. 7): 5'-CTCGCTTCGGCAGCACATATACT-3';
u6 downstream primer (as shown in SEQ ID NO. 8): 5'-ACGCTTCACGAATTTGCGTGTC-3'.
4. Nib staining method for observing morphology and survival neuron density of rat hippocampal CA1 region neurons
(1) Tissue sample preparation
At cerebral ischemia reperfusion for 5d, rats were anesthetized and fixed on dissecting plates in supine, the chest was opened to expose the heart, a perfusion needle was inserted into the aorta from the left apex of the heart through the left ventricle, a small opening was cut in the right auricle of the animal heart, the animals were rapidly rinsed with approximately 300ml of physiological saline, 200ml of buffer solution containing 4% paraformaldehyde was refilled, the rats were broken after the end of the perfusion, and the brain tissue was removed and fixed in 4% paraformaldehyde solution.
(2) Paraffin section preparation and Nib staining
The brain tissue is fixed in 4% paraformaldehyde solution for 12-18 h, washed by running water for 0.5-1 h, dehydrated by 60%, 70%, 80%, 90%, 95% and 100% gradient ethanol, transparent for xylene, and embedded after wax dipping at 60 ℃. The embedded wax block is sliced by a paraffin slicer (thickness is 6 μm), and is pasted on a glass slide, spread for 2h at 63 ℃ and stored at normal temperature. Paraffin sections were dewaxed with xylene, immersed in 100%, 90%, 70% gradient ethanol and ultrapure water, stained with nikov stain for 3min to 10min, water washed to terminate, dehydrated with 95% ethanol, transparent with xylene, and after neutral resin sealing, the cell morphology was observed under a microscope and counted. The number of normal morphology pyramidal neurons in the rat hippocampal CA1 region over a length of 1mm was counted as the neuron density.
5. Behavioural experimental method
(1) Open field experiment
The observation box is divided into a central area and four sides. The rats were removed from the feeder cages and placed quickly in the center of the observation box, allowing free movement in the box for 5min. The activity of the rats in the box was automatically recorded by using behavioural analysis software, and the movement speed and the residence time in the central zone were analyzed. After each rat experiment is finished, the observation box is wiped clean by 75% ethanol, so that the subsequent rat experiment is prevented from being interfered.
(2) New object identification
The new object recognition experiment is divided into three stages: the adaptation period, familiarity period and testing period are completed in 3 days. During the first day adaptation period, rats were placed in the incubator for 5min of free movement. In the next familiarity period, two identical and symmetrical objects are placed in the box, the rats are placed from the middle point of the non-object area, the rats are allowed to freely explore for 5min, after 1h interval, the rats are placed from the same position, and the rats are allowed to freely explore for 5min again. After the familiarity period is ended for 24 hours, the test period is started, one of the old objects is replaced by a new object with different colors and shapes, the rat is free to explore for 5 minutes, and the exploration time of the rat for the new object and the old object is recorded respectively. The placement positions of the new object and the old object are required to be sequentially rotated. In different experimental stages and between different rats, the observation box and the object are all wiped by 75% ethanol, so that the interference to the subsequent animal behaviors is avoided. The exploration activities are defined as: the nose of the rat is directly directed to the object or the front claw and the like to directly contact the object within the range of less than or equal to 2 cm. New object recognition index = new object exploration time/(new object exploration time + old object exploration time) ×100%.
6. Statistical method
The comparison between two groups adopts t test, the single factor comparison between a plurality of experimental groups adopts single factor analysis of variance (One-way ANOVA), and P <0.05 is statistically significant. Blood sample data and animal behavioral data are expressed as Mean ± standard error (Mean ± SEM) and other metering data are expressed as Mean ± standard deviation (Mean ± SD).
3. Results
1. Upregulation of miR-19a-3p expression in blood of acute ischemic stroke patient
In the invention, patients suffering from acute ischemic cerebral apoplexy are admitted and peripheral blood is collected within 24 hours of onset, wherein 4 cases of acute aortic infarction and 4 cases of acute lacunar infarction are respectively treated. As shown in fig. 1, the expression level of miR-19a-3P in blood of acute ischemic stroke group was significantly higher than that of control group (P < 0.05).
2. Post-ischemic adaptation inhibition of brain ischemia reperfusion-induced up-regulation of expression of rat hippocampal CA1 region miR-19a-3p
The relative expression of miR-19a-3p in the CA1 region of the hippocampus of rats in each of the groups of 6h, 12h and 24h of cerebral ischemia reperfusion was detected by qRT-PCR, and the result is shown in FIG. 2. No significant differences were seen in miR-19a-3P expression levels between the sham-operated, cerebral ischemia-reperfusion and post-ischemic adaptation groups (P > 0.05) at reperfusion for 6h and 12 h. At 24h reperfusion, the brain ischemia reperfusion group had significantly increased miR-19a-3P expression level (P < 0.05) compared to the sham surgery group, while post-ischemia adaptation group had significantly decreased miR-19a-3P expression level (P < 0.05); the post-ischemic adapted group also had significantly reduced miR-19a-3P expression levels (P < 0.05) compared to the cerebral ischemia reperfusion group.
3. Adaptation after ischemia to reduce damage to rat hippocampal CA1 region neurons after cerebral ischemia reperfusion
The experiments were divided into sham surgery groups, cerebral ischemia reperfusion groups and post-ischemic adaptation groups. As shown in fig. 3 (a), in cerebral ischemia reperfusion for 5 days, the hippocampus structure of the sham rat is complete and clear, the cell morphology of the CA1 zone neurons is regular, the nuclei are clear, and 2-3 layers of cone neurons are orderly and closely arranged; the neurons in the CA1 region of the hippocampus of the rat in the cerebral ischemia reperfusion group die in large quantity, the cell structure is destroyed, and residual fragments of the dead neurons can be seen; the arrangement of neurons in the CA1 region of the hippocampus of the adaptation group rats after ischemia is slightly relaxed, and the scattered neurons can die. As shown in fig. 3 (B), the density of pyramidal neurons surviving in the CA1 region was significantly reduced (P < 0.01) in both the cerebral ischemia reperfusion group and post-ischemic adaptation group rats compared to the sham surgery group; compared to the cerebral ischemia reperfusion group, the post-ischemic adapted group had significantly increased pyramidal neuronal density surviving the CA1 region of the rat hippocampus (P < 0.01).
4. Adaptation after ischemia to repair cognitive function in rats after cerebral ischemia reperfusion
At reperfusion 6d, rats of the sham operation group, the cerebral ischemia reperfusion group and the post-ischemia adaptation group were placed in an open field experimental observation box, the movement speed and the residence time of the rats in the central area of the open field were recorded, and the body weight was weighed. As shown in fig. 4 (a-C), the body weight, movement speed in the central region of open field and residence time were not significantly different for each group of rats, indicating that the autonomous movement ability and anxiety degree of each group of rats were comparable. As shown in fig. 4 (D), the new object recognition experiments are performed at reperfusion of 7D to 9D, and compared with the sham operation group, the cognitive indexes of rats in the cerebral ischemia reperfusion group and the post-ischemia adaptation group are significantly reduced (P <0.01, P < 0.05), which indicates that the cognitive function is impaired; compared to the cerebral ischemia reperfusion group, post-ischemic adaptation group rats had significantly increased cognitive index (P < 0.05) (see table 1 for specific data).
Table 1 influence of post-ischemic adaptation on rat open field and new object recognition behavior after cerebral ischemia reperfusion (mean±sem, n=6)
Note that: a P<0.01， b P<0.05 represents a comparison with the sham group; c P<0.05 shows comparison with the cerebral ischemia reperfusion group
5. Inhibiting miR-19a-3p reduces damage to rat hippocampal CA1 region neurons after cerebral ischemia reperfusion
The experiments were divided into a cerebral ischemia reperfusion + control group and a cerebral ischemia reperfusion + miR-19a-3p inhibitor group. As shown in FIG. 5 (A), the relative expression level of miR-19a-3p in the CA1 region of the hippocampus was significantly reduced in the rat in the miR-19a-3p inhibitor group compared with the control group at 24h of reperfusion. As shown in fig. 5 (B), the number of neurons in the CA1 region of the hippocampus of the rats in the control group was greatly reduced at 5d reperfusion, and the cell morphology was incomplete, and a large amount of dead cell debris was seen; while the neurons in the CA1 region of the hippocampus of the inhibitor group are densely arranged, the neuron structure is clear, and residual fragments of a small amount of dead neurons can be seen. As shown in fig. 5 (C), the density of pyramidal neurons surviving the CA1 region of the hippocampus was significantly increased in the miR-19a-3P inhibitor group compared to the control group (P < 0.01).
6. Inhibiting miR-19a-3p improves cognitive function of rat after cerebral ischemia reperfusion
The experiments were divided into a cerebral ischemia reperfusion + control group and a cerebral ischemia reperfusion + miR-19a-3p inhibitor group. As shown in fig. 6 (a-C), the miR-19a-3p inhibitor group rats had no significant difference in weight, movement speed in the central region of open field, and residence time compared to the control group, indicating that the autonomous movement ability and anxiety degree of the two groups of rats were comparable. As shown in fig. 6 (D), in the new object recognition experiments performed later, the cognitive index of the miR-19a-3P inhibitor group rats was significantly increased (P < 0.05) compared to the control group, suggesting an improvement in cognitive ability of the rats (see table 2 for specific data).
Table 2 Effect of miR-19a-3p inhibitors on rat open field and New object recognition behavior following cerebral ischemia reperfusion (mean+ -SEM, n=6)
Note that: control group: cerebral ischemia reperfusion + control group; miR-19a-3p inhibitor group: cerebral ischemia reperfusion + miR-19a-3p inhibitor group; a P<0.05 represents a comparison with the control group.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. It is therefore intended that the following claims be interpreted as including the preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.
Claims (5)
1. The application of the miR-19a-3p inhibitor in the preparation of neuroprotective medicines is characterized in that the miR-19a-3p inhibitor is as follows: 5'-UCAGUUUUGCAUAGAUUUGCACA-3'.
2. The use of claim 1, wherein the miR-19a-3p inhibitor is capable of being used in the manufacture of a medicament for improving cognitive function in a rat following cerebral ischemia reperfusion.
3. The use of claim 2, wherein the miR-19a-3p inhibitor is capable of being used in the manufacture of a medicament for reducing injury to rat neurons following cerebral ischemia reperfusion.
4. The use of claim 3, wherein the miR-19a-3p inhibitor is capable of being used in the manufacture of a medicament for alleviating damage to rat hippocampal CA1 area neurons following cerebral ischemia reperfusion.
5. The use of claim 4, wherein the miR-19a-3p inhibitor is for reducing expression of miR-19a-3 p.
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