郭丽新,钟文,姜玲
GUO Lixin,ZHONG Wen,JIANG Ling

• 1:黑龙江中医药大学

摘要(Abstract)：

目的:蛹虫草多糖(CMPS)对D-半乳糖所致衰老模型小鼠肝组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)水平及移植型小鼠腹水型肝癌(H22)瘤组织B-淋巴细胞瘤-2(Bcl-2)、B-淋巴细胞瘤-2相关X(Bax) mRNA表达的影响。方法:75只小鼠随机均分为对照组、模型组及CMPS高、中、低剂量组;模型组及3个CMPS组小鼠每日给予0. 3 g/kg D-半乳糖溶液颈部皮下注射建立衰老模型,同时模型组每日给予0. 033 g/kg肌肉注射生理盐水,CMPS高、中及低剂量组分别给予100、50及25 mg/kg CMPS大腿外侧肌肉注射,对照组分别在颈部及大腿外侧肌肉注射等量生理盐水,每日注射前记录小鼠行动、进食、饮水、生长、发育、毛色等情况;连续注射6周后检测小鼠肝脏组织SOD、CAT活性及MDA含量;另取30只腋下移植瘤小鼠,均分为模型组、CMPS中、低剂量组,CMPS中、低剂量组小鼠分别注射50、25 mg/kg CMPS(1次/d),模型组给予等量生理盐水处理,连续10 d;第10天时,取瘤组织采用RT-PCR方法检测H22瘤组织中Bcl-2及Bax mRNA表达水平。结果:观察可见,衰老模型小鼠活动减少,精神萎靡,倦怠嗜睡,行动迟缓,形体瘦弱,体力较弱,皮毛松散,毛色枯槁缺乏光泽,皮肤弹性差,进食明显减少等衰老体征; CMPS组小鼠活动量较模型组增加,活力更强,生长发育状况与精神状况较好,皮毛滑顺有光泽;注射6周后,模型组小鼠肝脏组织SOD、CAT活性较对照组显著下降,MDA含量较对照组显著升高(P <0. 05); 3个CMPS剂量组小鼠肝组织SOD、CAT活性较模型组升高,MDA含量较模型组降低(P <0. 05),与模型组比较,CMPS两个剂量组H22瘤组织Bcl-2 mRNA表达水平显著降低,BaxmRNA的表达水平显著升高(P <0. 05);与CMPS低剂量组比较,CMPS中剂量组变化更明显(P <0. 05)。结论:CMPS可有效减轻D-半乳糖所致衰老小鼠的肝组织损伤,其机制可能与CMPS抗氧化损伤,提高氧自由基清除能力有关; CMPS可能通过氧化应激抑制及调控凋亡相关蛋白Bcl-2、Bax的mRNA表达水平参与H22荷瘤小鼠肿瘤细胞凋亡过程。
Objective: To investigate the effects of Cordyceps militaris Polysaccharide( CMPS) on the level of superoxide dismutase( SOD),catalase( CAT) and malondialdehyde( MDA) in liver tissue of aging rats induced by D-galactose and on the expression of B-Lymphoma 2( Bcl-2) and B lymphocytoma-2 related protein X( Bax) mRNA of ascites hepatocellular carcinoma( H22) in transplanted mice.Methods: 75 rats were randomly divided into control group,model group and CMPS high,medium and low dose groups. The aging model was established by subcutaneous injection of 0. 3 g/kg D-galactose solution into the neck of model group and 3 CMPS groups were intramuscularly injected with normal saline for 0. 033 g/kg daily. The high,medium and low dose groups of CMPS were given100 mg/kg,50 mg/kg and 25 mg/kg CMPS intramuscular injection of the lateral thigh,respectively.The control group was injected with the same amount of saline into the neck and the lateral thigh. The activity,feeding,drinking,growth,development and coat color of rats were recorded before the daily injection,and SOD,CAT activity and MDA content in liver tissue of rats were detected after 6-week continuous injection. Another 30 subaxillary transplanted tumor rats were divided into three groups:model group,CMPS medium dose group and low dose group. In CMPS,the rats of medium and low dose group were injected with 50 mg/kg,25 mg/kg CMPS( once a day) respectively,and the model group was treated with the same amount of normal saline for 10 days. On the 10 th day,the expression level of Bcl-2 and Bax mRNA in H22 tumor tissue was detected by RT-PCR method. Results: It can be seen that the aging model rats had decreased activity,weak spirit,lethargy,slow movement,thin body,weak physical strength,loose fur,haggard and lack of luster,poor elasticity of skin,and obvious decrease in food intake. On the contrast,the activity of CMPS group was higher than that of model group; the CMPS group showed stronger activity,better growth and development and smoother and glossier fur. After 6 weeks of injection,the activity of SOD,CAT in the liver tissue of the model group decreased significantly while the content of MDA increased significantly,compared with the control group( P < 0. 05). The activity of SOD,CAT in liver tissue of three CMPS groups increased while the content of MDA decreased,compared with the model group( P < 0. 05). Compared with the model group,the expression of Bcl-2 mRNA in H22 tumor tissues in two dose groups of CMPS decreased significantly( P < 0. 05) and the expression of Bax mRNA increased significantly( P < 0. 05). Compared with the low dose CMPS group,the CMPS medium dose group changed more obviously( P < 0. 05).Conclusion: CMPS can effectively reduce the liver damage caused by D-galactose-induced aging rats,and the mechanism may be related to CMPS anti-oxidative damage,improvement of the ability of scavenging oxygen free radicals. CMPS may participate in the apoptotic process of H22 tumor-bearing rats by inhibiting oxidative stress and regulating the expression of Bcl-2 and Bax.

关键词(KeyWords)： 蛹虫草多糖;D-半乳糖;超氧化物歧化酶;过氧化氢酶;丙二醛;凋亡
Cordyceps militaris polysaccharide;D-galactose;superoxide dismutase;catalase;malondialdehyde;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年项目(81503464);; 黑龙江省留学归国人员科学基金(LC2015032);; 黑龙江省普通本科高等学校青年创新人才培养计划基金(UNPYSCT-2016079);; 黑龙江中医药大学博士创新基金(2013bs01)

作者(Author): 郭丽新,钟文,姜玲
GUO Lixin,ZHONG Wen,JIANG Ling

DOI: 10.19367/j.cnki.1000-2707.2019.02.010

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