目的：观察肿瘤坏死因子-α(TNF-α)在内毒素诱导的葡萄膜炎(EIU)模型大鼠虹膜、睫状体组织中的表达，以及血必净注射液对EIU大鼠TNF-α表达的影响。方法：将清洁级Wistar大鼠65只随机分为3组，生理盐水注射组5只(A组)、生理盐水+内毒素注射组30只(B组)、血必净注射液+内毒素注射组30只(C组)。B组和C组大鼠采用双后足底注射大肠杆菌内毒素诱导葡萄膜炎动物模型，并在注射内毒素30 min前分别腹腔注射生理盐水(15 mL/kg)或血必净注射液(15 mL/kg)。将B组和C组大鼠在注射内毒素后分为6、12、18、24、48、72 h六个亚组，每个亚组各5只，于上述不同时间点观察大鼠眼内炎症反应，吸取房水计数炎症细胞数目，并采用HE染色观察大鼠虹膜、睫状体组织内的病理改变，免疫组织化学方法检测术后不同时间点TNF-α在虹膜、睫状体组织中的表达并分析。结果：临床观察显示，A组大鼠未出现葡萄膜炎体征，B组大鼠可见炎症反应，C组与B组相比炎症反应显著减轻(P<0.01)； 房水细胞计数显示A组房水中未见炎性细胞，C组房水细胞数量较B组显著减少(P<0.01)； HE染色显示A组无细胞浸润，B组在内毒素注射后6 h即可观察到眼部出现细胞浸润，随着时间推移浸润细胞数逐渐增加，24 h达到峰值后逐渐减少； C组在内毒素注射后6 h炎性细胞浸润不明显，24 h可见少量炎性细胞浸润，48 h炎症细胞浸润几乎消失。免疫组化染色显示B组大鼠虹膜、睫状体组织中TNF-α表达高于A组(P<0.01)，且TNF-α在C组的表达相较B组减弱(P<0.01)。结论：TNF-α在EIU模型大鼠中表达增高，其平均光密度比值与炎症程度呈正相关，且血必净注射液可通过降低TNF-α的水平减轻内毒素诱导的葡萄膜炎的炎症反应。

AIM: To evaluate the expression of tumor necrosis factor-α(TNF-α)in the iris and ciliary body of Wistar rats in the endotoxin-induced uveitis(EIU), and the effect of Xuebijing injection on its expression.METHODS:A total of 65 Wistar rats were randomly divided into three groups: Group A(normal saline, n=5), Group B(normal saline+endotoxin-injected, n=30), and Group C(Xuebijing+endotoxin-injected, n=30). The EIU model was induced in Wistar rats of the groups B and C by injecting LPS into the plantar surfaces of the hind feet, and normal saline(15 mL/kg)or Xuebijing(15 mL/kg)were intraperitoneally administered 30 min before LPS administration. The rats of the groups B and C were further divided into 6 subgroups after LPS injection, including 6, 12, 18, 24, 48, and 72 h subgroups, with 5 rats in each group. Furthermore, the intraocular inflammation of the rats was observed at each time above, the number of infiltrating cells in the aqueous humor was counted, and the pathological changes were observed in the iris and ciliary body of rats using hematoxylin and eosin(HE)staining. TNF-α expression in iris and ciliary tissue at different postoperative time points was evaluated using immunohistochemistry.RESULTS: Clinical observations indicated no signs of uveitis in the group A, signs of uveitis were observed in the group B. Both iris symptoms and damage were significantly reduced in the group C compared to the group B(P<0.01). Cell counts in the aqueous humor revealed no inflammatory cells in the group A, while the number of aqueous humor cells in the group C was significantly reduced compared to Group B(P<0.01). HE staining revealed no cellular infiltration in the group A. In the group B, some cellular infiltration was observed in the eyes at 6 h post-LPS exposure. The number of infiltrating cells increased over time, peaked at 24 h, and gradually declined thereafter. In the group C, cell infiltration was not obvious at 6 h, few at 24 h, and nearly disappeared by 48 h. Immunohistochemical staining showed higher TNF-α expression in the ciliary body and iris in the group B than in the group A(P<0.01). Compared to the group C, TNF-α expression in the group B was significantly upregulated following LPS injection(P<0.01).CONCLUSION:TNF-α expression was elevated in EIU rats, and there was a positive correlation between its mean optical density ratio and inflammation degree. Moreover, Xuebijing injection could alleviate inflammation response through the reduction of TNF-α levels.

山东省中医药科技项目(No.2021M085)