Application of LncRNA NEAT1 as diagnostic substance for Parkinson's disease

Abstract

The invention relates to an application of LncRNA NEAT1 as a diagnostic substance of Parkinson's disease. Specifically, the LncRNA NEAT1 is used as a marker for diagnosing the Parkinson's disease so as to further diagnose whether the structure and the function of a midbrain nigra region are normal or not and whether the midbrain nigra region suffers from the Parkinson's disease or not, and the LncRNA NEAT1 can be used for developing a diagnostic agent or a kit related to the Parkinson's disease. The invention finds that the LncRNA NEAT1 can be used as a marker for diagnosing the Parkinson's disease for the first time.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of LncRNA NEAT1 as a diagnostic substance for Parkinson's disease. Background Art

[0002] Parkinson's Disease (PD) is a neurodegenerative disease with resting tremor, bradykinesia, muscle rigidity and postural gait disorders as the main clinical manifestations. The main pathological characteristics are the degeneration and death of dopaminergic neurons in the substantia nigra of the midbrain, and the formation of Lewy bodies composed of α-synuclein (α-syn) in the remaining dopaminergic neurons. PD is not exclusive to middle-aged and elderly people. In recent years, the incidence has shown a trend of younger age. With the aging of the population, China may become the world's largest country with Parkinson's disease. In addition, PD is a chronic disease, and most patients have a course of more than ten years and require lifelong treatment. Because the symptoms slowly worsen, there is currently no way to completely cure it. Therefore, in-depth research on the pathogenesis of PD plays a very important role in the development of drugs for the prevention, diagnosis and treatment of Parkinson's disease. It is also a social and biological problem that needs to be solved urgently to achieve healthy aging.

[0003] Pathologically, Parkinson's disease is defined as the loss of dopaminergic neurons in the substantia nigra, accompanied by abnormal deposition of α-synuclein. The clinical manifestations are mainly divided into motor symptoms and non-motor symptoms. Motor symptoms mainly include resting tremor, rigidity, bradykinesia, abnormal posture and gait, etc. Non-motor symptoms mainly include cognitive impairment, intractable constipation, sleep disorders and autonomic dysfunction. At present, the cause of Parkinson's disease has not been clearly defined. Most studies have shown that age, environmental factors, epigenetic regulation, and the interaction of multiple factors are related to the onset of Parkinson's disease. A meta-analysis screened out 11 environmental factors that significantly changed the risk of Parkinson's disease and found that factors that increased the risk included pesticide exposure, head injury, rural life, beta-blocker use, agricultural occupations and drinking well water, while environmental factors associated with reduced risk included drinking coffee, using nonsteroidal anti-inflammatory drugs, using calcium channel blockers and drinking alcohol. Other studies have confirmed that pesticides such as herbicides and insecticides, as well as metal ions such as iron, manganese, and copper, can significantly increase the incidence of Parkinson's disease. In addition, although the proportion of sporadic Parkinson's disease is much higher than that of hereditary Parkinson's disease, genetic factors are still one of the factors that cannot be ignored. At present, more than 19 pathogenic genes have been found to be associated with Parkinson's disease, including SNCA (α-synuclein), PARKIN (Parkin), PARK7 (protein deglycosylation enzyme DJ-1), UCHL1, PINK1 (PTEN-induced kinase I), LRRK2 (leucine-rich repeat kinase), ATP13A2 (ATPase type 13A), PARK10, PARK12, PARK16, EIF4G1, DNAJC6, CHCHD2, and MAPT. The mainstream view is that the interaction of multiple pathologies such as abnormal aggregation of α-synuclein, mitochondrial dysfunction, inflammatory response, oxidative stress damage, cell autophagy, and cell apoptosis leads to the onset and progression of Parkinson's disease. Although research results on various factors related to the onset of Parkinson's disease have been reported, finding stable biogenetic indicators that are easy to measure to predict, diagnose and mark the occurrence and development of the disease is currently a focus of clinical research on Parkinson's disease.

[0004] In the post-genomic era, more and more studies have found that higher organisms, especially mammals, produce thousands of non-coding RNAs (ncRNAs), which are regulators of many important biological processes, including nervous system development, neurodegenerative diseases, and reproductive system diseases. Compared with protein-coding genes, ncRNAs show strong specificity in spatiotemporal expression. Studies have found that in humans and mice, the number and types of ncRNAs are highly enriched in the reproductive and nervous systems, which further suggests that ncRNAs may play an important role in the nervous and reproductive systems. Among various ncRNAs, long non-coding RNAs (lncRNAs) are a class with diverse functional mechanisms. It refers to a group of non-coding RNAs with a transcription length of more than 200 nucleotides in eukaryotic cells, which have tissue specificity and conditional expression specificity. Previous studies have shown that lncRNAs are enriched in the central nervous system (CNS), showing a complex spatiotemporal expression profile, and participate in the regulation of brain development in various ways. In recent years, more and more studies have suggested that the dysregulation or mutation of lncRNA gene loci is closely related to a variety of central nervous system (CNS)-related neurodegenerative diseases (NDDs), such as Alzheimer's disease, Parkinson's disease, Huntington's disease (HD) and amyotrophic lateral sclerosis (ALS). Although a lot of progress has been made in the study of pathogenic genes in the occurrence of neurodegenerative diseases, the main cause of the disease is still not fully resolved. Non-coding RNA, especially lncRNA, can regulate gene expression in multiple ways at multiple levels such as transcription or post-transcription, thereby playing a role in regulating important life activities, providing a new perspective for exploring the pathogenesis of a variety of CNS-related neurodegenerative diseases, and also providing new strategies and methods for the treatment of these diseases.

[0005] LncRNAs have a strong intrinsic connection with CNS function, and many studies have also suggested that they play an important role in CNS diseases [1-2]. Studies have found that lncRNAs such as MALAT1, NEAT1 and SNHG1 are involved in the pathogenesis of PD. Boros et al. tested the peripheral blood of 43 PD patients and 36 controls and found that NEAT1 expression in PD patients' blood samples was significantly upregulated [3-4]. When gender-matched, there was no significant difference in NEAT1 expression in male patients, while it was significantly upregulated in female patients; and NEAT1 expression was most significantly upregulated in patients with a longer duration of disease [5]. Studies have also found that the expression of NEAT1 and α-synuclein in the substantia nigra dopaminergic neurons of PD model mice was positively correlated [6]. In summary, reports have shown that LncRNA NEAT1 expression is significantly upregulated during the onset of PD. However, the specific relationship between NEAT1 and PD is still unknown. Therefore, it is necessary to study the specific relationship between lncRNA NEAT1 and Parkinson's disease, which can provide a reliable basis for the clinical prevention, diagnosis and future effective treatment of Parkinson's disease. Summary of the invention

[0006] The purpose of the present invention is to study the specific relationship between lncRNA NEAT1 and Parkinson's disease, so as to provide a marker (lncRNA NEAT1) that can be used as a diagnosis of Parkinson's disease. Specifically, the present invention uses immunofluorescence staining technology to verify the relationship between the expression level of lncRNA NEAT1 and the pathological characteristics related to Parkinson's disease - the loss of dopaminergic neurons in the substantia nigra region of the midbrain, and believes that this lncRNA NEAT1 can be used as a biomarker for the diagnosis and treatment of Parkinson's disease.

[0007] In order to solve the problem of insufficient existing prevention and diagnosis technology for Parkinson's disease, the inventors conducted in-depth research. As a result, by using tyrosine hydroxylase (TH) staining to mark the dopamine neurons in the substantia nigra of the midbrain, it was found for the first time that the PD mouse model was induced by MPTP in wild-type control mice, and the number of TH-positive cells in the midbrain region of the mice was significantly reduced, while in lncRNA NEAT1 knockout mice, the number of TH-positive cells in the midbrain region of the mice was significantly increased. It can be seen that lncRNA NEAT1 is closely related to the pathological feature related to Parkinson's disease-the loss of dopaminergic neurons in the substantia nigra of the midbrain. According to the above results, it is proposed for the first time to use lncRNA NEAT1 as a marker for the diagnosis of Parkinson's disease, so as to further diagnose whether the structure and function of the substantia nigra region of the midbrain are normal and whether Parkinson's disease is suffered.

[0008] Specifically, the present invention relates to using lncRNA NEAT1 as a marker for diagnosing Parkinson's disease, thereby further diagnosing whether the structure and function of the substantia nigra region of the midbrain are normal and whether the patient suffers from Parkinson's disease, and can be used to develop diagnostic agents or kits related to Parkinson's disease.

[0009] definition

[0010] Tyrosine hydroxylase (TH): It is a monooxygenase, which is the rate-limiting enzyme that catalyzes the first step of a series of reactions in which organisms synthesize L-dopamine (DA). The body uses L-tyrosine to generate L-dopa under the catalysis of tyrosine hydroxylase, and L-dopa is decarboxylated under the catalysis of aromatic decarboxylase to finally generate L-dopamine. Due to the important role of tyrosine hydroxylase in dopamine synthesis, the loss of its function or insufficient expression directly affects the synthesis and secretion of dopamine. Dopamine is an important neurotransmitter. The inability of dopaminergic neurons to synthesize dopamine or insufficient secretion can lead to Parkinson's disease.

[0011] MPTP: The full name is 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. It is a neurotoxin that selectively damages dopamine neurons in the substantia nigra and striatum system in humans and primates, reduces dopamine and its metabolites, and causes typical Parkinson's-like symptoms.

[0012] Substantia nigra in the midbrain: The substantia nigra is found throughout the midbrain, and the substantia nigra cells are rich in melanin, which is the main nucleus for synthesizing dopamine in the brain. Under normal physiological conditions, the substantia nigra is an important center for regulating movement. The apoptosis of dopamine neurons in the compact part of the substantia nigra can lead to Parkinson's disease.

[0013] Rotarod test: It is used to evaluate the ability of animals to maintain balance on a rotating rod. As the rotation speed increases, it becomes more difficult for the experimental animals to maintain balance, thereby detecting the motor function of rodents.

[0014] Beneficial Effects

[0015] By detecting the expression of lncRNA NEAT1 in different tissues and organs of mice, it was found that lncRNA NEAT1 was mainly enriched in the spleen and lung tissues of young and old mice, while the expression level in the central nervous system was low. However, some studies have found that shutting down NEAT1 in old animals will enhance memory, and increasing NEAT1 levels in young animals will reduce their memory, so we focus on the function and mechanism of lncRNA NEAT1 in brain tissue. lncRNA NEAT1 homozygous knockout mice and wild-type control mice in the same litter were screened from the existing lncRNA NEAT1 heterozygous knockout mice through genotype identification. Then, these two mice were injected with MPTP drugs to construct PD mouse models, and the rotarod test and novel object recognition test were used to verify the successful construction of PD mouse models. Subsequently, tyrosine hydroxylase (TH) staining was used to mark dopaminergic neurons in the substantia nigra of the midbrain, and it was found that MPTP induction in wild-type control mice caused a significant decrease in the number of TH-positive cells in the substantia nigra of the midbrain of mice. When lncRNA NEAT1 knockout mice were induced with MPTP, the number of TH-positive cells in the substantia nigra of the midbrain of the mice increased significantly, reflecting that under the stimulation of MPTP drugs, the loss of lncRNA NEAT1 is beneficial to the development of dopamine neurons in the substantia nigra of the midbrain. The above results show that lncRNA NEAT1 is closely related to the number of substantia nigra neurons in the midbrain of mice, revealing that lncRNA NEAT1 may play an important role in the development and function of substantia nigra neurons in the midbrain of mice. It can be seen that lncRNA NEAT1 is closely related to the pathological characteristics related to Parkinson's disease-the loss of dopamine neurons in the substantia nigra of the midbrain. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The results of the expression level detection of lncRNA NEAT1 are shown. A: RT-qPCR detection of the expression level of lncRNA NEAT1 in 7 organs (heart, liver, spleen, lung, kidney, testis, epididymis) of young and old mice (n = 3). The data results are expressed as mean ± standard deviation, and the statistical significance is expressed as * P < 0.05, ** P < 0.01, *** P < 0.001. B: RT-qPCR detection of the relative expression level of lncRNA NEAT1 transcripts in the olfactory bulb, cerebral cortex, hippocampus, hypothalamus, cerebellum and spinal cord of adult male mice (n = 3). Using t-test, the data results are expressed as mean ± standard deviation, and the statistical significance is expressed as * P < 0.05, ** P < 0.01, *** P < 0.001.

[0017] Figure 2The construction and identification of lncRNA NEAT1 systemic knockout mice are shown. A: Schematic diagram of lncRNA NEAT1 systemic knockout mice constructed using the CRIPSR / Cas9 system. B: Mouse genotype identification primer information and PCR target band size. C and D: Mouse genotype identification result display diagram.

[0018] Figure 3 The experimental flow chart of the present invention is shown. In adult mice, MPTP was continuously injected intraperitoneally for 7 days. Behavioral tests such as rotarod and novel object recognition were completed within 2 days.

[0019] Figure 4 The behavioral test results of PD model mice are shown. A: Rotarod test pattern diagram. BD: The changes in motor coordination ability of PD model mice were identified by rotarod test (n = 5). The t-test was used, and the data results were expressed as mean ± standard deviation, and the statistical significance was expressed as * P < 0.05, ** P < 0.01, *** P < 0.001. EF: The changes in learning and memory function of PD model mice were identified by novel object recognition experiment (n = 5). The t-test was used, and the data results were expressed as mean ± standard deviation, and the statistical significance was expressed as * P < 0.05, ** P < 0.01, *** P < 0.001.

[0020] Figure 5 The results of the detection of the number of TH-positive neurons are shown. A: Immunofluorescence staining was used to evaluate the number of tyrosine hydroxylase (TH)-positive neurons in the substantia nigra of the midbrain (n = 5). The scale bar is 1 mm. B: The number of HT-positive neurons was quantitatively counted using ImageJ software (n = 5). The data results are expressed as mean ± standard deviation, and the statistical significance is expressed as P value. DETAILED DESCRIPTION

[0021] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings.

[0022] Example 1 Detection of expression level of LncRNA NEAT1

[0023] In order to study whether lncRNA NEAT1 (lncRNA NEAT1 has two isoforms, NEAT1_1 and NEAT1_2, NEAT1_1 has a total length of 3190bp, from NCBI, NCBI reference sequence: NR_003513.3; NEAT1_2 has a total length of 20771bp nucleotide sequence, from NCBI, NCBI reference sequence: NR_131212.1) is related to the development of dopamine neurons in the substantia nigra of the midbrain, we plan to use lncRNA NEAT1 knockout mice donated by Professor Wu Mian's research group at the University of Science and Technology of China to verify the above idea. First, the distribution of lncRNA NEAT1 in young and old mice was confirmed, and the expression level and tissue specificity of lncRNA NEAT1 in different organs and brain partitions of 3-month-old and 18-month-old C57 / BL6 wild-type male mice were detected by RT-qPCR.

[0024] The experiment was carried out according to the literature reports, and TRIzol reagent was used to extract brain tissue RNA. Reverse transcription was performed according to the instructions of the reverse transcription kit (Vazyme, R212-01 / 02). The real-time fluorescence quantitative PCR experiment was performed on the Bio-Rad CFX96 qPCR system according to the real-time fluorescence quantitative PCR reagent provided by Novozymes (Vazyme, Q111-02-AA). The gene expression level was calculated relative to the internal reference gene GAPDH. The primer sequence of the fluorescence real-time quantitative PCR of mRNA was synthesized by Anhui General Biotechnology Co., Ltd., and the specific sequence is shown in Table 1.

[0025] Table 1: Primer sequences for real-time fluorescence quantitative PCR of mRNA

[0026]

[0027] The results showed that the expression level of lncRNA NEAT1 in the central nervous system was low ( Figure 1 A in the figure), but was mainly enriched in the spleen and lung tissues of young and old mice ( Figure 1 B in the figure). The expression of lncRNA NEAT1 in the central nervous system of young and old mice does not show much difference. Combined with the fact that shutting down NEAT1 in old animals enhances memory, and increasing NEAT1 levels in young animals reduces their memory, the present invention focuses on the functional mechanism of lncRNA NEAT1 in brain tissue, especially in the substantia nigra of the midbrain, which is closely related to Parkinson's disease.

[0028] Example 2 Construction and screening of LncRNA NEAT1 knockout mouse model

[0029] To further explore the function of lncRNA NEAT1 in mice, the existing F1 generation mice carrying heterozygous (HT) alleles were co-bred and mated to produce F2 generation mice. Figure 2 B) and PCR amplification for genotyping of offspring mice ( Figure 2 C and D in the figure), LncRNANEAT1 homozygous knockout mice (KO) and wild-type control mice (WT) of the same litter were preliminarily screened. The systemic genotype of lncRNA NEAT1 homozygous knockout mice was consistent and the gene inheritance was stable during reproduction. In the study, it can be used for subsequent research on the function of lncRNA NEAT1 in the development of substantia nigra neurons in the mouse midbrain.

[0030] Example 3 Construction and identification strategy of PD model mice

[0031] In order to further explore and verify the functional mechanism of lncRNA NEAT1 in the PD mouse model, we first need to determine the modeling conditions and use MPTP drugs to intraperitoneally inject mice for one week to induce an acute PD mouse model.

[0032] The specific method for constructing the PD model was to randomly divide 20 male NEAT1 knockout mice (9-10 weeks old) and 20 male wild-type control mice (9-10 weeks old) into PD group and control group. The mice in the PD group were intraperitoneally injected with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP, Sigma Aldrich M0896, 25 mg / kg) once a day, and the mice in the control group were injected with the same volume of sterile saline solution (0.9%) as MPTP hydrochloride. After 7 days of continuous injection, all mice were killed to obtain brain tissue. Subsequently, the brain tissue was stored at −80°C for subsequent experiments.

[0033] To further determine whether this model successfully simulates the clinical symptoms of PD, animal behavioral tests such as novel object recognition and rotarod were used to test whether the cognitive and motor abilities of mice were impaired ( Figure 3 In addition, the mouse midbrain tissue was collected and frozen into sections, and immunofluorescence staining was used to identify the modeling effect.

[0034] Example 4 MPTP drug-induced improvement in learning ability of NEAT1 knockout mice

[0035] During the intraperitoneal injection, the MPTP drug dosage was strictly prepared according to the daily mouse body weight. After the MPTP drug injection, the mice were immediately transferred to the behavioral platform. The novel object recognition experiment is one of the basic experiments used to evaluate the learning and memory ability of mice; the rotating rod experiment and the balance beam experiment are currently one of the effective behavioral methods used to evaluate the motor coordination ability of rodents. Therefore, the model mice were subjected to novel object recognition and rotating rod experiments. The specific experimental methods of the novel object experiment are as follows:

[0036] Build a new object recognition device: The device includes a square black box (40cm×40cm×30cm), video equipment, animal behavior analysis software (The ANY-maze video-tracking system, Stoelting, Wood Dale, USA), and Lego blocks (assembled into three objects, two of which are identical, and the other object is different from the other two objects in color and shape). 2 hours before the experiment, place the mice around the new object recognition device to adapt to the experimental environment; place two identical blocks on one side of the black box, one on the left and one on the right, and gently grab the mouse's tail and put it in the middle of the training field so that the training mouse is at the same distance from the two objects. After the mouse is placed in the device to adapt for 10 minutes, start the video recording device to record the whole process of the mouse exploring the new object. The experimenter needs to stay away from the experimental equipment to avoid interference. The mouse freely explores two identical objects in the field for 3 minutes; on the second day of the formal experiment, the object on the side where the mouse spends less time exploring is replaced with a new object (different in shape and color). The mouse freely explores the two objects for 3 minutes, and the whole process of the mouse exploring the new object is recorded. (The experimental device must be wiped with 75% ethanol between each experiment to eliminate the mouse odor). The mouse cognitive index (Discrimination Index) is calculated using animal behavior analysis software. This index reflects the mouse's preference for new objects and further detects the mouse's cognitive ability.

[0037] The specific experimental method of the rotarod test is as follows:

[0038] Before training, mice were placed in the behavioral laboratory at least 30 minutes in advance to acclimate to the environment. The mouse rotarod fatigue apparatus (XR-6C, Shanghai Xinruan Information Technology Co., Ltd.) was turned on, and mice were placed on the rotating rod apparatus and accelerated to 4-40 rpm for 5 minutes. Most mice fell before 40 rpm. Mice were trained 3 times a day, with a 20-minute interval between each time, for two consecutive days. The rotating rod was cleaned between each test. The waiting time to fall and the total running distance were automatically recorded.

[0039] like Figure 4As shown in the AD results, in WT mice, the distance run on the rotarod was significantly reduced in mice injected with MPTP compared with the control group; in KO mice, the distance run on the rotarod was significantly increased in mice injected with MPTP compared with the control group.

[0040] like Figure 4 As shown in the F results, in WT mice, the recognition index of mice injected with MPTP was significantly decreased compared with the control group; in KO mice, the recognition index of mice injected with MPTP was significantly increased compared with the control group.

[0041] Both experiments found that wild-type (WT) mice induced by MPTP had motor impairment and significantly reduced learning and memory abilities. However, when LncRNA NEAT1 knockout mice were induced by MPTP, their learning and memory abilities were significantly improved despite motor impairment, suggesting that lncRNA NEAT1 may play a certain feedback regulatory role in the movement and learning and memory of mice.

[0042] Example 5 MPTP drug-induced increase in substantia nigra neurons in NEAT1 knockout mice

[0043] In order to further determine whether LncRNA NEAT1 has a significant effect on midbrain neurons in the PD mouse model, according to the drug dose and injection method modeled in the previous step, MPTP was given to lncRNA NEAT1 knockout mice to induce external stimulation, and tyrosine hydroxylase (TH) fluorescence staining was used to mark midbrain substantia nigra dopamine neurons. The specific experimental methods are as follows:

[0044] The tissue sections were taken out in advance, placed in a humidified box, and dried at room temperature for 1 hour; after washing three times, 0.1% Triton X-100 was used to permeabilize for 45 minutes; 3% BSA-PBS solution was used to block at room temperature for 1 hour; antibodies (primary antibody, THPolyclonal antibody, Proteintech, 25859-1) were added and incubated overnight at 4°C; in the dark, fluorescent antibodies (secondary antibodies, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), Abcam, ab150077) were added and incubated at room temperature for 1 hour; Hoechst 33342 (Sigma, B2261) was used to stain the cell nucleus for 2 minutes; PBS was rinsed 3 times, 5 minutes each time, and 10 μL antifade agent (Invitrogen, P36930) was added for sealing and microscopic observation.

[0045] like Figure 5As shown in the AB results, in WT mice, the number of TH-positive cells in mice injected with MPTP was significantly reduced compared with the control group, down 33.83%; in KO mice, the number of TH-positive cells in mice injected with MPTP was significantly increased compared with the control group, up 33.01%.

[0046] This experiment found that MPTP induction in lncRNA NEAT1 knockout mice can cause a significant increase in the number of TH-positive cells in mice. The experimental data further showed that under MPTP drug stimulation, the loss of lncRNA NEAT1 is conducive to the development of dopamine neurons in the substantia nigra of the midbrain, suggesting that lncRNA NEAT1 may play a certain feedback regulatory role in the substantia nigra tissue. This experimental result shows that lncRNA NEAT1 is closely related to the development and function of substantia nigra neurons in the midbrain of mice, revealing that lncRNA NEAT1 plays an important role in the development and function of substantia nigra neurons in the midbrain of mice.

[0047] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[0048] References

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[0051] [3] I. Manna, A. Quattrone, S. De Benedittis, E. Iaccino, A.Quattrone, Roles of Non-Coding RNAs as Novel Diagnostic Biomarkers inParkinson's Disease, J Parkinsons Dis, 11 (2021) 1475-1489.

[0052] [4] M. Majidinia, A. Mihanfar, R. Rahbarghazi, A. Nourazarian, B.Bagca, C.B. Avci, The roles of non-coding RNAs in Parkinson's disease, MolBiol Rep, 43 (2016) 1193-1204.

[0053] [5] F.A. Boros, R. Maszlag-Torok, L. Vecsei, P. Klivenyi, Increasedlevel of NEAT1 long non-coding RNA is detectable in peripheral blood cells ofpatients with Parkinson's disease, Brain Res, 1730 (2020) 146672.

[0054] [6] Y. Liu, Z. Lu, Long non-coding RNA NEAT1 mediates the toxic ofParkinson's disease induced by MPTP / MPP+ via regulation of gene expression,Clin Exp Pharmacol Physiol, 45 (2018) 841-848。

Claims

1. LncRNA NEAT1 is used as a marker for the diagnosis of Parkinson's disease, so as to further diagnose whether the structure and function of the substantia nigra region of the midbrain are normal and whether Parkinson's disease exists, and can be used to develop diagnostic agents or kits related to Parkinson's disease.

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