针灸中药对模型小鼠肿瘤细胞bcl-2,bax和PCNA表达的影响
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摘要
目的：本研究在前期临床及实验研究的基础上，从肿瘤细胞凋亡相关基因和增殖细胞核抗原角度研究针刺、艾灸和中药促进肿瘤细胞凋亡、抑制肿瘤细胞增殖的机制，为进一步扩展这几种疗法的临床应用提供科学实验依据。
方法：按体重将120只小鼠随机分为中药低剂量组、中药高剂量组、针刺组、艾灸组、模型组和空白组。除空白组外，其余各组采用腋下注射法建立小鼠S180模型。针刺组针刺足三里并在接种部位围刺三针。中药高、低剂量组采用导师自拟方药(浓度分别为每ml相当于3.4g和1.7g中药生药)。艾灸组采用自制0.6mm×10cm小艾条，各组每日处理一次，模型组和空白组进行与前五组相同程度的捉抓。连续处理10天后，脱颈处理，取肿瘤组织(空白组除外)，计算抑瘤率。采用免疫组织化学技术检测肿瘤组织的bcl-2、bax蛋白和PCNA的表达。采用计算机图像分析技术对其进行定量分析，所得数据用SPSS11.0软件进行单因素方差分析(One-Way ANOVA)。
结果：
1．各组抑瘤率分别为：中药低剂量组30.5％，中药高剂量组47.9％，，针刺组37.7％，艾灸组34％；
2．各组bcl-2蛋白表达显著低于模型组，bax蛋白表达显著高于模型组，PCNA表达显著低于模型组(P＜0.01)。
结论：
针刺、艾灸和中药均能显著降低S180模型小鼠肿瘤细胞bcl-2蛋白、PCNA表达，提高bax蛋白表达，降低bcl-2／bax比值，这是其发挥抗肿瘤作用的机制之一。
Objective:Based on the previous clinical and experimental studies, this research deals with the mechanism that acupuncture ,moxibustion and Chinese herbs promote apoptosis and inhibit proliferation in tumor cell via genes relative to supply new experimental proofs to cancer-treating with these TCM therapies.
Method:Excluding the control group ,the rest were injected single-cell suspension at the left axilla to set up SI80 model. The 120 mice were randomly divided into low-dosage group,high-dosage group,acupuncture group group,model group and control group according to their body weight. The low-dosage group and high dosage group were treated with self-made herb solution at a dosage of 3.4 ml and 1.7g/ml respectively.The moxibustion group was treated with self-made moxa stick (0.6mmX 10cm) at RN4.The acupuncture group was needled in ST36 point and round the injecting cite. These 5 groups were treated 1 time per day for 10 days successively.The model group and control group were stimulated by grasp at the same time and with the same strength as those 5 groups.At the list day. all groups were sacrificed, and the tumor tissue was taken out .Then the inhibiting-tumor rates were calculated,and the expression rates of bcl-2, bax protein and PCNA were determined by immunohistochemical technique with computerized image processing The data were analyzed with one-way ANOVA by means of SPSS 11.0.
Result:
1.The inhibiting-tumor rates of these 5 groups were low-dosage group 30.5%, high dosage group 49.7%, acupuncture group 37.7%, moxibustion group 34%.
2. The expression rates of bcl-2 protein and PCNA in tumor cell in all TCM therapies groups was significantly lower and the expression rates of bax protein were significantly higher than those in model group(p<0.01).
Conclusion:
Acupuncture ,moxibustion and Chinese herbs can suppress the expression of bcl-2 and PCNA ,enhance the expression of bax and lower the ratio of bcl-2/bax in tumor cell of s180 model mice,which is one of mechanisms for these TCM therapies to exert their antitumor efficacy.
方法：按体重将120只小鼠随机分为中药低剂量组、中药高剂量组、针刺组、艾灸组、模型组和空白组。除空白组外，其余各组采用腋下注射法建立小鼠S180模型。针刺组针刺足三里并在接种部位围刺三针。中药高、低剂量组采用导师自拟方药(浓度分别为每ml相当于3.4g和1.7g中药生药)。艾灸组采用自制0.6mm×10cm小艾条，各组每日处理一次，模型组和空白组进行与前五组相同程度的捉抓。连续处理10天后，脱颈处理，取肿瘤组织(空白组除外)，计算抑瘤率。采用免疫组织化学技术检测肿瘤组织的bcl-2、bax蛋白和PCNA的表达。采用计算机图像分析技术对其进行定量分析，所得数据用SPSS11.0软件进行单因素方差分析(One-Way ANOVA)。
结果：
1．各组抑瘤率分别为：中药低剂量组30.5％，中药高剂量组47.9％，，针刺组37.7％，艾灸组34％；
2．各组bcl-2蛋白表达显著低于模型组，bax蛋白表达显著高于模型组，PCNA表达显著低于模型组(P＜0.01)。
结论：
针刺、艾灸和中药均能显著降低S180模型小鼠肿瘤细胞bcl-2蛋白、PCNA表达，提高bax蛋白表达，降低bcl-2／bax比值，这是其发挥抗肿瘤作用的机制之一。
Objective:Based on the previous clinical and experimental studies, this research deals with the mechanism that acupuncture ,moxibustion and Chinese herbs promote apoptosis and inhibit proliferation in tumor cell via genes relative to supply new experimental proofs to cancer-treating with these TCM therapies.
Method:Excluding the control group ,the rest were injected single-cell suspension at the left axilla to set up SI80 model. The 120 mice were randomly divided into low-dosage group,high-dosage group,acupuncture group group,model group and control group according to their body weight. The low-dosage group and high dosage group were treated with self-made herb solution at a dosage of 3.4 ml and 1.7g/ml respectively.The moxibustion group was treated with self-made moxa stick (0.6mmX 10cm) at RN4.The acupuncture group was needled in ST36 point and round the injecting cite. These 5 groups were treated 1 time per day for 10 days successively.The model group and control group were stimulated by grasp at the same time and with the same strength as those 5 groups.At the list day. all groups were sacrificed, and the tumor tissue was taken out .Then the inhibiting-tumor rates were calculated,and the expression rates of bcl-2, bax protein and PCNA were determined by immunohistochemical technique with computerized image processing The data were analyzed with one-way ANOVA by means of SPSS 11.0.
Result:
1.The inhibiting-tumor rates of these 5 groups were low-dosage group 30.5%, high dosage group 49.7%, acupuncture group 37.7%, moxibustion group 34%.
2. The expression rates of bcl-2 protein and PCNA in tumor cell in all TCM therapies groups was significantly lower and the expression rates of bax protein were significantly higher than those in model group(p<0.01).
Conclusion:
Acupuncture ,moxibustion and Chinese herbs can suppress the expression of bcl-2 and PCNA ,enhance the expression of bax and lower the ratio of bcl-2/bax in tumor cell of s180 model mice,which is one of mechanisms for these TCM therapies to exert their antitumor efficacy.
引文
1 曹世龙．肿瘤学——新理论与新技术·世界和中国肿瘤发病和死亡情况．上海科技教育出版社，1997年第一版：509～519
2 李连弟，鲁凤珠，张思维，等．中国恶性肿瘤死亡率20年变化趋势和近期预测分析．中华肿瘤杂志，1997：19：3
3 杨柱，陈学习．肿瘤的中医病因病机初探．辽宁中医杂志，2002；29：197
4 王永炎主编．中医内科学．上海科学技术出版社，1997，91
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9 李艳春．针刺对晚期肿瘤患者的止痛研究．长春中医学院学报，1998：14(3)：29
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13 李晔．针刺治疗恶性肿瘤放化疗副反应的临床研究．中国针灸，1997；17(6)：327
14 路玫．穴位敷贴治疗癌症化疗患者白细胞减少的临床观察．针刺研究，1998：23(2)：115
15 马泽云．太乙雷火神针治疗白细胞减少症的临床研究．中国针灸，1997；17(4)：207
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17 黄喜梅，等．针刺治疗癌症化疗中胃肠反应的疗效观察．中国中西医结合杂志，1994；14(10)：618
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2 李连弟，鲁凤珠，张思维，等．中国恶性肿瘤死亡率20年变化趋势和近期预测分析．中华肿瘤杂志，1997：19：3
3 杨柱，陈学习．肿瘤的中医病因病机初探．辽宁中医杂志，2002；29：197
4 王永炎主编．中医内科学．上海科学技术出版社，1997，91
5 杨运宽，等．针灸防治肝癌概况．中国针灸，2002：22(9)：638
6 曾金雄，戴西湖．毫米波循经传导治疗癌症疼痛患者51例临床观察．中西医结合肿瘤学杂志，1996，(1)：43
7 徐淑英，等．针刺足三里穴治疗癌性腹痛69例临床观察．针刺研究，1991；16(3-4)：244
8 左秀玲．孔最穴在肺癌止痛中的作用．河北中医，1991：13(3)：4
9 李艳春．针刺对晚期肿瘤患者的止痛研究．长春中医学院学报，1998：14(3)：29
10 闫继勒，周长运，等．重组人白细胞介素-2穴位注射对癌症发热的疗效观察．中国针灸，1999；(7)：415
11 蔡庆春，刘建平，原春荣．针灸升白细胞的机制探讨．河南中医药学刊，1998：(5)：17
12 赵喜新，王和平，田开宇，等．针灸对化疗小鼠骨髓粒系细胞动态变化的影响．中国针灸，2000；(3)：172
13 李晔．针刺治疗恶性肿瘤放化疗副反应的临床研究．中国针灸，1997；17(6)：327
14 路玫．穴位敷贴治疗癌症化疗患者白细胞减少的临床观察．针刺研究，1998：23(2)：115
15 马泽云．太乙雷火神针治疗白细胞减少症的临床研究．中国针灸，1997；17(4)：207
16 杨金洪．放化疗对恶性肿瘤患者内分泌功能的影响及针刺调节作用．针刺研究，1995：(1)：1
17 黄喜梅，等．针刺治疗癌症化疗中胃肠反应的疗效观察．中国中西医结合杂志，1994；14(10)：618
18 常瑛．654-Ⅱ、异丙嗪双足三里穴位注射对抗化疗呕吐的临床观察．北京中医，1991；(4)：56
19 郁美娟，等．针刺治疗放化疗所致造血功能损害的临床观察．中国针灸，1994：14(5)：4
20 欧阳群，等．隔盐壮灸神阙穴对机体免疫功能的影响：动物实验及临床验证．新中医，1992；24(2)30，32
21 吴耀持．毫米波循经传导穴位辐照治疗肿瘤的作用．上海针灸杂志，1998；17(2)：15
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