CN116650652A - Diagnosis and treatment marker for diabetic peripheral neuropathy and application thereof - Google Patents

Diagnosis and treatment marker for diabetic peripheral neuropathy and application thereof Download PDF

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CN116650652A
CN116650652A CN202310886788.XA CN202310886788A CN116650652A CN 116650652 A CN116650652 A CN 116650652A CN 202310886788 A CN202310886788 A CN 202310886788A CN 116650652 A CN116650652 A CN 116650652A
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eif6
peripheral neuropathy
diabetic peripheral
expression
treatment
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焦洋
殷玲
房崇亮
李依泽
于泳浩
谢克亮
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Tianjin Medical University General Hospital
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Abstract

The invention discloses a diagnosis and treatment marker of diabetic peripheral neuropathy and application thereof, wherein the diagnosis and treatment marker is eIF6. According to the invention, the eIF6 up-regulation in the Schwann cells is closely related to diabetic peripheral neuropathy, and targeted inhibition by taking the eIF6 as a target point can well inhibit the development of the neuropathy, so that the method has important significance for early diagnosis and treatment of DPN and disease course reversal. The invention develops a novel method for diagnosing diabetic peripheral neuropathy and a novel drug target for treating the disease.

Description

Diagnosis and treatment marker for diabetic peripheral neuropathy and application thereof
Technical Field
The invention belongs to the field of clinical diagnosis and treatment, and relates to a diagnosis and treatment marker for diabetic peripheral neuropathy and application thereof.
Background
Diabetic peripheral neuropathy (Diabetic Peripheral Neuropathy, DPN) is the most common complication of diabetes, with up to 50% of diabetics, the leading cause of high disability, cardiovascular risk and mortality in diabetes. DPN is mainly represented by intractable pain, numbness and sensory disturbance distributed symmetrically at the far end of limbs, long-term illness can cause mental and psychological problems such as depression, anxiety, sleep disturbance and the like, and late stage even faces foot ulcers and lower limb amputation, so that the life quality of patients is seriously influenced. About 670 ten thousand adults worldwide die from diabetes mellitus and its complications in 2021, accounting for 12.2% of the total worldwide death population, corresponding to about 1 death every 5 seconds, and one person facing amputation due to diabetic foot every 30 seconds. However, as the pathogenic mechanism is not completely elucidated, DPN is not yet provided with special drugs approved by FDA, clinical treatment is limited to controlling sugar and easing pain mainly, and the disease process can not be effectively delayed, so that the incidence of DPN is still rapidly increased in a situation of being difficult to control, and huge economic and social burdens are brought. Therefore, the exact pathogenesis of DPN is clarified as early as possible, and the development of drugs capable of effectively delaying the development of the course of disease is unprecedented.
Disclosure of Invention
The invention discloses the use of eIF6 gene in preparing medicine composition for preventing and treating diabetic peripheral neuropathy.
Further, the prevention or treatment of diabetic peripheral neuropathy refers to prevention or treatment of hypoalgesia in diabetic peripheral neuropathy.
Preferably, the pain sensation comprises mechanical pain sensation and thermal pain sensation.
Further, the pharmaceutical composition comprises an inhibitor of eIF6 functional expression.
Further, the inhibitor is an agent that knocks out eIF6 by CRISPR/Cas9 technology.
In a second aspect, the invention discloses a pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, the pharmaceutical composition comprising an inhibitor of eIF6 functional expression.
Such inhibitors include, but are not limited to, antagonists, blockers, nucleic acid inhibitors, and the like.
Inhibitors refer to any substance that decreases the activity of the eIF6 protein, decreases the stability of the eIF6 gene or protein, down regulates the expression of the eIF6 protein, decreases the effective duration of the eIF6 protein, or inhibits the transcription and translation of the eIF6 gene, which can be used in the present invention as a substance useful for down regulating eIF6, and thus can be used for preventing or treating diabetic peripheral neuropathy.
By inhibitors of the eIF6 gene or protein is meant any agent that reduces the activity of the eIF6 protein, reduces the stability of the eIF6 gene or protein, down regulates the expression of the eIF6 protein, reduces the effective duration of the eIF6 protein, or inhibits the transcription and translation of the eIF6 gene, which is useful as an agent for down regulating eIF6 and thus can be used in the prevention or treatment of diabetic peripheral neuropathy. For example, the inhibitors are: nucleic acid inhibitors, protein inhibitors, antibodies, ligands, proteolytic enzymes, protein binding molecules, provided that they are capable of down-regulating expression of eIF6 proteins or genes encoding them at the protein or gene level.
As a preferred mode of the invention, the inhibitor of eIF6 is an eIF6 specific small interfering RNA molecule. As used herein, the term "small interfering RNA" refers to a short segment of double-stranded RNA molecule capable of degrading a specific mRNA targeting the mRNA of homologous complementary sequence, which is the RNA interference (RNA interference) process. The small interfering RNA can be prepared in the form of a double-stranded nucleic acid comprising a sense strand and an antisense strand, which form a double strand only under hybridization conditions. A double stranded RNA complex can be prepared from the sense strand and the antisense strand separated from each other. Thus, for example, the complementary sense and antisense strands are chemically synthesized, and can be subsequently hybridized by annealing to produce a synthetic double stranded RNA complex.
As an alternative to the present invention, the eIF6 inhibitor may also be a "Small hairpin RNA (shRNA)", which is a non-coding Small RNA molecule capable of forming a hairpin structure, which is capable of inhibiting gene expression via the RNA interference pathway. As described above, shRNA may be expressed from a double stranded DNA template. The double stranded DNA template is inserted into a vector, such as a plasmid or viral vector, and then ligated to a promoter for expression in vitro or in vivo. shRNA can be cleaved into small interfering RNA molecules by the action of DICER enzyme in eukaryotic cells, thereby entering the RNAi pathway. "shRNA expression vector" refers to a number of plasmids conventionally used in the art to construct shRNA structures, typically having a "spacer" and multiple cloning sites or alternative sequences flanking the "spacer" such that one can insert the corresponding DNA sequence of the shRNA (or analog) into the multiple cloning site or alternative sequences thereon in a forward and reverse manner, and the RNA transcribed from the DNA sequence can form a shRNA (Short Hairpin) structure. The "shRNA expression vectors" are now fully commercially available, for example, as some viral vectors.
The nucleic acid inhibitors of the invention, such as siRNA, may be chemically synthesized or prepared by transcription of an expression cassette in a recombinant nucleic acid structure into single stranded RNA. Nucleic acid inhibitors such as siRNA can be delivered into cells by use of an appropriate transfection reagent, or can also be delivered into cells using a variety of techniques known in the art.
In a specific embodiment of the invention, the inhibitor is an agent that knocks out eIF6 by CRISPR/Cas9 technology.
Further, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
The term "pharmaceutically acceptable carrier" refers to a carrier for administration of a therapeutic agent, including various excipients and diluents. The term refers to such agent carriers: they are not per se essential active ingredients and are not overly toxic after administration. Suitable vectors are well known to those of ordinary skill in the art. The pharmaceutically acceptable carrier in the composition may contain a liquid, such as water, saline, buffer. In addition, auxiliary substances such as fillers, lubricants, glidants, wetting or emulsifying agents, pH buffering substances and the like may also be present in these carriers. The vector may also contain a cell (host cell) transfection reagent.
The present invention may employ a variety of methods well known in the art for administering the inhibitor or gene encoding the same, or a pharmaceutical composition thereof, to a mammal. Including but not limited to: subcutaneous injection, intramuscular injection, transdermal administration, topical administration, implantation, sustained release administration, and the like; preferably, the mode of administration is parenteral.
Preferably, gene therapy is used. For example, an inhibitor of eIF6 may be administered directly to a subject by a method such as injection; alternatively, expression units (e.g., expression vectors or viruses, or siRNA or shRNA) carrying an inhibitor of eIF6 may be delivered to the target site by a route that allows for expression of the active eIF6 inhibitor, depending on the type of inhibitor, as is well known to those skilled in the art.
The pharmaceutical compositions of the invention may also be used in combination with other drugs for the treatment of diabetic peripheral neuropathy, and other therapeutic compounds may be administered simultaneously with the primary active ingredient, even in the same composition.
The pharmaceutical compositions of the present invention may also be administered alone with other therapeutic compounds, either in a separate composition or in a dosage form different from the primary active ingredient. A partial dose of the principal component may be administered simultaneously with other therapeutic compounds, while other doses may be administered separately. The dosage of the pharmaceutical composition of the present invention may be adjusted during the course of treatment according to the severity of the symptoms, the frequency of recurrence and the physiological response of the treatment regimen.
In a third aspect, the invention discloses the use of the eIF6 gene for screening candidate compounds for the prevention or treatment of diabetic peripheral neuropathy.
Further, the step of screening candidate compounds for preventing or treating diabetic peripheral neuropathy comprises:
in a test group, adding a test compound into a culture system of cells, and observing the expression amount and/or activity of eIF6 in the cells of the test group; in the control group, no test compound is added in the culture system of the same cells, and the expression amount and/or activity of eIF6 in the cells of the control group are observed; wherein, if the expression level and/or activity of eIF6 in the cells in the test group is lower than that in the control group, the test compound is a candidate compound for treating diabetic peripheral neuropathy having an inhibitory effect on expression and/or activity of eIF6.
In the present invention, the steps further include: further cell and/or animal experiments are performed on the candidate compounds obtained to further select and identify substances from the candidate compounds that are useful for preventing, alleviating or treating diabetic peripheral neuropathy.
In the present invention, the system for screening candidate compounds for preventing or treating diabetic peripheral neuropathy is not limited to a cell system, but includes a cell system, a subcellular system, a solution system, a tissue system, an organ system, an animal system, or the like, and is not limited to the above-described form as long as the system can detect a test compound to reduce the expression and/or activity of eIF6.
In a fourth aspect, the invention discloses the use of an agent for detecting the expression level of eIF6 genes in the preparation of a product for diagnosing diabetic peripheral neuropathy.
The present invention can determine gene expression using any method known in the art. It will be appreciated by those skilled in the art that the means for determining gene expression is not an important aspect of the present invention. Exemplary methods of quantifying RNA expression in a sample known in the art include, but are not limited to, southern blotting, northern blotting, microarray, polymerase Chain Reaction (PCR), NASBA, and TMA.
Further, the agent is selected from: a probe that specifically recognizes eIF 6; or a primer that specifically amplifies eIF 6; or an antibody or ligand that specifically binds to a protein encoded by eIF6.
Further, the product includes a chip or a kit.
The chip may be a gene chip, and the gene chip of the present invention includes: a solid phase carrier; and an oligonucleotide probe orderly immobilized on the solid support, wherein the oligonucleotide probe specifically corresponds to a part or all of the sequence shown in eIF6.
Specifically, suitable probes can be designed according to the genes of the invention and immobilized on a solid support to form an "oligonucleotide array". By "oligonucleotide array" is meant an array having addressable locations (i.e., locations characterized by distinct, accessible addresses), each addressable location containing a characteristic oligonucleotide attached thereto. The oligonucleotide array may be divided into a plurality of subarrays, as desired.
Probes "are intended to include nucleic acid oligomers or aptamers that specifically hybridize to a target sequence in a nucleic acid or its complement under conditions that promote hybridization, thereby allowing detection of the target sequence or its amplified nucleic acid. Detection may be direct (i.e., generated by a probe that hybridizes directly to the target or amplified sequence) or indirect (i.e., generated by a probe that hybridizes to an intermediate molecular structure linking the probe and the target or amplified sequence). The "target" of a probe generally refers to a sequence in an amplified nucleic acid sequence that specifically hybridizes to at least a portion of the probe sequence via standard hydrogen bonding or "base pairing". The "sufficiently complementary" sequence allows stable hybridization of the probe sequence to the target sequence even if the two sequences are not fully complementary. The probe may be labeled or unlabeled. Probes can be produced by molecular cloning of specific DNA sequences, or can be synthesized. Those skilled in the art to which the present invention pertains can readily determine a variety of primers and probes that can be designed and used in the context of the present invention.
The solid phase carrier of the present invention may be made of various materials commonly used in the field of gene chips, including, but not limited to, plastic products, microparticles, membrane carriers, etc. The plastic product can be combined with an antibody or a protein antigen through a non-covalent or physical adsorption mechanism, and the most common plastic products are small test tubes, small beads and micro-reaction plates made of polystyrene; the microparticles are microspheres or particles polymerized by high molecular monomers, have the diameter of micrometer, are easy to form chemical coupling with antibodies (antigens) due to the functional groups capable of being combined with proteins, and have large combining capacity; the membrane carrier comprises microporous filter membranes such as nitrocellulose membranes, glass cellulose membranes and nylon membranes.
The invention has the advantages and beneficial effects that:
1) According to the invention, the eIF6 up-regulation in the Schwann cells is closely related to diabetic peripheral neuropathy, and targeted inhibition by taking the eIF6 as a target point can well inhibit the development of the neuropathy, so that the method has important significance for early diagnosis and treatment of DPN and disease course reversal.
2) The invention provides a method of preventing or treating diabetic peripheral neuropathy comprising administering to a subject an effective amount of an agent that inhibits eIF6.
Drawings
Figure 1 shows a graph of the results of expression of animal level eIF6 in DPN, wherein a: immunoblotting; b: a histogram; c: RT-qPCR diagram; n=8; * P <0.001 compared to db/m group;
fig. 2 shows a graph of the results of expression of cell level eIF6 in DPN, wherein a: immunoblotting; b: a histogram; c: RT-qPCR diagram; * P <0.001 compared to Control group;
figure 3 shows a graph of the results of eIF6 effect on DPN treatment, wherein a: mechanical foot-reduction threshold (PWT); b: thermal foot latency (TWL); n=8; * p <0.05, < p <0.01, < p <0.001, compared to db/m group; # p <0.01, # p <0.001, compared to db/db group; two-way ANOVA.
Detailed Description
The invention will now be described in further detail with reference to the drawings and examples. The following examples are only illustrative of the present invention and are not intended to limit the scope of the invention. The experimental procedure, without specific conditions noted in the examples, is generally followed by conventional conditions, such as Sambrook et al, molecular cloning: conditions described in the laboratory Manual (New York: cold Spring HarborLaboratoryPress, 1989) or as recommended by the manufacturer.
Example eIF6 expression correlation with DPN
1. Experimental method
(1) Animal experiment group: the healthy male mice of C57 BSK db/db (classical model of type II diabetes) of 16 weeks old are used as animal models of diabetic peripheral neuropathy, db/m mice of the same nest are used as normal control groups, 16 mice are provided by Hengzhou Karns laboratory animal Co., ltd., and after the model is confirmed by fasting blood glucose and glucose tolerance test detection, the mice are divided into 4 groups (n=8) by adopting a random digital meter method:
(1) control plasmid +db/m group (db/m): a needle was inserted between the tibialis anterior and gastrocnemius, and a CRISPR/Cas9 control plasmid (SANTA CRUZ, sc-418922, usa) was injected with a microinjector for a sciatic nerve side injection of 1 μl each, and after 2 weeks of feeding, further experiments were performed.
(2) Control plasmid +db/db group (db/db): a further experiment was performed using a microinjector to inject CRISPR/Cas9 control plasmid (SANTA CRUZ, sc-418922, USA) at the sciatic parasympathetic side, 1 μl each, after 2 weeks of feeding.
(3) eIF6 ko+db/m group (e+db/m): a microinjector was used to inject eIF6CRISPR/Cas9 KO plasmid (SANTACRUZ, sc-421176, usa) at the sciatic nerve side, 1 μl each for knocking down eIF6 expression in sciatic nerve, and further experiments were performed after 2 weeks of feeding.
(4) eIF6 ko+db/db group (e+db/db): additional experiments were performed 2 weeks after injection of the eIF6CRISPR/Cas9 KO plasmid (SANTA CRUZ, sc-421176, usa) using a microinjector at the sciatic nerve side, each with 1 μl of the sciatic nerve side for knocking down eIF6 expression in the sciatic nerve.
(2) Behavioural experiments
Continuous observation was performed for 6 weeks from 16 weeks of age, and mechanical shrinkage threshold (PWT) and thermal shrinkage latency (TWL) were measured at 16, 18, 20, 22, and 24 weeks, respectively, mice were placed in a 20cm x 20cm metal cage in a quiet environment at room temperature of 22-25 ℃ to accommodate the environment for 1 hour or more, and mechanical shrinkage threshold (paw withdrawal threshold) was measured using calibrated von Frey filaments (Stoelting, usa), PWT), ranging from 0.02 to 1.4g, applying von Frey filament pressure perpendicular to the hind paw from small to large and slightly bending it for 3s each time according to the modified Up-down method described by Chaplan et al, recording the response to each stimulus, and if the mice developed tremble, shrink paw, lick the paw, then they were considered positive, measuring 6 times in succession, 15min apart, calculating the 50% mechanical shrink threshold according to the following formula record: 50% PWT=10 (xf+kδ)/10000.
The heat shrinkage foot latency (Thermal withdrawal latency, TWL) was measured using an infrared plantar thermal pain measuring instrument (IITC Life Science, woodland Hills, U.S.), the mice were placed in a transparent closed apparatus and acclimatized for at least 20 minutes, the foot shrinkage latency due to radiant heat (15% heating intensity) was recorded, at least 15min between the two measurements, 3 consecutive measurements were made, and the upper cutoff time was set at 30s to prevent scalding.
(3) Ischial nerve specimen collection
After the last behavioural measurement is finished, the mice at 24 weeks of age are sacrificed after anesthesia by using the inhalant anesthetic sevoflurane, the mice are fixed in prone position, the left lower limb is prepared for skin preparation, iodophor is disinfected, the skin is cut, the visual field is exposed, the muscles are blunt-separated, the sciatic nerve from the sciatic tuberosities to the inner side of the ankle is exposed, the sciatic nerve is dissociated and cut off, and the mice are quickly placed in liquid nitrogen for freezing and stored in a refrigerator at the temperature of minus 80 ℃.
(4) Cell culture
Rat schwann cell line (Rat Schwann cells, RSC 96) was purchased from Shanghai Tongpi derivative technologies, inc., and incubated at 37℃under constant temperature with 5% CO 2 In the incubator of (2), 10% FBS was added to DMEM medium, the medium was changed 1 time every 1 day, and the growth of schwann cells was observed under an inverted microscope, and exponentially growing cells were taken for subsequent experiments. Wherein the high sugar group cells (HG group) were cultured with 25mM glucose medium for 48 hours for simulating high sugar damage, the Control group (Control group) was cultured with low sugar medium containing 5.5mmol/L glucose for 48 hours, and then the cells were collected for subsequent experiments.
(5) Real-time PCR (QPCR) reaction
The frozen sciatic nerve tissue or schwann cells are taken out and resuscitated, total mRNA is extracted by the operation steps provided by the specification of a TRIzol kit (Invitrogen), cDNA is synthesized by referring to the specific reverse primer of the specification of the reverse transcription kit, and PCR detection is carried out by using the cDNA as a template and an ABI 7500 real-time fluorescence quantitative polymerase chain reaction instrument. According to the experimental grouping and the number of the detected genes, designing a QPCR sample adding sequence, setting 3 compound holes for each experimental sample, configuring the required reagent for each hole according to the specification, and replacing the DNA template of the negative control hole by DEPC water. And (5) after uniformly mixing, performing instantaneous centrifugation, and centering the mixture in a QPCR instrument. The reaction procedure: pre-denaturation at 95℃for 10min, 15s at 95℃for 120s at 60℃for 20s at 72℃for 30 cycles. The primer sequences of the target genes are as follows:
eIF6-F：GAGTTGTCCTCCCTTCTTCAGG(SEQ ID NO.1)；
eFI6-R：TCGTGTCCAGACCACAGAAAGC(SEQ ID NO.2)
(6)Western Blot
protein expression was determined by Western Blot. Taking out frozen ischial nerve tissue or schwann cells, adding precooled tissue protein lysate, and grinding. Centrifuging the homogenate at 4 ℃ for 15min at 13000rpm, taking supernatant, centrifuging again, repeating for 3 times again, and obtaining supernatant which is the total protein sample of the schwann cells. After preliminary quantification of the protein concentration of the samples by BCA method, expression of eIF6 was determined experimentally using rabbit-derived eIF6 antibody (HUABIO, ER2001-54, usa) and rabbit-derived GAPDH antibody (Abcam corporation, ab-181602, usa) according to the guidelines specified in the specification.
(7) Statistical analysis: data were analyzed using SPSS21.0 software or Graphpad Prism 6.0, with data normally distributed as mean.+ -. Standard deviationEach statistic is represented as an average of a minimum of three replicates. Two independent sample t-test is adopted between the two groups of data; the p value is obtained by adopting single-factor variance analysis among the multiple groups of data, and the two-by-two comparison is carried out by adopting a Turkey method; the data obtained by repeated measurement in multiple time periods adopts variance analysis of repeated measurement data, and is combined with Bonferroni method to carry out inter-group comparison after measurement, p<The difference was considered statistically significant at 0.05.
2. Experimental results
(1) mRNA and protein expression of eIF6 in sciatic nerve of db/db mice is increased
48 hours after the end of the last behavioral determination, the sciatic nerve is sacrificed from the rat, the Western Blot experiment result shows that compared with db/m group, the expression of eIF6 protein in the sciatic nerve of the db/db group of mice is obviously increased (p < 0.001), and the RT-qPCR result also shows that the mRNA level of eIF6 in the sciatic nerve of the db/db group of mice is obviously increased (p < 0.001) (the result is shown in figure 1). The above results demonstrate from the in vivo level that the occurrence of diabetic peripheral neuropathy was associated with upregulated expression of eIF6.
(2) mRNA and protein expression of eIF6 in high sugar Schwann cells
After RSC96 cells were cultured in low-sugar (Control group) or high-sugar (HG group) medium for 48 hours, western Blot experiment results showed that expression of eIF6 protein was significantly increased (p < 0.001) in HG group cells compared to Control group cells, and RT-qPCR results also showed that mRNA level of eIF6 was significantly increased (p < 0.001) in HG group cells (see fig. 2 for results). The above results demonstrate from an ex vivo level that the occurrence of diabetic peripheral neuropathy was associated with upregulated expression of eIF6.
(3) eIF6 knockout inhibited mechanical allodynia and thermal hypodynia in db/db mice
The results are shown in FIG. 3, in which the mechanical foot-shrinking threshold PWT (g) is significantly increased (p < 0.001) in db/db groups compared with db/m groups, the thermal foot-shrinking latency TWL(s) is significantly prolonged, and hypoalgesia occurs, indicating that the diabetic peripheral neuropathy model is established, within 16-24 weeks. No significant changes (p > 0.05) were seen in PWT and TWL in the E+db/m group compared to the db/m group, indicating that inhibition of eIF6 expression did not affect the mechanical and thermal pain thresholds of normal mice. Compared with db/db group, PWT and TWL of E+db/db group are significantly reduced, which indicates that the eIF6 knockout can effectively relieve hypoalgesia of diabetic peripheral neuropathy mice.

Claims (10)

  1. Use of the eif6 gene for the preparation of a pharmaceutical composition for the prevention or treatment of diabetic peripheral neuropathy.
  2. 2. The use according to claim 1, wherein the prevention or treatment of diabetic peripheral neuropathy refers to the prevention or treatment of hypoalgesia in diabetic peripheral neuropathy; preferably, the pain sensation comprises mechanical pain sensation and thermal pain sensation.
  3. 3. The use according to claim 1 or 2, wherein the pharmaceutical composition comprises an inhibitor of eIF6 functional expression.
  4. 4. The use according to claim 3, wherein the inhibitor is an agent that knocks out eIF6 by CRISPR/Cas9 technology.
  5. 5. A pharmaceutical composition for preventing or treating diabetic peripheral neuropathy, wherein the pharmaceutical composition comprises an inhibitor of eIF6 functional expression.
  6. 6. The pharmaceutical composition of claim 5, wherein the inhibitor is an agent that knocks out eIF6 by CRISPR/Cas9 technology.
  7. Use of the eif6 gene for screening candidate compounds for the prevention or treatment of diabetic peripheral neuropathy.
  8. 8. The use according to claim 7, wherein the step of screening candidate compounds for the prevention or treatment of diabetic peripheral neuropathy comprises:
    in a test group, adding a test compound into a culture system of cells, and observing the expression amount and/or activity of eIF6 in the cells of the test group; in the control group, no test compound is added in the culture system of the same cells, and the expression amount and/or activity of eIF6 in the cells of the control group are observed; wherein, if the expression level and/or activity of eIF6 in the cells in the test group is lower than that in the control group, the test compound is a candidate compound for treating diabetic peripheral neuropathy having an inhibitory effect on expression and/or activity of eIF6.
  9. 9. Use of an agent for detecting eIF6 gene expression levels in the preparation of a product for diagnosing diabetic peripheral neuropathy.
  10. 10. Use according to claim 9, characterized in that the agent is selected from: a probe that specifically recognizes eIF 6; or a primer that specifically amplifies eIF 6; or an antibody or ligand that specifically binds to a protein encoded by eIF6.
CN202310886788.XA 2023-07-19 2023-07-19 Diagnosis and treatment marker for diabetic peripheral neuropathy and application thereof Pending CN116650652A (en)

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