Novel diagnosis and treatment marker sKLB for sepsis intestinal mucous barrier damage and application of novel diagnosis and treatment marker sKLB

Abstract

The invention provides application of a novel biomarker sKLB of sepsis intestinal mucous barrier damage in detection, diagnosis and treatment of sepsis. Clinical sample exploration proves that the sKLB has good correlation with sepsis in people, so that a new diagnosis and treatment marker is provided for clinical diagnosis and treatment of sepsis, and the application has important clinical significance.

Description

TECHNICAL FIELD

[0001] The present application relates to a novel diagnostic and therapeutic marker for intestinal mucus barrier damage in sepsis. BACKGROUND

[0002] Sepsis is a complex clinical syndrome, usually accompanied by systemic inflammatory response and multiple organ failure. Current treatment of sepsis is limited to supportive options, including fluid resuscitation, vasoactive drugs and mechanical ventilation, but these methods cannot directly target the pathophysiological mechanism of sepsis. Therefore, the morbidity and mortality of sepsis are still high; although recent studies have shown that the pathophysiology of sepsis involves inflammation, microvascular dysfunction, oxidative stress and apoptosis, the exact mechanism is still unclear.

[0003] KLB belongs to the Klotho family of long-lived proteins and is widely expressed in various cells. As a high-affinity FGF receptor, KLB forms a zipper-like FGF-FGFR-KLB ternary complex, promotes fatty acid oxidation, improves insulin sensitivity, regulates blood glucose levels, and plays a key role in maintaining metabolic balance. KLB contains two subtypes, membrane KLB and soluble KLB (sKLB), and through in-depth research on the mechanism of sepsis, the present application finds that sKLB as a potential new diagnostic and therapeutic marker can improve the efficiency and accuracy of sepsis diagnosis and treatment, and has important clinical value. SUMMARY

[0004] The first object of the present application is to provide a marker for the diagnosis and treatment of sepsis, which is sKLB.

[0005] The second object of the present application is to provide an application of a marker for the diagnosis and treatment of sepsis.

[0006] To achieve the above technical objects, the present application studies the expression of sKLB in the serum of sepsis population and healthy control population, and finds that the expression of sKLB in the serum of sepsis population is reduced.

[0007] Further, the present application studies the expression of sKLB in the feces of sepsis population and healthy control population, and finds that the expression of sKLB in the feces of sepsis population is also reduced.

[0008] According to the above research results, sKLB can be preliminarily used as a marker for sepsis for the diagnosis or prognosis prediction of sepsis.

[0009] The present application provides the application of sKLB in the preparation of sepsis diagnosis or prognosis prediction products.

[0010] Further, the present application provides the use of the reagent for detecting sKLB expression in the preparation of a product for diagnosing or predicting prognosis of sepsis.

[0011] The kit comprises an antibody against a certain peptide segment when detecting the expression amount of sKLB protein using Western blot, ELISA, chemiluminescence or immunofluorescence.

[0012] The present application also provides the use of sKLB in screening and preparing a pharmaceutical composition for treating sepsis.

[0013] The present application also provides a pharmaceutical composition for treating sepsis, which comprises a reagent or compound for promoting sKLB expression.

[0014] The pharmaceutical composition of the present application can be administered alone or together with other drugs.

[0015] The pharmaceutical composition of the present application can be prepared into various formulations as needed. These include, but are not limited to, tablets, solutions, granules, patches, ointments, capsules, aerosols or suppositories for transdermal, mucosal, nasal, buccal, sublingual or oral use.

[0016] The administration route of the pharmaceutical composition of the present application is not limited as long as it can exert the desired therapeutic or prophylactic effect. In some cases, it can be administered systemically, and in some cases, it can be administered locally.

[0017] The dose of the pharmaceutical composition of the present application is not limited as long as the desired therapeutic or prophylactic effect is obtained, and can be appropriately determined according to symptoms, gender, age, etc.

[0018] The dose of the therapeutic or prophylactic pharmaceutical composition of the present application can be determined using, for example, the therapeutic or prophylactic effect on the disease as an index.

[0019] The present application finds a marker for diagnosing and treating sepsis, and provides a new use of sKLB in preparing a method or product for diagnosing or predicting prognosis of sepsis. It provides a new target for the diagnosis and treatment of clinical sepsis. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 For the analysis of the expression amount of sKLB in the serum of the sepsis population, **P<0.01.

[0021] Figure 2 For the analysis of the expression amount of sKLB in the feces of the sepsis population, *P<0.05. DETAILED DESCRIPTION

[0022] The application will be further described by specific embodiments below, and the following embodiments are specific embodiments of the application, but the embodiments of the application are not limited by the following embodiments. The application can also be implemented or applied by other different specific embodiments, and various modifications or changes can be made to the details in the specification based on different views and applications without departing from the spirit of the application.

[0023] Example 1: ELISA method for detecting KLB expression in serum of sepsis and control population respectively

[0024] According to the reagent instruction of Human Beta-klotho ELISA Kit of American Signalway Antibody.

[0025] (1) Prepare all reagents, samples and standards.

[0026] (2) Add 100 μl of standard, blank control or sample to each well, and seal the plate with sealing film. Incubate at 37°C for 2 hours.

[0027] (3) Remove the liquid in each well without washing. Add 100 μl of prepared detection reagent A to each well, and cover with sealing film. Incubate at 37°C for 1 hour.

[0028] (4) Blow and wash each well for 3 times.

[0029] (5) Add 100 μl of prepared detection reagent B to each well, and cover with new sealing film. Incubate at 37°C for 1 hour.

[0030] (6) Blow and wash each well for 5 times.

[0031] (7) Add 90 μl of substrate solution to each well, and cover with new sealing film. Incubate at 37°C for 10-20 minutes in the dark.

[0032] (8) Add 50 μl of stop solution to each well, and immediately read at 450 nm.

[0033] Example 2: ELISA method for detecting KLB expression in feces of sepsis and control population respectively

[0034] (1) Pretreatment of fecal sample: weigh an appropriate amount of fecal sample in sterile normal saline or PBS buffer, homogenize thoroughly and centrifuge, take the supernatant and filter through a sterile filter membrane to obtain clear fecal filtrate.

[0035] (2) Detection according to the above reagent instruction.

[0036] Statistical method

[0037] The results were analyzed using Graphpad Prism 8.0 statistical analysis software, and t test was used for comparison between samples.

[0038] Results

[0039] (1) Analysis of the expression of KLB in the serum of sepsis patients.

[0040] The experimental results show that the expression of KLB in the serum of sepsis patients is lower than that of the normal control group, and the results are shown in Figure One .

[0041] (2) Analysis of the expression of KLB in the stool of sepsis patients.

[0042] The experimental results show that the expression of KLB in the stool of sepsis patients is lower than that of the normal control group, and the results are shown in Figure Two .

Claims

1. Use of an agent for detecting sKLB expression in the preparation of a diagnostic or therapeutic, prognostic product for the intestinal mucus barrier impairment in sepsis.

2. Use according to claim 1, the agent or kit being used for the diagnostic or prognostic detection of the expression of sKLB in serum or blood samples or urine samples or fecal samples in sepsis.

3. Use of sKLB in the selection of a pharmaceutical composition for the treatment of sepsis.

4. Use of sKLB in the preparation of a pharmaceutical composition for the treatment of sepsis.

5. A pharmaceutical composition for treating sepsis, characterized by, The pharmaceutical composition comprises an agent or a compound that promotes the expression of sKLB.

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