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利用高脂饲料和异种血清诱导建立肝纤维化模型,检测炎性因子的表达情况。将BALB/c小鼠随机分为正常小鼠组、高脂饲料饮食组、腹腔注射异种血清组、高脂饲料联合腹腔异种血清注射组。6周后,比较4组小鼠体重以及肝重指数,收集肝脏标本经HE和Masson染色后光镜观察肝组织的结构变化以及胶原纤维状况,并提取肝组织的总RNA,RTPCR检测白介素-17A(IL-17A)、基质金属蛋白酶抑制剂(TIMP)、Ⅰ型胶原蛋白(COLL)、肿瘤坏死因子-α(TNF-α)等基因mRNA水平的改变。结果显示与正常对照组体重及肝重指数(26.7±2.5g,5.24±0.81)相比显示,高脂饲料饮食组(30.4±3.7g,5.44±0.32)和高脂饲料联合腹腔异种血清注射组(30.3±1.8g,6.51±1.13)体重及肝重指数明显升高(P<0.05);HE和Masson染色显微镜下也观察到肝细胞脂肪变性,大量的胶原纤维产生;RT-PCR检测TIMP,IL-17A,COLL以及TNF-α的mRNA,相比于正常对照组,实验组肝组织mRNA表达均有升高且具有统计学意义(P<0.05)。本实验应用高脂和异种血清联合处理建立了小鼠肝纤维化模型,并发现炎性因子IL-17A及TNF-α参与了小鼠肝纤维化的形成过程。
The use of high-fat diet and heterologous serum induced liver fibrosis model was established to detect the expression of inflammatory cytokines. The BALB / c mice were randomly divided into normal mice group, high fat diet group, intraperitoneal injection of heterologous serum group and high fat diet combined with intraperitoneal injection of heterologous serum. Six weeks later, the body weight and liver index of four groups were compared. The liver samples were collected and stained with HE and Masson to observe the changes of liver structure and collagen fibers. The total RNA of liver tissue was extracted. The expression of interleukin-17A (IL-17A), tissue inhibitor of metalloproteinase (TIMP), collagen type Ⅰ (COLL) and tumor necrosis factor-α (TNF-α) The results showed that compared with normal control group (26.7 ± 2.5g, 5.24 ± 0.81), high fat diet group (30.4 ± 3.7g, 5.44 ± 0.32) and high fat diet group combined with intraperitoneal injection (30.3 ± 1.8g, 6.51 ± 1.13) and liver weight index (P <0.05). Steatosis of hepatocytes and massive collagen fibers were also observed under HE and Masson staining. TIMP, Compared with normal control group, the mRNA expression of IL-17A, COLL and TNF-αin the experimental group were increased and statistically significant (P <0.05). In this study, the model of hepatic fibrosis was established by the combination of high-fat and heterogeneous serum. The inflammatory factors IL-17A and TNF-α were found to be involved in the formation of hepatic fibrosis in mice.