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蛞蝓提取物对环磷酰胺治疗肝癌的增效减毒作用
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| 篇名： | 蛞蝓提取物对环磷酰胺治疗肝癌的增效减毒作用 |
| TITLE： | Effect of Increasing Efficacy and Decreasing Toxicity of Limax Extract on Cyclophosphamide in the Treatment of Hepatocellular Carcinoma |
| 摘要： | 目的：探讨蛞蝓提取物（LE）对环磷酰胺（CTX）治疗肝癌的增效减毒作用。方法：将小鼠随机分为正常组，模型组，CTX组（0.02g/kg），LE低、中、高剂量组（即LEL、LEM、LEH组，0.6、1.2、2.4g/kg），CTX+LE低、中、高剂量联用组（即CTX+LEL、CTX+LEM、CTX+LEH组，剂量同各单药组），每组10只。除正常组外，其余各组小鼠均于左腋下接种小鼠肝癌细胞H22以复制荷瘤模型。接种24h后，正常组和模型组小鼠灌胃生理盐水，各给药组小鼠分别灌胃相应药物，每日1次，连续10d。末次给药后第2天，观察各组小鼠一般情况，测定体质量和胸腺指数（LI）、脾指数（SI），检测抑瘤率，并采用金氏公式评估联合用药的效果（q）；检测模型组、CTX组和各联用组小鼠的白细胞（WBC）、红细胞（RBC）、血小板（PLT）计数以及血清中丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（ALT）、血肌酐（Scr）、尿素氮（BUN）含量，并采用比色法和酶联免疫吸附测定（ELISA）法分别检测上述各组小鼠肿瘤组织中丙二醛（MDA）含量和超氧化物歧化酶（SOD）、还原型谷胱甘肽（GSH）活性以及血管内皮生长因子（VEGF）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）水平；采用免疫组化法检测模型组、CTX组和CTX+LEM组小鼠肿瘤组织中癌基因蛋白（p53、Bcl-2、Bax）的表达水平。结果：模型组小鼠精神不佳，且出现多饮、多食症状；其体质量、TI、SI虽较正常组无明显异常（P＞0.05），但其WBC计数、AST含量均显著升高，ALT、BUN含量均显著降低（P＜0.05或P＜0.01）。与模型组比较，各给药组小鼠上述症状有所改善；各给药组的瘤体质量，CTX组的TI、SI以及各联用组的TI均显著降低，但LEL组、LEH组的瘤体质量，LE单用组及各联用组的TI、SI均显著高于CTX组，各联用组的瘤体质量均显著低于CTX组（P＜0.05或P＜0.01）；各给药组的抑瘤率为29.58％～72.08％，CTX+LEL组、CTX+LEM组、CTX+LEH组的q值分别为1.03、0.97、0.86。与模型组比较，CTX组小鼠的WBC计数和AST、BUN含量，各联用组的MDA含量，各给药组的VEGF、TNF-α、IL-6水平以及CTX组和CTX+LEM组的Bcl-2表达水平均显著降低，各给药组的SOD、GSH活性以及CTX+LEM组的p53表达水平，CTX组和CTX+LEM组的Bax表达水平均显著升高（P＜0.05或P＜0.01）；但各联用药组的WBC计数和AST含量以及CTX+LEM组的ALT含量，CTX+LEH组的SOD活性和CTX+LEM组的GSH活性均显著高于CTX组，CTX+LEH组的MDA含量，CTX+LEM组、CTX+LEH组的VEGF、TNF-α水平和各联用组的IL-6水平均显著低于CTX组（P＜0.05或P＜0.01）。结论：LE和CTX联用具有相加的抑瘤作用，且LE可减少CTX致小鼠免疫功能减退、骨髓抑制等毒性反应，具增效减毒的作用。这种作用可能与抗氧化应激、抑制血管生成和炎症因子分泌以及调控凋亡蛋白的表达有关。 |
| ABSTRACT： | OBJECTIVE：To investigate the effect of increasing efficacy and decreasing toxicity of Limax extract （LE）on cyclophosphamide（CTX）in the treatment of hepatocellular carcinoma. METHODS ：The mice were randomly divided into normal group，model group ，CTX group （0.02 g/kg），LE low-dose ，medium-dose and high-dose groups （LEL，LEM，LEH group ，0.6，1.2，2.4 g/kg），CTX+LE low-dose ，medium-dose and high-dose combination groups （CTX+LEL，CTX+LEM，CTX+ LEH group ，the same dose as single drug group ），with 10 huangrenbin518@163.com mice in each group. Except for normal group ，other groups were inoculated with hepatoma cells H 22 in the left ar mpit to establish tumor bearing models. After 24 h of inoculation ，normal group and model group were intragastrically given normal saline ， and administration groups were intragastrically given corresponding drugs ，once a day ，for 10 days. On the second day after the last administration ，the general conditions of mice in each group were observed ；the body mass ，thymus index （LI），spleen index （SI）were measured ；the tumor inhibition rate was detected. The effect （q）of combination therapy was evaluated by King ’s formula . The counts of WBC ，RBC and PLT ，serum contents of ALT ，ALT，Scr and BUN were detected in model group ，CTX group and combination groups ，and the contents of MDA，SOD and GSH ，the levels of VEGF ，TNF-α and IL-6 in the tumor tissue were detected by colorimetry and ELISA in above groups. The protein expression of oncogenes （p53，Bcl-2 and Bax ）were detected by immunohistochemical method in model group，CTX group and CTX+LEM group. RESULTS ：The mice in the model group were in poor spirit and had symptoms of excessive drinking and eating ；although the body weight ，TI and SI were not significantly abnormal compared with normal group （P＞0.05），WBC count and AST content were significantly increased ，ALT and BUN contents were significantly decreased （P＜ 0.05 or P＜0.01）. Compared with model group ，above symptoms of mice were all improved in administration groups. The tumor weight of administration groups ，TI and SI of CTX group and TI of combination groups were decreased significantly ，but tumor weight of LEL group and LEH group ，TI and SI of LE single groups and combination groups were significantly higher than CTX group；tumor weight of combination groups were significantly lower than CTX group （P＜0.05 or P＜0.01）. The tumor inhibition rates of administration groups were 29.58％-72.08％. The q values of CTX+LEL group ，CTX+LEM group and CTX+LEH group were 1.03，0.97 and 0.86，respectively. Compared with model group ，WBC count ，AST and BUN contents of CTX group ，MDA contents of combination groups ，VEGF，TNF-α and IL-6 levels of administration groups ，the protein expression of Bcl- 2 in CTX group and CTX+LEM group were decreased significantly ；the activities of SOD and GSH of administration groups ，the protein expression of p 53 in CTX+LEM group and Bax in CTX group ，CTX+LEM group were increased significantly （P＜0.05 or P＜ 0.01）；WBC counts and AST contents of administration groups ，ALT content of CTX+LEM group ，SOD activity of CTX+LEH group and GSH activity of CTX+LEM group were all significantly higher than those of CTX group ；MDA content of CTX+LEH group，VEGF and TNF-α levels of CTX+LEM group and CTX+LEH group，IL-6 levels of administration groups were all significantly lower than CTX group （P＜0.05 or P＜0.01）. CONCLUSIONS ：LE combined with CTX can increase the anti-tumor effect，and LE can reduce the toxicity of CTX induced immunosuppression and bone marrow suppression in mice ，with effect of increasing efficacy and decreasing toxicity. The effect may be related to antioxidant stress ，inhibition of angiogenesis and secretion of inflammatory factors ，and regulation of apoptosis protein expression. |
| 期刊： | 2021年第32卷第01期 |
| 作者： | 黄天敏，杨映霞，张宏亮，陈俐秀，黄湘，谢金魁，黄仁彬，杨玉芳 |
| AUTHORS： | HUANG Tianmin， YANG Yingxia，ZHANG Hongliang，CHEN Lixiu，HUANG Xiang，XIE Jinkui，HUANG Renbin，YANG Yufang |
| 关键字： | 蛞蝓提取物；肝癌；抗氧化应激；血管生成；细胞凋亡；增效减毒；小鼠 |
| KEYWORDS： | Limax extract ；Hepatocellular carcinoma ；Antioxidant stress ；Angiogenesis；Cell apoptosis ；Increasing efficacy |
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