CN-122012697-A - Application of LncRNA EBLN3P in diagnosis and treatment of rheumatoid arthritis

Abstract

The invention provides application of LncRNA EBLN3P in diagnosis and treatment of rheumatoid arthritis, belonging to the field of rheumatic immune molecular biology. LncRNA EBLN3P can promote proliferation, migration and angiogenesis factor secretion of rheumatoid arthritis fibroblast-like synovial cells, thereby being applied to diagnosis of rheumatoid arthritis and preparation of therapeutic agent. Meanwhile, lncRNA EBLN3P can regulate and control NFIX expression through targeting miR-369-3P, and promote RA synovial membrane angiogenesis. The invention proves that LncRNA EBLN3P can play a spongy role, is competitively combined with miR-369-3P to lose use, further regulates and controls the expression of a corresponding target gene NFIX and promotes RA angiogenesis, thereby proving that the invention can be used for targeted therapy of RA synovial angiogenesis.

Inventors

• WANG YUAN
• Fang Mali
• LI TENG
• LIU FEIFEI
• Cong Chengzhi
• LIU JIAN
• HUANG CHUANBING
• SUN MENGYU
• WU AIHONG
• JIN CHAO
• ZHOU YIXUAN

Assignees

• 安徽中医药大学第一附属医院(安徽省中医院)

Dates

Publication Date

20260512

Application Date

20260213

Claims (5)

1. 1. A RA mark long-chain non-coding RNA, namely LncRNA EBLN3P, can promote proliferation and migration of rheumatoid arthritis fibroblast-like synovial cells and secretion of angiogenic factors, thereby being applied to diagnosis of rheumatoid arthritis and preparation of therapeutic agents.
2. 2. The application of LncRNA EBLN3P in diagnosing rheumatoid arthritis is characterized in that a preparation for detecting the expression level of LncRNA EBLN3P gene in a peripheral blood mononuclear cell sample can be applied to the preparation of a rheumatoid arthritis detection preparation, and the detection preparation comprises a primer pair with nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO. 2.
3. 3. The use of claim 2, wherein the expression level of LncRNA EBLN3P in peripheral blood mononuclear cells of rheumatoid arthritis patients is significantly increased, wherein LncRNA EBLN3P is significantly positively correlated with RA patient ESR, CRP, CCP antibody, SDS, CPRI-RA and PF scores and significantly negatively correlated with BP, GH, VT scores.
4. 4. The application of LncRNA EBLN3P in treating rheumatoid arthritis is characterized in that a preparation containing small interfering RNA for inhibiting the expression of LncRNA EBLN3P gene can be applied to the preparation of a TNF-alpha mediated rheumatoid arthritis targeting therapeutic preparation, the nucleotide sequence of small interfering RNA Linc-EBLN3P-1734 (Si 1) is shown as SEQ ID NO.3 and SEQ ID NO.4, the nucleotide sequence of small interfering RNA Linc-EBLN3P-2228 (Si 2) is shown as SEQ ID NO.5 and SEQ ID NO.6, and the nucleotide sequence of small interfering RNA Linc-EBLN3P-3201 (Si 3) is shown as SEQ ID NO.7 and SEQ ID NO. 8.
5. 5. The application of LncRNA EBLN3P as miRNA molecular sponge in regulating and controlling the non-therapeutic purpose of inhibiting the function of specific binding sequence miR-369-3P is characterized in that the miR-369-3P can promote NFIX gene expression after being inhibited, so as to activate JAK/STAT signal pathway and promote RA synovial angiogenesis.

Description

Application of LncRNA EBLN3P in diagnosis and treatment of rheumatoid arthritis Technical Field The invention belongs to the field of rheumatic immune molecular biology, and particularly relates to application of LncRNA EBLN3P in diagnosis and treatment of rheumatoid arthritis. Background Rheumatoid arthritis (rheumatoid arthritis, RA) is a multifactorial chronic autoimmune disease of unknown etiology that can lead to progressive joint destruction. Angiogenesis with synovial pannus formation and inflammation is the basis for RA joint destruction. Angiogenesis plays an important role in the generation of synovial pannus and is a key factor in the generation and maintenance of pannus. In an inflammatory environment, blood vessels first undergo a period of high inflammation followed by a period of dramatic increase in blood vessel growth. The newly formed blood vessels provide oxygen and nutrition to the proliferating synovitis cells, further promote the progress of synovial pannus and promote joint invasion and destruction. During sustained synovial pannus formation and progressive articular cartilage erosion, RA-FLS plays a central role, and these FLS located within the pannus produce pathogenic mediators such as metalloproteases, inflammatory cytokines, etc., that exhibit invasive phenotypes and play a central role in joint inflammation and cartilage destruction. In addition, FLS is also a key participant in initiating and maintaining immune cell recruitment, and can induce migration and differentiation of other cells within the RA synovium, thereby accelerating disease progression. And the in vitro invasive properties of FLS are related to imaging joint damage in RA patients. Different angiogenic factors promote disease progression, making anti-angiogenic therapy the focus of RA therapy. Research shows that non-coding RNAs (ncrnas) such as long-chain non-coding RNAs (lncrnas) and micrornas (mirnas) account for more than 98% of human genome play an important role in gene expression and regulation. Wherein long non-coding RNA (LncRNA) belongs to a non-coding RNA family, and can regulate the expression of target genes at multiple levels. Abnormal expression of LncRNA and its dysfunction are closely related to the occurrence of various diseases in humans, and it is possible to regulate gene expression by exerting the action of "regulatory factors", one of which is "sponge" RNA acting as miRNA, which effectively reduces its expression in cells by competitively binding to miRNA, thus inhibiting their ability to target mRNA, and this way of LncRNA-miRNA-mRNA (i.e. competitive ceRNA) interaction is critical in various biological processes. Studies have shown that the presence of abnormally expressed LncRNAs, miRNAs and mRNAs in blood and synovial tissue of RA patients, between these abnormally expressed LncRNAs, miRNAs and mRNAs, can modulate a range of biological processes (e.g. cell growth cycle, proliferation, migration, etc.) of FLS cells by this interaction to affect cytokine production, mediating disease progression. Disclosure of Invention First, the invention provides a novel RA marker long-chain non-coding RNA and application thereof in rheumatoid arthritis. The proposed novel RA marker long-chain non-coding RNA, namely LncRNA EBLN3P (GenBank sequence number: NR_ 036592.1), can promote proliferation and migration of rheumatoid arthritis fibroblast-like synovial cells and secretion of angiogenic factors, and can be applied to diagnosis of rheumatoid arthritis and preparation of therapeutic agents. Then, the invention also provides application of a preparation for detecting the expression quantity of LncRNA EBLN3P genes in peripheral blood mononuclear cell samples in preparation of rheumatoid arthritis detection preparations, wherein the preparation for detecting the expression quantity of LncRNA EBLN3P genes comprises primer pairs with nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO. 2. Experiments prove that the expression quantity of LncRNA EBLN3P in peripheral blood mononuclear cells of rheumatoid arthritis patients is obviously increased, and LncRNA EBLN3P is obviously positively correlated with the scores of RA patients ESR, CRP, CCP antibody, SDS, CPRI-RA and PF, and is obviously negatively correlated with the scores of BP, GH and VT. The invention further provides application of the specific small interfering RNA of the LncRNA EBLN3P gene in preparing a TNF-alpha mediated rheumatoid arthritis targeted therapeutic agent, or application of the small interfering RNA for inhibiting the expression of the LncRNA EBLN3P gene in preparing a rheumatoid arthritis targeted therapeutic agent. The invention designs and synthesizes specific over-expression sequences pcDNA3.1-LncRNA EBLN3P and a small interference sequence Si-LncRNA EBLN3P aiming at the full-length sequence of LncRNA EBLN3P, which are used for transfecting RA-FLS to ensure that the LncRNA EBLN3P is over-expressed and small interfered in ce