目的 了解ADAM金属肽酶结构域23（ADAM23）在多房棘球蚴（Em）感染小鼠肝纤维化进程中的表达变化。方法 36只C57BL/6小鼠随机分为6组，每组6只，1个月假手术组、1个月Em感染组、3个月假手术组、3个月Em感染组、6个月假手术组、6个月Em感染组。Em感染组小鼠通过门静脉注射Em原头节悬液（2 000个原头节/鼠），假手术组注射等量生理盐水，分别在感染后1、3、6个月取肝组织。经苏木素-伊红（HE）和天狼星红染色，观察肝小叶结构、炎性细胞浸润、肉芽肿和胶原纤维沉积情况，免疫组化染色观察目标蛋白的表达情况。取3个月假手术组和3个月Em感染组小鼠肝组织进行蛋白质组学分析，蛋白质免疫印迹（Western blotting）和实时荧光定量PCR（RT-qPCR）检测ADAM23、α-平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原白αⅠ链（COL1A1）在各组小鼠中的表达水平变化。将人肝星状细胞接种于6孔板中（2 × 10⁵个/孔），分为si-ADAM23组、转化生长因子（TGF-β1）+ 小干扰RNA（siRNA）组、TGF-β1 + si-ADAM23组、siRNA组、对照组。si-ADAM23组加入600 ng siRNA-ADAM23，TGF-β1 + siRNA组加入10 ng TGF-β1和600 ng siRNA，TGF-β1 + si-ADAM23组加入10 ng TGF-β1和600 ng siRNA-ADAM23，siRNA组加入600 ng siRNA，对照组加入等量转染试剂。采用Western blotting和RT-qPCR检测ADAM23、α-SMA和COL1A1在人肝星状细胞中的表达情况。结果 HE染色结果显示，Em感染组小鼠肝小叶结构紊乱并伴有大量炎性细胞浸润，天狼星红染色结果显示，1、3、6个月Em感染组和假手术组小鼠肝纤维化面积分别为（2 167.00 ± 356.70）、（163.10 ± 25.41）μm²，（3 792.00 ± 596.00）、（159.40 ± 3.97）μm²，（5 436.00 ± 427.50）、（274.40 ± 38.01）μm²，差异均有统计学意义（t = 14.71、26.68、37.91，均P < 0.05）。蛋白质组学结果显示，3个月Em感染组小鼠ADAM23的蛋白丰度为9 250 ± 5 628。Western blotting结果显示，1、3、6个月Em感染组和假手术组小鼠ADAM23、α-SMA、COL1A1蛋白表达水平差异均有统计学意义（t = 13.5、15.8、26.1；4.37、6.75、15.82；5.00、8.65、21.45，均P < 0.05）。RT-qPCR检测结果显示，1、3、6个月Em感染组和假手术组小鼠adam23、acta2、col1a1的mRNA相对表达水平差异均有统计学意义（t = 5.44、6.44、9.95；4.13、7.43、25.10；4.85、22.60、39.50，均P < 0.05）。免疫组化结果显示，1、3、6个月Em感染组和假手术组小鼠ADAM23、α-SMA、COL1A1的阳性区域面积差异均有统计学意义（t = 26.68、17.55、52.07；24.98、36.70、49.47；16.24、45.48、77.45，均P < 0.05）。Spearman相关性分析结果显示，在Em感染小鼠的肝脏中，ADAM23阳性区域面积与α-SMA（r = 0.83，P < 0.05）、COL1A1（r = 0.87，P < 0.05）及天狼星红染色（r = 0.88，P < 0.05）阳性区域面积均呈正相关。RT-qPCR检测结果显示，对照组、siRNA组和si-ADAM23组adam23的mRNA相对表达水平分别为1.03 ± 0.14、0.92 ± 0.09、0.23 ± 0.04，与siRNA组相比，si-ADAM23组的表达水平降低（t = 12.23，P < 0.05）。Western blotting结果显示，siRNA组、TGF-β1 + siRNA组和TGF-β1 + si-ADAM23组ADAM23蛋白的相对表达水平分别为0.15 ± 0.01、1.20 ± 0.11、0.41 ± 0.15，siRNA组和TGF-β1 + siRNA组相比，TGF-β1 + siRNA组和TGF-β1 + si-ADAM23组相比差异均有统计学意义（t = 16.52、12.44，均P < 0.05）。siRNA组、TGF-β1 + siRNA组和TG-β1 + si-ADAM23组α-SMA蛋白的相对表达水平分别为0.31 ± 0.09、1.21 ± 0.04、0.87 ± 0.01，siRNA组和TGF-β1 + siRNA组相比，TGF-β1 + siRNA组和TGF-β1 + si-ADAM23组相比差异均有统计学意义（t = 27.85、10.67，均P < 0.05）。siRNA组、TGF-β1 + siRNA组和TGF-β1 + si-ADAM23组COL1A1蛋白的相对表达水平分别为0.61 ± 0.16、0.95 ± 0.02、0.49 ± 0.07，siRNA组和TGF-β1 + siRNA组相比，TGF-β1 + siRNA组和TGF-β1 + si-ADAM23组相比差异均有统计学意义（t = 5.77、7.96，均P < 0.05）。结论 ADAM23的表达水平随着Em感染进程不断升高，其表达与肝纤维化水平呈正相关。有望成为Em感染所致肝纤维化新的治疗靶点。

Objective To investigate the changes of ADAM metallopeptidase domain 23 (ADAM23) expression during the progression of hepatic fibrosis in mice infected with Echinococcus multilocularis (Em). Methdos Thirty-six C57BL/6 mice were randomly divided into six groups, of 6 mice each group, including the 1-month sham group, 1-month Em infection group, 3-month sham group, 3-month Em infection group, 6-month sham group, and 6-month Em infection group. Mice in the Em infection groups were injected with Em protoscolex suspensions (2 000 protoscoleces/mouse) via the portal vein, while animals in the sham groups received an equal volume of physiological saline. Mouse liver tissues were sampled 1, 3 and 6 months post-infection. Hepatic lobule structure, inflammatory cell infiltration, granuloma formation, and collagen deposition were observed by HE and Sirius red staining, and target protein expression was quantified using immunohistochemical staining. Mouse liver tissues from the 3-month sham and 3-month Em infection groups were sampled for proteomic analyses, and the translational and transcriptional expression of ADAM23, α-smooth muscle actin (α-SMA), and collagen type 1 alpha 1 chain (COL1A1) was determined in mice using Western blotting and RT-qPCR assays. Human hepatic stellate cells (HSCs) were seeded onto 6-well plates at a density of 2 × 10⁵ cells/well and divided into the si-ADAM23 group, TGF-β1 + small interfering RNA (siRNA) group, TGF-β1 + si-ADAM23 group, siRNA group, and control group. Cells were transfected with 600 ng siRNA-ADAM23 in the si-ADAM23 group, received 10 ng TGF-β1 and 600 ng siRNA cells in the TGF-β1 + siRNA group, received 10 ng TGF-β1 and 600 ng siRNA-ADAM23 in the TGF-β1 + si-ADAM23 group, received 600 ng siRNA in the siRNA group, and received an equal volume of transfection reagents in the control group. The translational and transcriptional expression of ADAM23, α-SMA, and COL1A1 was detected in HSCs using Western blotting and RT-qPCR assays. Results HE staining showed hepatic lobule structural derangements with infiltration of a large number of inflammatory cells in Em-infected mice. Sirius red staining revealed that the areas of hepatic fibrosis were (2 167.00 ± 356.70) μm², (163.10 ± 25.41) μm², (3 792.00 ± 596.00) μm², (159.40 ± 3.97) μm², (5 436.00 ± 427.50) μm², (274.40 ± 38.01) μm² in the 1, 3, and 6-month Em infection groups and corresponding sham groups, respectively, with significant differences seen between the infection and sham groups (t = 14.71, 26.68 and 37.91; all P < 0.05). Proteomic analysis showed that the ADAM23 protein abundance was 9 250 ± 5 628 of in the 3-month Em infection group. Western blotting assay showed significant differences in ADAM23, α-SMA, and COL1A1 protein expression between the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 13.5, 15.8, 26.1; 4.37, 6.75, 15.82; 5.00, 8.65 and 21.45; all P < 0.05). RT-qPCR assay quantified significant differences in the relative mRNA expression of adam23, acta2, and col1a1 between the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 5.44, 6.44, 9.95; 4.13, 7.43, 25.10; 4.85, 22.60 and 39.50; all P values < 0.05). Immunohistochemistry showed significant differences in the area of positive ADAM23, α-SMA, and COL1A1 expression the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 26.68, 17.55, 52.07; 24.98, 36.70, 49.47; 16.24, 45.48 and 77.45; all P < 0.05). Spearman correlation analysis revealed that the area of positive ADAM23 expression positively correlated with the areas of positive α-SMA (r = 0.83, P < 0.05) and COL1A1 expression (r = 0.87, P < 0.05), and Sirius red staining (r = 0.88, P < 0.05) in liver samples of Em-infected mice. RT-qPCR assay quantified that the relative adam23 mRNA expression was 1.03 ± 0.14, 0.92 ± 0.09, and 0.23 ± 0.04 in HSCs in the control, siRNA, and si-ADAM23 groups, respectively, and higher relative adam23 mRNA expression was found in the siRNA group than in the si-ADAM23 group (t = 12.23, P < 0.05). Western blotting assay showed that the relative ADAM23 protein expression was 0.15 ± 0.01, 1.20 ± 0.11, and 0.41 ± 0.15 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23 group, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 16.52 and 12.44, both P < 0.05), and the relative α-SMA protein expression was 0.31 ± 0.09, 1.21 ± 0.04, and 0.87 ± 0.01 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 27.85 and 10.67, both P values < 0.05), while the relative COL1A1 protein expression was 0.61 ± 0.16, 0.95 ± 0.02, and 0.49 ± 0.07 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23 group, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 5.77 and 7.96, both P values < 0.05). Conclusion ADAM23 expression increases with the progression of Em infections and positively correlates with the degree of hepatic fibrosis. ADAM23 is a novel potential therapeutic target for hepatic fibrosis caused by Em infections.