Application of substance targeting hsf-1 gene in preparation of medicine for treating or preventing amyotrophic lateral sclerosis

Abstract

The invention belongs to the technical field of gene functions and application, and particularly relates to application of a substance targeting an hsf-1 gene to preparation of a medicine for treating or preventing amyotrophic lateral sclerosis. According to the invention, an amyotrophic lateral sclerosis nematode model is constructed, a regulatory factor hsf-1 gene of amyotrophic lateral sclerosis is screened by adopting a transcriptome sequencing method, and the regulation effect of the hsf-1 gene on amyotrophic lateral sclerosis is verified from multiple angles. The result of the embodiment shows that the hsf-1 gene has an adjusting effect on the life, the athletic ability, the motor neuron apoptosis, the toxic protein degradation and the metabolic remodeling of the amyotrophic lateral sclerosis nematode, and further shows that the hsf-1 gene can be used for clinical rapid diagnosis of the amyotrophic lateral sclerosis and can also be used for rapid diagnosis of the amyotrophic lateral sclerosis. The development of related drugs for treating amyotrophic lateral sclerosis can be promoted, and the application has a wide development prospect and application value.

Description

Technical Field

[0001] The present invention belongs to the technical field of gene function and application, and particularly relates to the application of a substance targeting the hsf-1 gene in the preparation of a drug for treating or preventing amyotrophic lateral sclerosis. Background Art

[0002] Amyotrophic lateral sclerosis (ALS) is a sporadic neurodegenerative disease that mainly affects motor neurons, leading to muscle weakness and atrophy. Existing treatment methods are mostly symptomatic treatments, and there is no effective cure yet.

[0003] In recent years, researchers have begun to focus on Caenorhabditis elegans as a model organism and use its simple nervous system and characteristics of easy genetic manipulation to screen for ALS-related genes and regulatory factors. Existing studies have shown that Caenorhabditis elegans can effectively simulate certain characteristics of human neurodegenerative diseases, providing new ideas for the mechanism research and drug screening of ALS.

[0004] Currently, ALS research based on Caenorhabditis elegans mainly focuses on the exploration of gene functions and drug screening. Researchers construct ALS model nematodes through transgenic technology to observe the effects of specific gene mutations on motor neurons. In addition, using high-throughput screening technology, researchers can quickly evaluate the protective effects of compounds on ALS models. Although existing technologies have made certain progress in screening potential drugs, there are still problems such as low screening efficiency and poor specificity, which limit their popularization in clinical applications.

[0005] Although many studies have used Caenorhabditis elegans to screen for ALS-related factors, existing methods often lack systematicness, resulting in insufficient repeatability and reliability of screening results. In addition, existing screening methods usually only focus on a single factor and fail to comprehensively evaluate the interactions between multiple factors and their comprehensive effects on the ALS process. These problems make it difficult to translate current research results into effective clinical treatment plans.

[0006] Therefore, there is an urgent need for a new ALS regulatory factor to provide new ideas and strategies for the treatment of ALS. Summary of the Invention

[0007] The object of the present invention is to provide the application of a substance targeting the hsf-1 gene in the preparation of a drug for treating or preventing amyotrophic lateral sclerosis. The hsf-1 gene provided by the present invention not only provides a new target for the early diagnosis, treatment, and new drug research and development of amyotrophic lateral sclerosis, but also can reduce the aggregation of toxic proteins in ALS patients and protect motor neurons by stimulating the regulatory function of the hsf-1 gene, so as to improve the quality of life and extend the lifespan of patients.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] The present invention provides an application of a substance targeting the hsf-1 gene in the preparation of a medicament for treating or preventing amyotrophic lateral sclerosis.

[0010] Preferably, the expression of the hsf-1 gene is down-regulated in patients with amyotrophic lateral sclerosis.

[0011] Preferably, the medicament for treating or preventing amyotrophic lateral sclerosis up-regulates the expression of the hsf-1 gene.

[0012] Preferably, the DNA sequence of the hsf-1 gene is as shown in the nucleotide sequence SEQ ID NO.1.

[0013] The present invention also provides a pharmaceutical composition, which targets the hsf-1 gene and further comprises at least one pharmaceutically acceptable carrier, diluent and excipient.

[0014] Preferably, the pharmaceutical composition increases the expression level of the hsf-1 gene.

[0015] The present invention also provides an application of a substance targeting the hsf-1 gene in the preparation of a medicament for protecting motor neurons.

[0016] The present invention also provides an application of a substance targeting the hsf-1 gene in the preparation of a medicament for promoting the degradation of the toxic protein Q40.

[0017] The present invention also provides an application of a substance targeting the hsf-1 gene in the preparation of a medicament for regulating metabolic remodeling.

[0018] Advantages of the present invention:

[0019] The hsf-1 gene provided by the present invention provides a new target for the early diagnosis, treatment and new drug research and development of amyotrophic lateral sclerosis.

[0020] By activating the regulatory function of the hsf-1 gene of the present invention, the aggregation of toxic proteins in ALS patients can be reduced, motor neurons can be protected, so as to improve the quality of life and prolong the life of patients.

[0021] The regulatory factor hsf-1 gene of amyotrophic lateral sclerosis provided by the present invention can not only be used for the rapid diagnosis of clinical amyotrophic lateral sclerosis, but also promote the development of drugs related to the treatment of amyotrophic lateral sclerosis, and has broad development prospects and application values. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0023] Figure 1 Results graph of the screening process of regulators for amyotrophic lateral sclerosis; (A) PCA graph of the control group and ALS strain samples; (B) volcano graph; (C) heat map;

[0024] Figure 2 Results graph of the regulatory effect of regulators for amyotrophic lateral sclerosis on the lifespan and motor ability of ALS nematodes; (A-C) are lifespan curves of different nematodes; (D-G) are graphs representing the motor ability of nematodes (N2 is the wild-type control group; hsf-1(+) is the hsf-1 overexpression group; hsf-1(-) is the hsf-1 knockdown group);

[0025] Figure 3 Results graph of the regulatory effect of the hsf-1 gene on the apoptosis of motor neurons in ALS nematodes; (A) Fluorescence schematic diagram of the regulatory effect of the hsf-1 gene on lipid expression and the number of motor neurons in ALS nematodes; (B) Statistical graph of the regulatory effect of the hsf-1 gene on lipid expression in ALS nematodes; (C) Statistical graph of the regulatory effect of the hsf-1 gene on the number of motor neurons in ALS nematodes (hsod1 is the ALS control group; hsod1hsf-1(+) is the ALS hybrid hsf-1 overexpression group; hsod1hsf-1(-) is the ALS hybrid hsf-1 knockdown group);

[0026] Figure 4 Results graph of the regulatory effect of the hsf-1 gene on the degradation of toxic proteins in AM141 nematodes; (A) Fluorescence schematic diagram of the regulatory effect of the hsf-1 gene on lipid and polyglutamine (polyQ, Q40) expression in AM141 nematodes; (B) Statistical graph of the regulatory effect of the hsf-1 gene on lipid expression in AM141 nematodes; (C) Statistical graph of the regulatory effect of the hsf-1 gene on Q40 expression in AM141 nematodes; (Q40 is the AM141 control group; Q40hsf-1(+) is the AM141 hybrid hsf-1 overexpression group; Q40hsf-1(-) is the AM141 hybrid hsf-1 knockdown group);

[0027] Figure 5It is a characterization result graph of the regulatory effect of hsf-1 gene on different categories of metabolites in ALS nematodes, including fatty acyls, glycerophospholipids, sphingolipids, amino acid analogues, amino acids, purines and pyrimidines (hsod1 is the ALS control group; hsod1hsf-1(+) is the ALS hybrid hsf-1 overexpression group; hsod1hsf-1(-) is the ALS hybrid hsf-1 knockdown group). Detailed implementation mode

[0028] The present invention provides an application of a substance targeting hsf-1 gene in the preparation of a drug for treating or preventing amyotrophic lateral sclerosis. By constructing an amyotrophic lateral sclerosis model based on Caenorhabditis elegans, an amyotrophic lateral sclerosis model of Caenorhabditis elegans in the same growth cycle is prepared, and the regulatory factor hsf-1 of amyotrophic lateral sclerosis is screened by transcriptome sequencing. The expression of the regulatory factor hsf-1 is significantly down-regulated in the samples of the amyotrophic lateral sclerosis strain.

[0029] Overexpression can be achieved by conventional methods in the art, including plasmid vector-mediated, constructing a viral vector system, and chromosomal integration technology, etc. Preferably, it is to construct a viral vector system.

[0030] The substance targeting hsf-1 gene of the present invention is applicable to all humans or animals containing hsf-1 gene, and the present invention is applied to the amyotrophic lateral sclerosis model of Caenorhabditis elegans.

[0031] The screening method of the regulatory factor of amyotrophic lateral sclerosis based on Caenorhabditis elegans of the present invention specifically includes: constructing an amyotrophic lateral sclerosis model based on Caenorhabditis elegans, designing and constructing a human-derived SOD1-G93A plasmid with GFP tag under the control of unc-25 promoter based on the N2 nematode genome; constructing an amyotrophic lateral sclerosis model of Caenorhabditis elegans by the method of microinjection combined with ultraviolet irradiation-assisted genome fusion, wherein the plasmid concentration is 100 ng / μL; backcrossing with N2 nematodes 6 times to obtain a Caenorhabditis elegans strain stably expressing human-derived SOD1(G93A) protein. By culturing Escherichia coli OP50; coating Escherichia coli OP50 on NGM culture plates; culturing Caenorhabditis elegans on NGM plates; synchronizing Caenorhabditis elegans by sodium hypochlorite lysis method, and finally obtaining an amyotrophic lateral sclerosis model of Caenorhabditis elegans in the same growth cycle.

[0032] The UNC-25 promoter is a key tool for achieving specific expression in GABAergic neurons in the study of Caenorhabditis elegans. This promoter can precisely drive the expression of foreign genes in GABAergic neurons such as D-type motor neurons. These neurons inhibit muscle contraction by releasing γ-aminobutyric acid (GABA) and play an important role in coordinating the movement patterns of C. elegans. In neurobiological research, this promoter is often used to construct transgenic C. elegans models for conditional gene expression or silencing in specific neuronal subsets.

[0033]

[0034] Based on Caenorhabditis elegans with amyotrophic lateral sclerosis, nematode models with overexpression and knockdown of hsf-1 were constructed respectively, and the regulatory effect of the hsf-1 gene on amyotrophic lateral sclerosis was verified from multiple perspectives. The results of the examples show that the hsf-1 gene has a regulatory effect on the lifespan, motor ability, motor neuron apoptosis, toxic protein degradation, and metabolic remodeling of Caenorhabditis elegans with amyotrophic lateral sclerosis. Overexpression of the hsf-1 gene can significantly extend the lifespan of Caenorhabditis elegans; overexpression of the hsf-1 gene has a significant protective effect on the motor ability of Caenorhabditis elegans; overexpression of the hsf-1 gene can significantly increase the number of motor neurons in Caenorhabditis elegans; overexpression of the hsf-1 gene has an obvious inhibitory effect on the aggregation of toxic proteins in Caenorhabditis elegans; overexpression of the hsf-1 gene shows a significant accumulation of fatty acyls, glycerophospholipids, sphingolipids, and amino acid analogues, as well as a decrease in amino acids, purines, and pyrimidines in Caenorhabditis elegans.

[0035] Furthermore, it is shown that the amyotrophic lateral sclerosis regulatory factor hsf-1 provided by the present invention can not only be used for the rapid diagnosis of clinical amyotrophic lateral sclerosis, but also promote the development of drugs related to the treatment of amyotrophic lateral sclerosis, and has broad development prospects and application values.

[0036] The GFP-tag is a technique that uses green fluorescent protein (GFP) as a tag for labeling and visualizing target proteins.

[0037] Oligo(dT) magnetic beads are 1μm magnetic polymer microspheres coated with Oligo(dT) on the surface, with characteristics such as monodispersity and strong magnetic responsiveness. They can quickly and efficiently isolate and purify mRNA from total RNA, cell lysates, and in vitro transcribed RNA samples with polyA tails through the complementary pairing of Oligo(dT) with the polyA tail structure of eukaryotic mRNA.

[0038] The SOD1-G93A protein is a mutant human superoxide dismutase 1 (SOD1) protein, in which glycine at position 93 is mutated to alanine (G93A). This mutation leads to abnormal function of the SOD1 protein, thereby triggering a series of pathological changes, and is mainly used for studying the pathogenesis of amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig's disease).

[0039] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below in conjunction with the drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0040] The mirVana TM The miRNA ISOlation Kit was purchased from the Ambion-1561 brand; the nematodes in the hsf-1 overexpression group and the hsf-1 knockdown group are commercially available strains.

[0041] In the embodiments of the present invention, the production processes, experimental methods or detection methods involved, unless otherwise specified, are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the art and are very clear and definite in the relevant application fields. Those skilled in the art can understand the conventional process steps according to the name and apply the corresponding equipment and implement them under conventional conditions or conditions recommended by the manufacturer.

[0042] There are no special restrictions on the sources of various instruments, equipment, raw materials or reagents used in the embodiments of the present invention. They are all conventional products that can be obtained through regular commercial channels, and can also be prepared according to the conventional methods well-known to those skilled in the art.

[0043] Example 1 Construction of an amyotrophic lateral sclerosis model based on Caenorhabditis elegans

[0044] Based on the N2 nematode genome, a human SOD1-G93A plasmid with GFP tag under the control of the unc-25 promoter was designed and constructed; an amyotrophic lateral sclerosis model of Caenorhabditis elegans was constructed by microinjection combined with ultraviolet irradiation-assisted genome integration.

[0045] Among them, the plasmid concentration was 100 ng / μL; after backcrossing with N2 nematodes 6 times, a Caenorhabditis elegans strain stably expressing human SOD1-G93A protein was obtained.

[0046] The preparation of a Caenorhabditis elegans model in the same growth cycle includes: culturing Escherichia coli OP50; coating Escherichia coli OP50 onto an NGM culture plate; culturing Caenorhabditis elegans on the NGM plate; synchronizing Caenorhabditis elegans by the sodium hypochlorite lysis method, and finally obtaining a Caenorhabditis elegans amyotrophic lateral sclerosis model in the same growth cycle.

[0047] Example 2 Total RNA Extraction and Transcriptome Sequencing Analysis of Caenorhabditis elegans

[0048] 2.1 Total RNA Extraction, Purification and Library Construction of Caenorhabditis elegans

[0049] Approximately 10,000 Caenorhabditis elegans strains in the control group that cannot stably express human SOD1-G93A protein and Caenorhabditis elegans strains (ALS strains) that stably express human SOD1-G93A protein (volume approximately 0.3 cm × 0.3 cm × 0.3 cm) were collected respectively and stored at -80 °C for later use.

[0050] According to the steps of the mirVana TM miRNA ISOlation Kit, Ambion-1561 kit, total RNA of nematode samples was extracted and DNA was digested with DNase. Then, eukaryotic mRNA was enriched with magnetic beads with Oligo(dT). The mRNA was fragmented by adding fragmentation reagent. Using the fragmented mRNA as a template, first-strand cDNA was synthesized with hexamer random primers, and then a second-strand synthesis reaction system was prepared to synthesize second-strand cDNA, and the double-stranded cDNA was purified using the kit. The purified double-stranded cDNA was then subjected to end repair, A-tailing and ligation of sequencing adapters, followed by fragment size selection, and finally PCR amplification. After the constructed library passed the quality inspection with an Agilent 2100 Bioanalyzer, Illumina HiSeqTM2500 was used for sequencing to generate 125bp or 150bp paired-end data.

[0051] 2.2 Bioinformatics Analysis of Sequencing Data

[0052] The raw data obtained by sequencing was subjected to quality filtering and cleaning, and the estimateSizeFactors function of the R package was used to standardize the data. The pvalue and FoldChange values for differential comparison were calculated, and genes with a p-value less than 0.05 and a differential multiple greater than 2 were selected for subsequent analysis.

[0053] The results are as shown in Figure 1 (A). The visualization result PCA plot of principal component analysis shows that the samples of the control group and ALS strains have a good separation effect, proving that the separation method applied in this project is feasible.

[0054] Visualize the differential genes using a volcano plot, and the results are as Figure 1 (B) shown. Compared with the control group, there are 111 genes with up-regulated expression in the ALS strain nematode samples, including the gmd-2, fmo-2, and fat-7 genes; there are 64 genes with down-regulated expression, including the hsf-1, jnk-1, sod-5, alh-2, hsp-12.6, ant-1.2, and smf-3 genes, etc.

[0055] Subsequently, perform unsupervised hierarchical clustering on the significantly differential genes (p < 0.05, log2FC > 2), and use the form of a heat map to display the expression patterns of the differential genes among different samples. The results are as Figure 1 (C) shown. It is found that among all the differential genes, the hsf-1 gene changes most significantly (log2FC = -5.13), indicating that the hsf-1 gene may play an obvious regulatory role in the disease process of amyotrophic lateral sclerosis.

[0056] Example 3 Functional evaluation of the regulatory role of the hsf-1 gene in amyotrophic lateral sclerosis

[0057] Using the nematode hybridization technology mastered in the laboratory, transgenic nematode hybrid strains with different functional protein expressions are constructed respectively, and then corresponding functional characterizations are carried out to verify the regulatory role of the hsf-1 gene in the disease process and development trend of amyotrophic lateral sclerosis.

[0058] 3.1 Characterization of the regulatory role of the hsf-1 gene in the lifespan and behavior of nematodes with amyotrophic lateral sclerosis

[0059] Set wild-type control group, hsf-1 overexpression group, and hsf-1 knockdown group nematodes, and investigate the lifespan of the three groups of nematodes respectively. The hsf-1 overexpression group and hsf-1 knockdown group nematodes are commercially available strains.

[0060] The results are as Figure 2 (A-C) shown. It is found that the lifespan of the hsf-1 overexpression group nematodes is significantly prolonged, and the lifespan of the hsf-1 knockdown group nematodes is significantly shortened, proving that hsf-1 has the function of prolonging the lifespan of Caenorhabditis elegans. Subsequently, set ALS control group, ALS hybrid hsf-1 overexpression group, and ALS hybrid hsf-1 knockdown group, and investigate the lifespan of the three groups of nematodes respectively. It is also found that hsf-1 overexpression plays a significant role in prolonging the lifespan of ALS nematodes, and the lifespan of the hsf-1 knockdown group ALS nematodes is significantly shortened, proving that hsf-1 also has the function of prolonging the lifespan of Caenorhabditis elegans with amyotrophic lateral sclerosis.

[0061] Characterize the motor behavior of the ALS group nematodes. The results are as Figure 2(As shown in (D-G), it was found that the motor ability of ALS nematodes in the hsf-1 overexpression group was significantly restored, while that of ALS nematodes in the hsf-1 knockdown group did not improve significantly, indicating that the expression of the hsf-1 gene has a significant protective effect on the motor ability of ALS nematodes.)

[0062] 3.2 Protective effect of the hsf-1 gene on motor neurons in amyotrophic lateral sclerosis nematodes

[0063] First, hybrid nematodes were constructed. L4-stage male ALS nematodes were obtained by heat shock at 30 °C. At the same time, hsf-1(+) or hsf-1(-) hermaphrodite nematodes of the same growth stage were selected and co-cultured with nematodes at a ratio of 1:3 (ALS: hsf-1) in an environment at 20 °C. After 24 hours, the hermaphrodite nematodes were picked out and cultured separately. After 72 hours, the nematodes with GFP labeling in the first-generation nematodes were picked out and cultured separately. After 72 hours, the nematodes with GFP fluorescence labeling in the second-generation nematodes were picked out and cultured separately. After 72 hours, nematodes without GFP labeling in the offspring were excluded, and the remaining third-generation nematodes were picked out and cultured separately. After amplification, nematodes without GFP fluorescence labeling in the offspring were excluded, and the fourth-generation nematodes with all offspring carrying GFP were selected for genotype identification. Nematodes carrying both hSOD(G93A) and hsf-1(+) or hsf-1(-) genes were the required hybrid strains.)

[0064] The number of motor neurons in the bodies of nematodes in the ALS control group, ALS hybrid hsf-1 overexpression group, and ALS hybrid hsf-1 knockdown group was examined. The results are as Figure 3 (shown, and it was found that the number of motor neurons in the bodies of ALS nematodes in the hsf-1 overexpression group increased significantly and apoptosis decreased, while the number of motor neurons in the bodies of ALS nematodes in the hsf-1 knockdown group did not change significantly compared with the control group, indicating that the expression of the hsf-1 gene has a significant inhibitory effect on the apoptosis of motor neurons in ALS nematodes, and thus plays a role in repairing motor ability.)

[0065] 3.3 Regulatory effect of the hsf-1 gene on the toxic protein Q40 in AM141 nematodes

[0066] First, hybrid nematodes were constructed. Male AM141 nematodes at the L4 stage were obtained by heat shock at 30°C. At the same time, hermaphrodite nematodes of hsf-1(+) or hsf-1(-) at the same growth stage were selected. The nematodes were co-cultured in an environment at 20°C according to the ratio of 1:3 (AM141: hsf-1). After 24 hours, the hermaphrodite nematodes were picked out and cultured separately. After 72 hours, the nematodes with GFP markers in the first-generation nematodes were picked out and cultured separately. After 72 hours, the nematodes with GFP fluorescence markers in the second-generation nematodes were picked out and cultured separately. After 72 hours, nematodes without GFP markers in the offspring were excluded, and the remaining third-generation nematodes were picked out and cultured separately. After amplification, nematodes without GFP fluorescence markers in the offspring were excluded, and the fourth-generation nematodes with all offspring carrying GFP were selected for genotype identification. Those with both Q40 and hsf-1(+) or hsf-1(-) genes were the required hybrid worm strains.

[0067] The expression levels and aggregation of polyglutamine (Q40) in the nematodes of the AM141 control group, the AM141 hybrid hsf-1 overexpression group, and the AM141 hybrid hsf-1 knockdown group were examined. The results are as Figure 4 shown. It was found that the content and the number of aggregation points of Q40 in the AM141 nematodes of the hsf-1 overexpression group were significantly reduced, while the content of Q40 and the number of aggregation points in the AM141 nematodes of the hsf-1 knockdown group did not change significantly compared with the control group. This indicates that the expression of the hsf-1 gene has an obvious inhibitory effect on the aggregation of toxic proteins in AM141 nematodes, and thus plays a role in delaying the disease process of amyotrophic lateral sclerosis.

[0068] Example 4 Analysis of the regulatory mechanism of the hsf-1 gene on amyotrophic lateral sclerosis based on metabolomics

[0069] Using the method of liquid chromatography-mass spectrometry, the relative contents of non-target metabolites in the nematodes of the ALS control group, the ALS hybrid hsf-1 overexpression group, and the ALS hybrid hsf-1 knockdown group were detected respectively.

[0070] The results are as Figure 5 shown. It was found that there were significant differences in 6 types of metabolites between the ALS control group and the ALS hybrid hsf-1 overexpression group. Compared with ALS nematodes, the ALS hybrid hsf-1 overexpression group of nematodes showed significant accumulation of fatty acyls, glycerophospholipids, sphingolipids, and amino acid analogues, as well as a decrease in amino acids, purines, and pyrimidines.

[0071] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Use of a substance targeting the hsf-1 gene in the preparation of a drug for treating or preventing amyotrophic lateral sclerosis.

2. The application according to claim 1, characterized in that: The expression of hsf-1 gene is downregulated in the ALS patients.

3. The application according to claim 1, characterized in that: The drug for treating or preventing amyotrophic lateral sclerosis upregulates the expression of the hsf-1 gene.

4. The use according to claim 1, characterized in that: The DNA sequence of the hsf-1 gene is shown in the nucleotide sequence SEQ ID NO.

1.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition takes the hsf-1 gene as a target and further comprises at least one pharmaceutically acceptable carrier, diluent and excipient.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition increases the expression level of the hsf-1 gene.

7. Application of substances targeting hsf-1 gene in the preparation of drugs for protecting motor neurons.

8. Application of substances targeting hsf-1 gene in the preparation of drugs that promote the degradation of toxic protein Q40.

9. Application of substances targeting hsf-1 gene in the preparation of drugs for regulating metabolic reconstruction.

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