基于“久病入络”理论的当归饮子防治荨麻疹PIP2/IP3/DAG信号通路的分子研究

Urticaria is a common clinical allergic skin disease, with high morbidity and recurrence rate, and belongs to "collateral disease" of traditional Chinese medicine.As research of urticaria pathological mechanism goes deeper, we find that mastocyte degranulation plays an important role in the pathogenesis of urticaria, and PIP2/IP3/DAG signal pathway is one of the important mechanisms on regulation of mastocyte degranulation, and closely related to onset of urticaria.Pre-experiment of research group has found that Danggui yinzi inhibits mastocyte degranulation of rash site of mouse skin, reduces both ear swelling on sensitized mouce, regulates T-cell function, and inhibits expression of RANTES,MCP-1 and MIP-1.Clinical observation has found that Danggui yinzi reduces itching, decreases the level of serum inflammatory mediators IL-4,TNF-α of urticaria patients. We speculate that Danggui yinzi probably functions regulation of PIP2/IP3/DAG signal pathway to inhibit mastocyte degranulation to prevent and treat urticaria.This project uses modern techniques (Fluorescence Quantitation PCR and Western Blot) and molecular biological knowledge (PIP2/IP3/DAG) to interpret material basis of the basic theory of TCM (long-term illness penetrating into collateral) ,and from a brand-new perspective to explore the modern scientific mechanism of Danggui yinzi on prevention and treatment of urticaria.

荨麻疹是临床上常见的过敏性皮肤病，发病率及复发率高，属中医"络病"范畴。随着对荨麻疹病理机制的深入研究，发现肥大细胞脱颗粒在荨麻疹的发病过程中起着重要作用，PIP2/IP3/DAG信号通路是调控肥大细胞脱颗粒的重要机制之一，与荨麻疹发作密切相关。课题组前期应用"久病入络"理论发现当归饮子可抑制小鼠皮疹部位肥大细胞脱颗粒，减轻致敏小鼠左右耳肿胀度，调节T细胞功能，抑制RANTES、MCP-1，3和MIP-1的表达；临床观察发现当归饮子可减轻荨麻疹患者瘙痒症状并防止复发，降低血清炎症介质IL-4、TNF-α水平；我们推测当归饮子可能是通过调控PIP2/IP3/DAG信号通路抑制肥大细胞脱颗粒而起防治荨麻疹的作用。因此，我们采用现代技术方法（荧光定量PCR和Western Blot）和分子生物学知识（PIP2/IP3/DAG）来阐释中医基础理论（久病入络）的物质基础，从全新的角度探索当归饮子。

荨麻疹是临床上常见的过敏性皮肤病，发病率及复发率高，属中医“络病”范畴。随着对荨麻疹病理机制的深入研究，发现肥大细胞脱颗粒在荨麻疹的发病过程中起着重要作用，PIP2/IP3/DAG信号通路是调控肥大细胞脱颗粒的重要机制之一，与荨麻疹发作密切相关。课题组前期应用“久病入络”理论发现当归饮子可抑制小鼠皮疹部位肥大细胞脱颗粒，减轻致敏小鼠左右耳肿胀度，调节T细胞功能，抑制RANTES、MCP-1，3和MIP-1的表达；临床观察发现当归饮子可减轻荨麻疹患者瘙痒症状并防止复发，降低血清炎症介质IL-4、TNF-α水平；肥大细胞脱颗粒是荨麻疹发作机制的始动环节，调控PIP2/IP3/DAG信号通路抑制肥大细胞脱颗粒是防治荨麻疹的有效途径。因此课题采用现代技术方法（荧光定量PCR和Western Blot）和分子生物学知识（PIP2/IP3/DAG）来阐释中医基础理论（久病入络）的物质基础，从全新的角度探索当归饮子。.本项目通过观察当归饮子对荨麻疹动物模型肥大细胞脱颗粒、皮损组织PIP2mRNA、PKCmRNA、IP3蛋白表达的影响，血清总lgE、IFN-γ、IL-4水平含量及组织mcp-1蛋白表达、EotaxinmRNA的、对模型动物血清组胺和5-HT的影响，掌握当归饮子通过PIP2/IP3/DAG信号转导通路抑制肥大细胞脱颗粒对过敏介质的影响。本研究采用的当归饮子高、中、低剂量组对荨麻疹动物模型干预，结果表明动物血清Calcineurin、PIP2、 IP3 含量均比模型组降低，肥大细胞脱颗粒百分数均有降低，且肥大细胞脱颗粒程度均有明显降低。本研究表明PIP2 /IP3 /DAG 信号通路中的 PIP2、IP3、DAG、Ca2 +、PKC 等信号因子在肥大细胞脱颗粒导致荨麻疹发病过程中发挥重要作用，调控 PIP2 /IP3 /DAG 信号通路可有效抑制肥大细胞脱颗粒，是防治荨麻疹发作的有效途径。