阅读说明：本技术 一种以角蛋白1作为预测和检测人体组织炎症的生物标志物的方法及其应用 (Method for predicting and detecting human tissue inflammation by using keratin 1 and application thereof ) 是由 殷卫海 张铭超 李雨嘉 于 2018-06-22 设计创作，主要内容包括：本发明提供了一种基于检测皮肤角蛋白1的变化,从而预测或检测人体组织炎症的方法。本发明以角蛋白1作为预测和检测人体组织炎症的生物标志物的方法,包括以下步骤：(1)检测受试者的皮肤组织中的角蛋白1的含量、结构中的至少一种；(2)检测健康人群皮肤组织中的角蛋白1的含量、结构中的至少一种；(3)将步骤(1)和步骤(2)中获得的角蛋白1的含量、结构相对比,计算相应的变化率；(4)根据所述变化率,诊断或预测所述受试者的组织炎症严重程度。皮肤在受到因为外界刺激而造成的炎症反应后,其角蛋白1结构或含量会发生变化,这种变化与组织炎症的严重程度呈正相关。(The invention provides a method for predicting or detecting human tissue inflammation based on detecting changes in skin keratin 1. The invention relates to a method for predicting and detecting human tissue inflammation biomarker by using keratin 1, which comprises the following steps: (1) detecting at least one of keratin 1 content, structure in skin tissue of a subject; (2) detecting at least one of the content and the structure of keratin 1 in skin tissues of healthy people; (3) comparing the content and the structure of the keratin 1 obtained in the step (1) with those obtained in the step (2), and calculating the corresponding change rate; (4) diagnosing or predicting the severity of tissue inflammation in the subject based on the rate of change. After the skin is subjected to inflammatory reaction caused by external stimulation, the keratin 1 structure or content of the skin can be changed, and the change is in positive correlation with the severity of tissue inflammation.)

1. A method for predicting and detecting inflammation of human tissue using keratin 1 as a biomarker, comprising the steps of:

(1) detecting at least one of keratin 1 content, structure in skin tissue of a subject;

(2) detecting at least one of the content and the structure of keratin 1 in skin tissues of healthy people;

(3) comparing the content and the structure of the keratin 1 obtained in the step (1) with those obtained in the step (2), and calculating the corresponding change rate;

(4) diagnosing or predicting the severity of tissue inflammation in the subject based on the rate of change.

2. The method as set forth in claim 1, wherein in the method for predicting tissue inflammation, the skin tissue is not yet observed to be damaged at the time point of detecting keratin 1 in the skin tissue.

3. The method of claim 1, wherein:

the tissue inflammation includes, but is not limited to, the following diseases: viral encephalitis, hepatitis, pneumonia, myocarditis, or nephritis.

4. The method of claim 1, wherein:

said keratin 1 content is increased in a positive correlation with the severity of tissue inflammation; the higher the keratin 1 fragment content, the higher the severity of tissue inflammation.

5. The method of claim 1, wherein:

based on the properties such as the concentration, distribution, and bilateral symmetry of keratin 1, the tissue inflammation type and location of the patient can be determined.

6. A method for detecting tissue inflammation for non-diagnostic therapeutic purposes, comprising the steps of:

(1) after the skin tissue is treated by the inflammatory response inducer containing a certain dose, detecting at least one of the content and the structure of keratin 1 in the skin tissue of the experimental subject after the treatment of the inflammatory response inducer;

(2) detecting at least one of keratin 1 content, structure in skin tissue of the subject not damaged by the inflammatory response;

(3) comparing the content and the structure of the keratin 1 obtained in the step (1) with those obtained in the step (2), and calculating the corresponding change rate;

(4) calculating or predicting the inflammatory injury degree of the subject according to the change rate.

7. The method of claim 6, wherein the inflammation inducing agent increases the level of inflammatory factors in the skin tissue and the inflammatory response inducing agent comprises lipopolysaccharide.

8. The method of any one of claims 1 to 6, wherein the detection of keratin 1 comprises one or more of gene level, protein structure change.

9. The method according to any one of claims 1 to 6, wherein keratin 1 further comprises a keratin 1 fragment with a deletion of a portion of the amino acid sequence.

10. The method of any one of claims 1 to 6, wherein the method of detecting keratin 1 content, structure comprises one or more of fluorescence spectroscopy, Raman spectroscopy, optical detection, or physical property detection.

11. A model for use in screening for drugs that inhibit or agonize tissue inflammation, wherein the screening model is constructed using the method of any one of claims 1 to 10.

12. Use of a model according to claim 11 in the manufacture of a medicament for the treatment of tissue inflammation.

Technical Field

The invention relates to a biomarker for detecting skin tissue inflammation, in particular to a detection method for predicting and detecting tissue inflammation by using keratin 1 and application thereof.

Background

Current methods of detecting tissue inflammation are commonly through blood tests. The accuracy of this approach and the degree of differentiation between different tissue inflammations is limited. Therefore, the method is very important for finding a biological index for real-time noninvasive rapid detection of tissue inflammation, and has great social and economic significance and clinical significance.

Disclosure of Invention

The present inventors have found that keratin 1 of skin tissue can be used as a novel biomarker for detection methods for detecting and predicting tissue inflammation. Thus, a method is proposed which can be used for detecting tissue inflammation based on keratin 1.

The present invention provides a method for diagnosing and predicting tissue inflammation, comprising the steps of:

(1) detecting at least one of keratin 1 content and keratin 1 structure in skin tissue of a subject after the skin tissue is subjected to tissue inflammation;

(2) detecting at least one of keratin 1 content, structure in skin tissue of the subject not subject to tissue inflammation;

(3) comparing the content and the structure of the keratin 1 obtained in the step (1) with those obtained in the step (2), and calculating the corresponding change rate;

(4) diagnosing or predicting the degree of tissue inflammation in the subject based on the rate of change.

In one embodiment, in the detecting tissue inflammation, the skin tissue may be judged to be damaged by a clinically common method at the time point of detecting keratin 1 in the skin tissue.

In one embodiment, in the method of predicting tissue inflammation, the skin tissue has not been able to observe an inflammatory response or is judged to be inflammatory using clinically common methods when the time point of keratin 1 in the skin tissue is detected.

In one embodiment, the tissue inflammation model includes, but is not limited to, the following diseases: viral encephalitis, hepatitis, pneumonia, myocarditis, and nephritis.

In one embodiment each tissue inflammation has its own unique concentration, distribution and bilateral symmetry of keratin 1. According to the invention, the type and the position of the tissue inflammation of the patient can be non-invasively and rapidly judged according to the properties of the keratin 1 such as concentration, distribution, bilateral symmetry and the like.

The present invention also provides a method for detecting tissue inflammation for non-diagnostic therapeutic purposes, comprising the steps of:

(1) after the skin tissue is treated by the tissue inflammation inducer containing a certain dosage, detecting at least one of the content and the structure of keratin 1 in the skin tissue of the experimental subject after the skin tissue is treated by the tissue inflammation inducer;

(2) detecting at least one of keratin 1 content and structure in skin tissue of the subject not subject to tissue inflammation;

(3) comparing the content and the structure of the keratin 1 obtained in the step (1) with those obtained in the step (2), and calculating the corresponding change rate;

(4) calculating or predicting the severity of tissue inflammation in the subject based on the rate of change.

In a specific embodiment, the subject is tested at a time point within 0 to 30 days, preferably within 7 days, more preferably within 3 days, of skin tissue being treated or affected by a tissue inflammatory condition.

In the present invention, the tissue inflammation inducing agent may increase the content of inflammatory factors in the tissue and increase the level of tissue inflammation, and the tissue inflammation inducing agent includes lipopolysaccharide.

In one embodiment, the keratin 1 comprises one or more of gene level, protein structure. The keratin 1 includes but is not limited to the full-length keratin 1, or the keratin 1 fragment with the deletion of partial amino acid sequence of the keratin 1, or the mixture of the fragment protein and the full-length keratin 1, and the protein is detected not only at the protein level, but also at the gene level, including modification levels such as phosphorylation and acetylation.

In one embodiment, the keratin 1 further comprises a protein structure change, wherein the protein structure comprises a dimeric structure formed by the keratin 1 and other proteins such as the keratin 10, wherein the keratin 1 comprises a keratin 1 fragment with a partial amino acid sequence deleted therefrom, i.e. the protein structure comprises a dimeric structure formed by the keratin 1 fragment with a partial amino acid sequence deleted therefrom and other proteins such as the keratin 10.

In the present invention, a keratin 1 fragment lacking a part of the amino acid sequence is also referred to as a keratin 1 degradation product, and is selected from a keratin 1 fragment lacking any number of amino acids between 1 and 644, and the deletion may be a deletion of the N-terminus of keratin 1, a deletion of the C-terminus of keratin 1, or a deletion of any combination of amino acids in the entire sequence of keratin 1. For example, the deletion of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 119, 108, 110, 111, 112, 111, 122, 114, 121, 122, 114, 121, 114, 113, 110, 112, 113, 110, 113, 121, 113, 121, 124. 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 235, 240, 237, 245, 240, 237, 238, 240, 244, 242, 247, 242, 241, 240, 237, 240, 237, 240, 242, 248. 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 371, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 349, 350, 351, 353, 354, 358, 357, 362, 364, 363, 368, 363, 368, 366, 363, 361, 372. 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 491, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 466, 465, 482, 467, 469, 470, 471, 494, 475, 476, 477, 480, 481, 495, 492, 495, 481, 480, 495, 481, 495, 78, 496. 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 514, 515, 516, 517, 518, 519, 520, 521, 522, 523, 524, 525, 526, 527, 528, 529, 530, 531, 532, 533, 534, 535, 536, 537, 538, 539, 540, 541, 542, 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 611, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573, 574, 575, 577, 576, 579, 580, 581, 582, 583, 585, 609, 587, 588, 589, 590, 592, 593, 594, 595, 605, 598, 615, 598, 600, 599, 602, 598, 602, 610, 612, 578, 611, 612, 607, 608, 612, 607, 608, 506, 610, 612, 160, 49, 620. 621, 622, 623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644 amino acid fragment of keratin 1.

In the present invention, the method for detecting the content and structure of keratin 1 or protein 1 degradation products comprises one or more of immunoblotting, immunohistochemistry, immunofluorescence, fluorescence spectroscopy, raman spectroscopy, optical detection, or physical property detection.

In another aspect of the present invention, there is also provided a model for screening drugs for inhibiting or stimulating tissue inflammation, wherein the screening model is constructed by using the aforementioned method of the present invention.

Each tissue inflammation has its own unique concentration, distribution and bilateral symmetry of keratin 1. According to the invention, the type of the tissue inflammation of the patient can be non-invasively and rapidly judged according to the properties of the keratin 1 such as concentration, distribution, bilateral symmetry and the like.

The invention further provides the use of the model in the manufacture of a medicament for the treatment of tissue inflammation.

Drawings

Figure 1 model of lipopolysaccharide-induced skin inflammation in mice. The keratin 1 content increases with time, the keratin 1 content increases.

Detailed Description

The present invention will be further illustrated by the following detailed description.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, "keratin 1", "KRT 1" are interchangeable and include keratin that can be detected using keratin 1 antibody recognition, either as a single substance or as a mixture; in the present invention, keratin 1 includes a keratin 1 fragment in which a partial amino acid sequence is deleted, the keratin 1 fragment in which any one of 1 to 644 amino acids is deleted from the N-terminus or C-terminus of keratin 1, or a combination thereof.

As used herein, "keratin 1 fragment lacking a portion of the amino acid sequence", "keratin 1 degradant" are interchangeable and refer to a keratin 1 fragment lacking any one of 1 to 644 amino acids, or a combination thereof, from the N-terminus or C-terminus of keratin 1.

As used herein, "tissue inflammation" includes, but is not limited to, the following diseases: viral encephalitis, hepatitis, pneumonia, myocarditis, and nephritis.