Application of blood coagulation factor X as biomarker influencing abdominal fat percentage of chicken

Abstract

The invention relates to the technical field of biomarkers, in particular to application of a blood coagulation factor X as a biomarker influencing the abdominal fat percentage of chicken. The invention discloses a biomarker influencing the abdominal fat percentage of chicken. The biomarker is a blood coagulation factor X. Experimental data provided by the invention show that F10 shows a significant difference (Plt; P1t) between groups with high abdominal fat percentage (AFP > = 18%) and groups with low abdominal fat percentage (AFP < = 12%); 0.05), and the biochemical marker combination can be used as a specific molecular marker for the abdominal fat mass of the Gushi chicken. On the basis, a molecular marker assisted breeding system containing serum F10 concentration detection, TG level evaluation and phenotype data association analysis is established, and an innovative technical path is provided for high-quality breeding of Henan local chicken germplasm resources.

Description

Technical Field

[0001] The present invention relates to the technical field of biomarkers, and in particular to the application of coagulation factor X as a biomarker affecting the abdominal fat rate of chickens. Background Art

[0002] Poultry meat consumption ranks second in terms of meat consumption, after pork. While modern commercial chickens have significantly improved meat production efficiency and shortened the time to market through continuous genetic improvement for high growth rates and carcass yields, this has also led to a significant increase in abdominal fat. Excessive abdominal fat accumulation not only leads to skeletal abnormalities, metabolic disorders, and an increased risk of mortality, but also negatively impacts key economic traits such as carcass weight, feed conversion rate, production cycle, and meat quality.

[0003] Gushi chicken, a national geographical indication product unique to Gushi County, Henan Province, is a key dual-purpose poultry breed within the poultry germplasm resource bank. Its genetic diversity and unique production performance hold significant conservation value and development potential. Currently, a rapid serum-based marker-based detection system for abdominal fat percentage in Gushi chickens has not yet been established. Therefore, the development of novel molecular marker-assisted breeding technologies is urgently needed to advance the quality improvement of local chicken breeds.

[0004] Abdominal fat weight (AFW) and abdominal fat percentage (AFP), core indicators of fat deposition in chickens, have significant heritabilities of 0.62 and 0.24, respectively, indicating that genetic selection is an effective means of regulating abdominal fat traits. Traditional phenotypic-based breeding strategies require measuring abdominal fat at terminal slaughter, resulting in lengthy generation intervals (typically 2–3 years), which severely restricts breeding efficiency. Therefore, establishing a non-invasive, high-throughput early selection technology system has become an important direction for modern poultry genetic improvement.

[0005] As a central organ of lipid metabolism, the poultry liver is responsible for over 90% of de novo fatty acid synthesis. Triglycerides (TG) synthesized by hepatocytes are packaged into very-low-density lipoprotein (VLDL) particles and secreted into the bloodstream, where they are transported to adipose tissue via the bloodstream. During this process, a dynamic balance exists between lipoprotein lipase (LPL)-mediated lipolysis and lipid resynthesis within adipocytes.

[0006] It is noteworthy that this complex lipid metabolism network is precisely regulated by a variety of serum biomarkers. Among them, endocrine factors, as intertissue signaling mediators, play a core role in maintaining energy homeostasis, metabolic regulation, and reproductive function. In particular, regulatory factors secreted specifically by the liver play a key role in regulating fat metabolic pathways and abdominal fat deposition. Coagulation factor X (F10), a serine protease synthesized by the liver, has expanded its function from traditional coagulation regulation to metabolic regulation. Recent studies have revealed that F10 affects fat deposition by activating the PI3K / AKT / mTOR signaling pathway. The specific mechanisms include:

[0007] After activating the PI3K / AKT signaling axis, F10 enhances mTORC1 activity, promoting the expression of genes involved in ribosome biogenesis, protein translation, and lipid synthesis (such as SREBP1c), thereby synergizing adipogenesis and inducing muscle fiber hypertrophy. This process has been shown to be associated with metabolic reprogramming of cancer cells in the tumor microenvironment, suggesting that it may play a similar role in adipose tissue metabolism. Serum Stability and Biomarker Potential F10 is highly stable in serum, and its expression levels are significantly correlated with cardiovascular diseases (such as coronary heart disease) and tumor progression. For example, plasma F10 levels are positively correlated with carotid intima-media thickness (CIMT), suggesting that it may serve as a potential marker for atherosclerosis. However, there are currently no reports of F10 as a serum marker of abdominal fat in poultry. Summary of the Invention

[0008] The purpose of the present invention is to provide an application of coagulation factor X as a biomarker affecting the abdominal fat rate of chickens.

[0009] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0010] The present invention provides a biomarker affecting the abdominal fat rate of chickens, wherein the biomarker is coagulation factor X.

[0011] Preferably, the chicken breed is Gushi chicken.

[0012] Preferably, the content of the coagulation factor X is positively correlated with the abdominal fat rate of the chicken.

[0013] The present invention also provides the use of the biomarker in preparing a chicken abdominal fat rate detection reagent.

[0014] Preferably, the detection reagent is a kit.

[0015] The present invention also provides a kit for detecting the abdominal fat rate of chickens, comprising a reagent for detecting the biomarker.

[0016] The present invention also provides the application of the biomarker in the breeding of chicken varieties with low abdominal fat rate.

[0017] The present invention also provides a method for breeding chickens with low abdominal fat rates, which characterizes the level of chicken abdominal fat rates by measuring the content of coagulation factor X in chicken serum, thereby achieving the purpose of breeding chickens with low abdominal fat rates.

[0018] Preferably, the chicken breed is Gushi chicken.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The experimental data provided by this paper show that the F10 index combination showed significant differences (P < 0.05) between groups with high abdominal fat rates (AFP ≥ 18%) and low abdominal fat rates (AFP ≤ 12%). This biochemical marker combination can serve as a specific molecular marker for abdominal fat mass in Gushi chickens. Based on this, the present invention established a molecular marker-assisted breeding system that includes serum F10 concentration detection, TG level assessment, and phenotypic data association analysis, providing an innovative technical path for the high-quality breeding of Henan local chicken germplasm resources. DETAILED DESCRIPTION

[0021] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0022] Example 1

[0023] 1. Test materials:

[0024] 1. Experimental animals and samples

[0025] The experimental subjects (8-week-old Gushi roosters) were obtained from the Yuanyang Germplasm Resource Farm of Henan Agricultural University and were fed and watered ad libitum under the same feeding and management conditions. Sera from 16 laboratory-stored 43-week-old Gushi hens with high and low abdominal fat percentages were also collected.

[0026] 2 Reagents

[0027] AAV-EGFP (adeno-associated virus vector carrying fluorescent protein recombinant virus) and AAV-F10 (chicken coagulation factor F10) were purchased from Hanheng Biotechnology (Shanghai) Co., Ltd.; TG-linked immunosorbent assay kit and F10 enzyme-linked immunosorbent assay kit were purchased from Shanghai ELISA Biotechnology Co., Ltd.

[0028] 3 Main instruments

[0029] Centrifuge, electronic scale, blood collection needle, vacuum blood collection tube

[0030] 2. Test methods

[0031] 1. Selection of experimental subjects

[0032] Twelve 8-week-old Gushi roosters were randomly divided into experimental group and control group (6 in each group). AAV-F10 and AAV-EGFP (4×10 11 viruses / individual).

[0033] 2. Feeding methods

[0034] The chickens were housed in a laboratory shed at Henan Agricultural University and provided with free access to feed and water under the same conditions. The chickens were fed a diet of 12.40 MJ / kg energy, 20.0% protein, 6.0% crude fiber, and 8.0% crude ash.

[0035] 3. Slaughter standards for experimental individuals

[0036] The slaughter experiment was conducted in the laboratory of Henan Agricultural University. The chickens were weighed weekly and slaughtered when there was a significant difference in weight between the experimental group and the control group. After slaughter, abdominal fat weight was measured and abdominal fat percentage was calculated.

[0037] Abdominal fat percentage = (abdominal fat weight + extra-muscular fat weight) / total eviscerated weight × 100%

[0038] 4. Determination of serum biochemical indicators

[0039] The operation was performed according to the instructions of the chicken TG and F10 enzyme-linked immunosorbent assay (ELISA) kit, and then the absorbance (OD) value was measured at a wavelength of 450 nm using a microplate reader. The TG and F10 contents were calculated after drawing a standard curve.

[0040] 5. Results Statistics

[0041] Table 1 Comparison of biochemical indicators in serum

[0042]

[0043] Table 2 Statistics of body weight changes after injection of adeno-associated virus

[0044]

[0045] Table 3 Comparison of body fat characteristics of Gushi chickens after slaughter

[0046]

[0047] 6 Verification by other groups

[0048] Table 443 F10 content in serum of Gushi chickens at week 3

[0049]

[0050] 3. Results Analysis

[0051] As shown in Table 1, after in situ injection of AAV-F10 / AAV-EGFP into chicken livers, serum F10 levels in the experimental group increased significantly. Therefore, F10 was successfully overexpressed in the chicken livers, and secretion of its encoded protein was significantly increased. Furthermore, the significant increase in circulating triglyceride levels provides ample substrate for adipocytes. Triglycerides are then broken down by LPL into fatty acids, which, when taken up by adipocytes, form more and larger lipid droplets.

[0052] Twelve chickens were slaughtered and their body weight and abdominal fat mass were measured. The average body weight of the experimental group (Table 2) was significantly higher than that of the control group. Under consistent feeding conditions, feed, and drinking water, the experimental group demonstrated higher feed conversion rates and body weight gain, indicating that F10 promoted weight gain in the chickens. Table 3 also shows a higher abdominal fat percentage in the experimental group (P < 0.05), indicating that low circulating F10 levels in Gushi chickens are significantly associated with lower abdominal fat percentages.

[0053] Subsequently, the F10 content was detected in the serum of a group of 43-week-old Gushi hens. The results showed that there were significant differences in F10 between chicken groups with high and low abdominal fat rates, and the function was consistent with the previous experiments, indicating that there was a positive correlation between F10 content and chicken abdominal fat rate.

[0054] In summary, F10 can be used as a serum marker to predict abdominal fat percentage in chickens. This allows for selection of chickens based on their F10 serum levels to identify superior breeds. Specifically, individuals with low serum F10 levels can be selected to reduce abdominal fat percentage and improve economic performance, while individuals with high serum F10 levels can be eliminated.

[0055] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A biomarker affecting chicken abdominal fat rate, characterized in that: The biomarker is coagulation factor X.

2. The biomarker according to claim 1, characterized in that The chicken breed is Gushi chicken.

3. The biomarker according to claim 1, characterized in that The content of the coagulation factor X is positively correlated with the abdominal fat rate of the chicken.

4. Use of the biomarker according to any one of claims 1 to 3 in the preparation of a reagent for detecting chicken abdominal fat rate.

5. The use according to claim 4, characterized in that The detection reagent is a kit.

6. A kit for detecting abdominal fat percentage in chickens, comprising a reagent for detecting the biomarker according to any one of claims 1 to 3.

7. Use of the biomarker according to any one of claims 1 to 3 in the breeding of chicken breeds with low abdominal fat rate.

8. A method for breeding chickens with low abdominal fat content, characterized in that: By measuring the content of coagulation factor X in chicken serum, the abdominal fat rate of chickens can be characterized to achieve the purpose of breeding chickens with low abdominal fat rate.

9. The method according to claim 8, characterized in that The chicken breed is Gushi chicken.

Images