瘢痕灸预处理对原发性肝癌大鼠Wnt信号通路相关基因表达的影响
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摘要
目的:探讨瘢痕灸预处理肝俞及足三里穴对原发性肝癌大鼠Wnt信号传导通路相关基因表达的影响。方法:健康雄性Wistar大鼠48只随机分为空白组、模型组、预处理肝俞组和预处理足三里组,每组12只。预处理肝俞组和预处理足三里组均提前15 d施行瘢痕灸后,与模型组一同采用腹腔注射二乙基亚硝胺溶液(50mg/kg)建立肝癌模型。采用ELISA法检测血清中甲胎蛋白(alpha fetoprotein,AFP)、γ-谷胺酰转肽酶(Glutamyl transpeptidase,γ-GT)和岩藻糖苷酶(fucosidase,AFU)的表达量,RT-PCR法检测Wnt信号传导通路中Wnt3amRNA、Wnt10bmRNA的表达量,并观察大鼠肝脏组织形态学变化。结果:与模型组比较,预处理肝俞组、预处理足三里组的大鼠AFP、AFU、γ-GT和Wnt3amRNA、Wnt10bmRNA的表达量均降低,差异有统计学意义(P<0.05);与预处理肝俞组比较,预处理足三里组各指标表达量均显著升高,差异有统计学意义(P<0.05)。HE染色表明模型组肿瘤病变灶已形成,镜下可观察癌变细胞已深染,形态不规则,细胞核异常,增生结节较多;预处理足三里和预处理肝俞组两组癌变细胞数量较少,可见较少的结节形成。结论:瘢痕灸预处理肝俞和足三里穴均可以降低原发性肝癌大鼠血清中AFP、AFU、γ-GT和Wnt信号传导通路中Wnt3amRNA、Wnt10bmRNA的表达量,且瘢痕灸肝俞穴的预防作用更为显著。
Objective: To discuss the relative genetic expression influence on primary hepatic carcinoma rats' Wnt signal channel pre-processed on Ganshu( BL18) and Zusanli( ST36) acupoints by scarred moxibustion. Methods: Forty-eight healthy male Wistar rats were divided into the blank group,the model group,the pre-processed Ganshu( BL18)group,the pre-processed Zusanli( ST36) group with 12 in each group. The rats in pre-processed Zusanli( ST36) group and pre-processed Ganshu( BL18) group were injected diethylnitrosamine solution( 50 mg/kg) to build hepatic carcinoma model after they had been treated by scarred moxibustion. We used ELISA method to examine the expressions of alpha fetoprotein( AFP),Gl-utamyl transpeptidase( γ-GT),Fucosidase( AFU) and examined the expressions of Wnt3 amRNA and Wnt10 bmRNA in Wnt conductive signal channel with RT-PCR method and observed the transformation of histomorphology of rat ' s liver tissue. Results: Compared with the model group,the AFP,AFU,γ-GT and Wnt3 amRNA and Wnt10 bmRNA expressions in pre-processed Zusanli( ST36) and pre-processed Ganshu( BL18) groups decreased. The distinction had statistical meaning( P<0.05). Compared with the pre-processed Ganshu( BL18) group,the expression of each specification increased obviously and the distinction had statistical meaning( P<0.05). HE dye method proved thatthe focus on the model group was formed and carcinoma cell could be seen deep dyed under microscope which was abnormal form and nucleus,much more hyperplastic nodule. The pre-processed Ganshu( BL18) group and Zusanli( ST36)group had less carcinoma cell and hyperplastic nodule. Conclusion: The expressions of AFP,AFU and γ-GT in primary hepatic carcinoma rats' serum and Wnt3 amRNA and Wnt10 bmRNA in Wnt conductive signal channel could be decreased while scarred moxibustion on Ganshu( BL18) acupoint has more efficacy in precaution.
Objective: To discuss the relative genetic expression influence on primary hepatic carcinoma rats' Wnt signal channel pre-processed on Ganshu( BL18) and Zusanli( ST36) acupoints by scarred moxibustion. Methods: Forty-eight healthy male Wistar rats were divided into the blank group,the model group,the pre-processed Ganshu( BL18)group,the pre-processed Zusanli( ST36) group with 12 in each group. The rats in pre-processed Zusanli( ST36) group and pre-processed Ganshu( BL18) group were injected diethylnitrosamine solution( 50 mg/kg) to build hepatic carcinoma model after they had been treated by scarred moxibustion. We used ELISA method to examine the expressions of alpha fetoprotein( AFP),Gl-utamyl transpeptidase( γ-GT),Fucosidase( AFU) and examined the expressions of Wnt3 amRNA and Wnt10 bmRNA in Wnt conductive signal channel with RT-PCR method and observed the transformation of histomorphology of rat ' s liver tissue. Results: Compared with the model group,the AFP,AFU,γ-GT and Wnt3 amRNA and Wnt10 bmRNA expressions in pre-processed Zusanli( ST36) and pre-processed Ganshu( BL18) groups decreased. The distinction had statistical meaning( P<0.05). Compared with the pre-processed Ganshu( BL18) group,the expression of each specification increased obviously and the distinction had statistical meaning( P<0.05). HE dye method proved thatthe focus on the model group was formed and carcinoma cell could be seen deep dyed under microscope which was abnormal form and nucleus,much more hyperplastic nodule. The pre-processed Ganshu( BL18) group and Zusanli( ST36)group had less carcinoma cell and hyperplastic nodule. Conclusion: The expressions of AFP,AFU and γ-GT in primary hepatic carcinoma rats' serum and Wnt3 amRNA and Wnt10 bmRNA in Wnt conductive signal channel could be decreased while scarred moxibustion on Ganshu( BL18) acupoint has more efficacy in precaution.
引文
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[13]宁珠,殷芳,刘海,等.血清AFP、CEA、AFU、GGT-Ⅱ联合检测诊断原发性肝癌的临床价值[J].中国医药导报,2013,10:73-75.
[14]董飞波,高国生,徐晓珍,等.血清GP73和AFP等十项肿瘤标志物联合诊断HBs Ag阳性肝癌的研究[J].浙江医学,2014,36(2):166-159.
[15]Waisberg J,Saba GT.Wnt-/-beta-catenin pathway signaling in human hepatocellular carcinoma[J].World J Hepatol,2015,7(26):2631-2635.
[16]Chen L,Li M,Li Q,et al.DKK1 promotes hepatocellular carcinoma cell migration and invasion through beta-catenin/MMP7signaling pathway[J].Mol Cancer,2013(12):157.
[17]Tian J,He H,Lei G.Wnt/beta-catenin pathway in bone cancers[J].Tumour Biol,2014,35(10):9439-9445.
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[19]李圣杰,杨之斌.Wnt/β-catenin信号通路相关分子在肝癌靶向治疗中的研究进展[J].山东医药,2016,56(10):99-101.
[20]李伟,谢肖俊,陈胜林,等.Wnt/β-Catenin信号通路在肝癌发生中的分子机制研究[J].中国现代手术杂志,2014,18(1):3-8.
[21]赵玉洁,胡建鹏,王建敏,等.胃癌发生发展中β-连环蛋白异常表达与Wnt传导通路激活的关系[J].中国全科医学,2009,12(14):1365-1460.
[22]Hakim SG,Kosmehl H,Sieg P,et al.Altered expression of cell-cell adhesion moleculesβ-catenin/E-cadherin and related Wnt-signaling pathway in sporadic and syndromal keratocystic odontogenic umors[J].Clin Oral Investig,2011,15(3):321-328.
[23]Yuzgullu H,Benhaj K,Ozturk N,et al.Canonical Wnt signaling is an tagonized by noncanoncal Wnt5a in hepatocellular carcinoma cells[J].Mol Cancer,2009,8:90.
[24]柳杨,侯中伟,卢俊,等.麦粒灸“肝俞”对原发性肝癌癌前病变模型大鼠肝功能的影响[J].中国针灸,2015,7(35):702-706.
[2]贾文睿,侯中伟,睢明河.灸疗法防治原发性肝癌的研究进展[J].针灸临床杂志,2014,30(2):71-74.
[3]翁瑜君.浅谈瘢痕灸[J].河南中医,2009,3(29):289-290.
[4]李忠仁,方剑乔.实验针灸学[M].北京:中国中医药出版社,2007:244.
[5]周陈杰,宫绪萌,蔡理全,等.改良的从肝硬化到产生肝癌的大鼠模型[J].南方医科大学学报,2014,34(4):497-499.
[6]陈建军,陈建国,高晓阳,等.应用ROC曲线评价AFP、AFU、5'-NT在原发性肝癌诊断中的意义[J].实用预防医学,2012,(19):1402-1404.
[7]张莉莉,王立.肝癌癌前病变防治机制的研究概况[J].中国中西医结合杂志,2010,30(9):1003-1008.
[8]Fattovich G,Stroffolini T,Zagni I,et al.Hepatocellular Carcinoma in Cirrhosis:Incidence and Risk Factors[J].Gsatroenterology,2004,127:s35-s50.
[9]Polakis P.The many ways of Wnt in cancer[J].Curr Opin Genet Dev,2007,17(1):45-51.
[10]张天明,王玉平,周永宁.肝癌肿瘤标记物的研究进展[J].胃肠病学和肝病学杂志,2016,25(8):852-855.
[11]Mogensen CE.Twelve shifting paradigms in diabetic renal disease and hypertension[J].Diabetes Res Clin Pract,2008,11(13):82.
[12]羊文芳,郑文雯,苏显都,等.肿瘤标记物在肝癌患者血清中的表达及其诊断价值[J].解放军医学院学报,2016,37(9):976-1000.
[13]宁珠,殷芳,刘海,等.血清AFP、CEA、AFU、GGT-Ⅱ联合检测诊断原发性肝癌的临床价值[J].中国医药导报,2013,10:73-75.
[14]董飞波,高国生,徐晓珍,等.血清GP73和AFP等十项肿瘤标志物联合诊断HBs Ag阳性肝癌的研究[J].浙江医学,2014,36(2):166-159.
[15]Waisberg J,Saba GT.Wnt-/-beta-catenin pathway signaling in human hepatocellular carcinoma[J].World J Hepatol,2015,7(26):2631-2635.
[16]Chen L,Li M,Li Q,et al.DKK1 promotes hepatocellular carcinoma cell migration and invasion through beta-catenin/MMP7signaling pathway[J].Mol Cancer,2013(12):157.
[17]Tian J,He H,Lei G.Wnt/beta-catenin pathway in bone cancers[J].Tumour Biol,2014,35(10):9439-9445.
[18]何益.Wnt信号通路与肝纤维化[J].国际鲜花病杂志,2008,28(1):57-59.
[19]李圣杰,杨之斌.Wnt/β-catenin信号通路相关分子在肝癌靶向治疗中的研究进展[J].山东医药,2016,56(10):99-101.
[20]李伟,谢肖俊,陈胜林,等.Wnt/β-Catenin信号通路在肝癌发生中的分子机制研究[J].中国现代手术杂志,2014,18(1):3-8.
[21]赵玉洁,胡建鹏,王建敏,等.胃癌发生发展中β-连环蛋白异常表达与Wnt传导通路激活的关系[J].中国全科医学,2009,12(14):1365-1460.
[22]Hakim SG,Kosmehl H,Sieg P,et al.Altered expression of cell-cell adhesion moleculesβ-catenin/E-cadherin and related Wnt-signaling pathway in sporadic and syndromal keratocystic odontogenic umors[J].Clin Oral Investig,2011,15(3):321-328.
[23]Yuzgullu H,Benhaj K,Ozturk N,et al.Canonical Wnt signaling is an tagonized by noncanoncal Wnt5a in hepatocellular carcinoma cells[J].Mol Cancer,2009,8:90.
[24]柳杨,侯中伟,卢俊,等.麦粒灸“肝俞”对原发性肝癌癌前病变模型大鼠肝功能的影响[J].中国针灸,2015,7(35):702-706.