论文部分内容阅读
目的探讨β1转化生长因子(TGF-β1)在猬迭宫绦虫(Spirometra erinacei)裂头蚴感染小鼠皮下肌肉组中的表达情况及意义。方法对采自野生王锦蛇(Elaphe carinata)的裂头蚴进行形态学观察和PCR检测。昆明小鼠80只,用数字随机表法挑选40只,雌雄各半作为实验组,剩余40只为对照组。用蛇源裂头蚴经口喂饲实验组小鼠,5条/鼠。于喂饲后第7、14、28、56天各随机剖杀10只,分别收集含有裂头蚴寄生的皮下肌肉组织,用中性甲醛固定,制作石蜡切片。HE染色后,观察裂头蚴感染病灶周围的病理变化和纤维化程度;免疫组化检测皮下肌肉中TGF-β1的含量变化。对照组不喂饲裂头蚴,检测时间及方法同实验组。结果PCR扩增获得约400 bp的目的条带。测序结果显示,该裂头蚴COX1基因与Gen Bank中的猬迭宫绦虫序列一致性为99.12%。HE染色检查,皮下肌肉中的裂头蚴被炎性囊壁所包裹,虫体与囊壁之间形成穴腔,穴腔有时出现少许的浆液或血液。囊壁最里面的区域有薄层的纤维蛋白、坏死的碎片,壁中早期以中性粒细胞浸润为主,也有嗜酸粒细胞、巨噬细胞、浆细胞等,随着病程的进展,炎性囊壁逐渐扩大,壁中以慢性炎性细胞浸润为主,囊壁周围是宿主的组织细胞,纤维结缔组织增生明显,并有不同程度的纤维化。免疫组化检查结果显示,TGF-β1的主要表达部位为裂头蚴周围的炎症反应带和纤维结缔组织增生部位。TGF-β1相对表达量在感染后逐渐增高,于感染第28天达峰值(0.654 5±0.045 5),第56天时明显下降。在感染后第7~56天,实验组的TGF-β1水平为(0.502 6±0.008 2)~(0.346 8±0.030 4)与同期对照组的(0.270 0±0.001 6)~(0.274 0±0.005 1)比较,差异均有统计学意义(P<0.05)。结论小鼠感染猬迭宫绦虫裂头蚴早、中期,TGF-β1表达水平均提高,其免疫抑制作用不利于清除和控制皮下肌肉组织中的裂头蚴。
Objective To investigate the expression of β1 transforming growth factor (TGF-β1) in the subcutaneous muscle of Spirometra erinacei infected mice and its significance. Methods Morphological observation and PCR detection of the spider larvae collected from the wild Elaphe carinata were carried out. A total of 80 Kunming mice were randomly selected by random number table method. The male and female were selected as the experimental group and the remaining 40 as the control group. The experimental group mice were fed with snakehead spider larvae, 5 / mouse. On the 7th, 14th, 28th, and 56th days after feeding, 10 rabbits were randomly killed. The subcutaneous muscle tissues containing parasitic sporocysts were collected and fixed with neutral formaldehyde to produce paraffin sections. After HE staining, the pathological changes and the degree of fibrosis were observed around the infection of the splenicculosis. The changes of TGF-β1 in subcutaneous muscles were detected by immunohistochemistry. The control group was not fed with sporozoite, the test time and method were the same as the experimental group. Results The target band of about 400 bp was obtained by PCR. Sequencing results showed that there was 99.12% identity between the COX1 gene of Cracked nematodes and that of Genome Hedgehog. HE staining, subcutaneous muscle in the cercariae are enveloped by the inflammatory wall, parasites and the formation of pockets between the wall, cavity sometimes appear a little slurry or blood. The innermost area of the wall has a thin layer of fibrin, necrotic fragments, the wall mainly in the early neutrophil infiltration, but also eosinophils, macrophages, plasma cells, etc., as the course of disease, inflammation The wall of the cyst is gradually enlarged. The wall is mainly infiltrated by chronic inflammatory cells. The cytoplasm around the cyst is the host cell of the host. The fibrous connective tissue has obvious hyperplasia and fibrosis with different degrees. The results of immunohistochemistry showed that the main expression sites of TGF-β1 were the inflammatory reaction zone around the sporozoite and the fibrous connective tissue hyperplasia. The relative expression of TGF-β1 increased gradually after infection and peaked on the 28th day of infection (0.654 5 ± 0.045 5), and decreased significantly on the 56th day. The levels of TGF-β1 in the experimental group were (0.502 6 ± 0.008 2) ~ (0.346 8 ± 0.030 4) and (0.270 ± 0.001 6) ~ (0.274 ± 0.005) 1), the differences were statistically significant (P <0.05). Conclusions The expression of TGF-β1 in early stage, mid stage and early stage of the metacercariae infected with hedgehog was increased. The immunosuppressive effect was not conducive to the clearance and control of the sporozoite in subcutaneous muscle tissue.