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目的 探讨热休克蛋白70抑制剂(Apoptozole, Az)联合阿苯达唑(Albendazole, ABZ)干预对多房棘球蚴感染小鼠模型病灶活性和淋巴细胞亚群的影响。方法 无菌分离多房棘球绦虫原头节，以每只小鼠腹腔注射10 000枚的剂量接种雌性Balb/c小鼠27只。在感染4周后，将27只小鼠随机分为对照组(腹腔注射1.4%DMSO)、Az组(腹腔注射4mg/kg Apoptozole)和Az+ABZ组(腹腔注射4 mg/kg Apoptozole+4 mg/kg ABZ),每组9只，每两天注射1次，连续注射6周。收集各组小鼠病灶组织样本进行石蜡包埋和HE染色，观察组织的病理学变化。采用免疫组化方法检测病灶组织中Em 18和Em 14-3-3蛋白的表达水平，评价病灶活性。分离小鼠肝脏、脾脏和腹腔淋巴细胞，采用流式细胞术检测和分析各组小鼠肝脏、脾脏和腹腔中淋巴细胞亚群的比例变化。结果 HE染色结果显示：与对照组比较，Az组和Az+ABZ组小鼠病灶组织中炎性细胞浸润比较明显。免疫组化结果显示：与对照组比较，Az组和Az+ABZ组小鼠病灶组织中Em 14-3-3和Em 18蛋白表达水平降低，差异有统计学意义(P<0.001)。流式细胞术检测结果显示：与对照组比较，Az组和Az+ABZ组小鼠肝脏NK细胞和B细胞表达比例升高，差异有统计学意义(P<0.01)。与对照组比较，Az+ABZ组小鼠肝脏T细胞表达比例降低，差异有统计学意义(P<0.05)。与对照组比较，Az组小鼠肝脏B细胞表达比例升高，差异有统计学意义(P<0.01)。与对照组比较，Az组和Az+ABZ组小鼠脾脏NK细胞表达比列有升高的趋势，T细胞表达比例有下降的趋势(P>0.05)。与对照组比较，Az组小鼠脾脏B细胞表达比例升高，差异有统计学意义(P<0.05)。与对照组比较，Az组和Az+ABZ组小鼠腹腔NK细胞和B细胞表达比例有升高的趋势，T细胞和CD8~+T细胞表达比例有下降的趋势(P>0.05)。与Az组比较，肝脏，脾脏，腹腔淋巴细胞中的NK细胞都有上调的趋势。结论 热休克蛋白70抑制剂Apoptozole联合阿苯达唑干预多房棘球蚴感染小鼠模型，能够抑制病灶活性，促进NK细胞向肝脏中的募集，介导对多房棘球蚴病灶的抑制。
Abstract：Objective To explore the effects of heat shock protein 70(Hsp70) inhibitor(Apoptozole, Az) combined with albendazole(ABZ) intervention on the lesion activity and lymphocyte subsets in a Echinococcus multilocularis(E.m) infectionmouse model. Methods Isolated the protoscolex(PSC) of E.m under aseptic conditions, and 27 female Balb/c mice were injected intraperitoneally with a dose of 10 000 PSC per mouse. After 4 weeks of infection, 27 mice were randomly divided into a solvent control group(intraperitoneal injection of 1.4% DMSO), an Az group(intraperitoneal injection of 4 mg/kg Az), and an Az+ABZ group(intraperitoneal injection of 4 mg/kg Az+4 mg/kg ABZ), with 9 mice in each group. The mice were injected every 2 days for 6 weeks. Liver and lesion tissue samples from each group mice were collected for paraffin embedding and HE staining to observe the pathological changes in the tissues. Immunohistochemistry was used to detect the expression levels of Em 18 and Em 14-3-3 proteins in lesion tissues and evaluate lesion activity. Separate the lymphocytes from the liver, spleen and abdominal cavity of mice, and use flow cytometry to detect and analyze the proportional changes of lymphocyte subsets in the liver, spleen and abdominal cavity of mice in each group. Results HE staining results showed that compared with the control group, significant inflammatory cell infiltration was observed in the lesion tissues of mice in the Az group and the Az+ABZ group. Immunohistochemical results indicated that compared with the control group, the expression levels of Em 14-3-3 and Em 18 proteins in the lesion tissues of mice in the Az group and the Az+ABZ group were significantly reduced(P<0.001). Flow cytometry analysis revealed that compared with the control group, the proportions of NK cells and B cells in the liver of mice in the Az group and the Az+ABZ group were significantly increased(P<0.01). In contrast, the proportion of T cells in the liver of mice in the Az+ABZ group was significantly decreased compared with the control group(P<0.05). Additionally, the proportion of B cells in the liver of mice in the Az group was significantly increased(P<0.01). A trend toward increased NK cell proportions and decreased T cell proportions was observed in the spleen of mice in the Az group and the Az+ABZ group compared with the control group(P>0.05). The proportion of B cells in the spleen of mice in the Az group was significantly increased(P<0.05). In the peritoneal cavity, the proportions of NK cells and B cells showed a trend toward increase, while T cells and CD8+ T cells showed a trend toward decrease in the Az group and the Az+ABZ group compared with the control group(P>0.05). Compared with Az group alone, the NK cell proportions in the liver, spleen and peritoneal lymphocytes showed an upward trend in the Az+ABZ group. Conclusion The intervention of heat shock protein 70 inhibitor Az combined with ABZ in a mouse model of E.m infection can inhibit lesion activity, promote the recruitment of NK cells to the liver, andmediate the inhibition of multilocular echinococcosis lesions.
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基本信息:
中图分类号:R532.32
引用信息:
[1]阿迪莱·多力坤,李寅时,邓冰清,等.热休克蛋白70抑制剂联合阿苯达唑干预对多房棘球蚴感染小鼠病灶活性和淋巴细胞亚群的影响[J].新疆医科大学学报,2025,48(03):277-285.
基金信息:
新疆维吾尔自治区“天山英才”培养计划—青年拔尖人才项目(2022TSYCCX0106); 省部共建中亚高发病成因与防治国家重点实验室开放项目(SKL-HIDCA-2022-1)
2025-03-15
2025-03-15