目的 探究氯胺酮联合吉西他滨对脑胶质瘤大鼠免疫逃逸、瘤体体积及MKK3/6信号通路的影响。 方法 选取50只SPF级SD雄性大鼠，随机分为正常（NO）组，模型（MO）组，氯胺酮（KM）组，吉西他滨（GT）组，氯胺酮联合吉西他滨（KG）组，每组10只，对MO、KM、GT、KG采用立体定位法进行脑胶质瘤建模，NO组不建立该模型。建模成功后，对KM组腹腔注射100 mg/kg氯胺酮，GT组腹腔注射150 mg /kg吉西他滨，KG组腹腔注射100 mg/kg氯胺酮及150 mg /kg吉西他滨，NO组、MO组同期腹腔注射同体积生理盐水，HE染色检测脑胶质瘤组织病理形态，ELISA及RT-PCR检测免疫逃逸相关因子，免疫印迹法检测MKK3/6蛋白表达。 结果 与NO组相比，MO组IL-4、IL-10、TGF-β1含量及mRNA表达升高（P<0.05），MKK3/6蛋白表达明显降低（P<0.05）；与MO组相比，KM组、GT组、KG组IL-4、IL-10、TGF-β1含量及mRNA表达降低（P<0.05），肿瘤抑制率、MKK3/6蛋白表达升高（P<0.05），且GT、KM组比较无明显差异（P>0.05），KG组比GT组变化明显（P<0.05）；NO组脑组织结构正常，细胞排列整齐，MO组脑组织遭到破坏，可见大量浸润成长的肿瘤细胞及淋巴细胞浸润、癌细胞核染色变深；与MO组比较，KM、GT、KG组病理结构明显改善，癌细胞核染变浅。 结论 氯胺酮联合吉西他滨对脑胶质瘤大鼠具有显著疗效，可有效抑制免疫逃逸，并对MKK3/6表达具有显著的调控作用。
Abstract
Objective To investigate the effects of ketamine combined with gemcitabine on immune escape, tumor volume and MKK3/6 signaling pathway in brain glioma of rats. Methods Fifty SPF SD male rats were randomly divided into normal (NO), model (MO), ketamine (KM), gemcitabine (GT), ketamine combined with gemcitabine (KG) groups, with 10 in each group. Stereotaxic method was used to establish brain glioma modeling in MO, KM, GT and KG groups, while the model was not established in NO group. After establishing the modeling, the KM group was intraperitoneally injected with 100 mg/kg ketamine, the GT was intraperitoneally injected with 150 mg/kg gemcitabine, the KG with 100 mg/kg ketamine and 150 mg/kg gemcitabine, and NO and MO with the same volume of normal saline at the same time. HE staining was used to detect histopathologic morphology, ELISA and RT-PCR were used to detect immune escape related factors, and MKK3/6 protein expression was detected by Western blot. Results Compared with NO, the contents and mRNA expression of IL-4, IL-10 and TGF-β1 in MO groups were markedly increased (P<0.05), while the expression of MKK3/6 protein was significantly decreased (P<0.05). Compared with MO, the contents and mRNA expression of IL-4, IL-10 and TGF-β1 in KM, GT and KG groups were obvious decreased (P<0.05), while the tumor inhibition rate and MKK3/6 protein level were increased (P<0.05), however, which had no significant differences between GT and KM groups (P>0.05). The changes in KG were more significant than those in GT group (P<0.05). In NO, the brain tissue structure was normal and the cells were arranged neatly, while in group MO, the brain tissue was destroyed, and a large number of infiltrating tumor cells and lymphocytes could be seen, and the nuclear staining of cancer cells became deeper. Compared with group MO, the pathological structure of KM, GT and KG groups was significantly improved, and the nuclear staining of cancer cells became lighter. Conclusions Ketamine combined with gemcitabine has a significant effect on brain glioma, which can effectively inhibit immune escape, and regulate MKK3/6 expression.
关键词
氯胺酮； /
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吉西他滨； /
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脑胶质瘤； /
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免疫逃逸； /
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瘤体体积； /
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MKK3/6信号通路
Key words
Ketamine; /
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Gemcitabine; /
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Brain glioma; /
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Immune escape; /
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Tumor volume; /
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MKK3/6 signaling
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基金
国家自然科学基金资助项目（81860450）
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