傣药牙帕汤抑制小鼠Lewis肺癌生长转移及小鼠免疫功能的影响

Experimental study of Dai medicine Yapa Tang on inhibiting the lewis lung carcinoma in mice growth and metastasis and effects on immune function

目的：探讨傣药牙帕汤对小鼠Lewis肺癌的抑制作用及其对小鼠免疫功能的影响。方法C57BL/6小鼠右腋皮下注射Lewis肺癌细胞（lewislungcarcinoma，LLC），采用随机双盲法（随机抽取仅留有一侧开口的密闭黑盒子中的30只雄性小鼠，放入事先备好的6个鼠笼中，雌性小鼠依此法进行分组），将接种的60只小鼠分为空白对照组、牙帕汤高、中、低剂量组、环磷酰胺组、牙帕汤与环磷酰胺联合组各10只。各组均从接种后第3天给药，空白对照组、牙帕汤高、中、低剂量组按0.4ml/（只·d）分别灌胃，给予0.9%生理盐水、高、中、低浓度的牙帕汤；环磷酰胺组每隔2d腹腔注射环磷酰胺0.2ml/只；牙帕汤与环磷酰胺联合组按0.4ml/（只·d）灌胃给予中剂量牙帕汤、每隔2d按0.2ml/只腹腔注射环磷酰胺。第21天，摘眼球取血，取肿瘤组织、脾、胸腺并称重，计算抑瘤率、脾指数和胸腺指数；采用酶联免疫吸附法检测血清中白细胞介素-2（IL-2）、白细胞介素-10（IL-10）含量。结果牙帕汤高剂量组、环磷酰胺组、牙帕汤与环磷酰胺联合组的瘤重分别为（3.46±0.39）g、（2.39±1.04）g、（2.30±0.76）g，均低于空白对照组（5.21±0.50）g，差异均有统计学意义（P＜0.05），抑瘤率分别为33.69%、54.12%、56.00%；牙帕汤低、中、高各剂量组的胸腺指数分别为（2.16±0.69）、（2.24±0.76）、（2.23±0.63），脾指数分别为（16.82±3.14）、（15.82±1.72）、（17.08±3.65），均高于空白对照组（1.94±0.68）、（15.17±3.53）；环磷酰胺组（1.72±0.58）、（14.82±3.78）下降较明显。与空白对照组比较，各给药组的肺转移灶数目均降低，但差异无统计学意义（P＞0.05）。空白对照组IL-2、IL-10分别为（883.54±181.49）ng/L、（1106.86±343.79）ng/L，牙帕汤低、中、高各剂量组的IL-2水平明显提高，分别为（1732.29±100.52）ng/L、（1813.33±168.32）ng/L、（2275.63±394.76）ng/L；IL-10分别为（834.02±271.97）ng/L、（636.83±270.56）ng/L、（682.08±147.85）ng/L水平显著降低。结论牙帕汤可以抑制Lewis肺癌细胞生长，降低肺转移灶数目，调节免疫细胞因子IL-2、IL-10的表达水平。

Objective To study the Dai medicine Yapa Tang inhibition on mice Lewis lung carcinoma growth and metastasis,and to explore the impact of the drug on mice immune function. Methods Subcutaneous injection LLC（Lewis lung carcinoma cells）in C57BL/6 mice’s right armpit was to establish rat models. 60 successful tumor-bearing mice were selected and thenafter inoculating LLC were randomly divided into 6 groups, namely the control group,Yapa Tang low-dose group,Yapa Tang mid-dose group, Yapa Tang high-dose group,cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group.Then,after inoculated 3 days,each group was administrated with medicines.The first four groups wereintragastricly administrated with 0.9% saline, low-dose, mid-dose and high-dose Yapa Tang with0.4ml respectively;the cyclophosphamide group was administrated with cyclophosphamide,and Yapa Tang mid-dose combining cyclophosphamide group was administrated with middle dose Yapa Tang and cyclophosphamide. After 21days, blood was taken from eyeballs. Tumor tissue, spleen, thymus were gotten and weighted to calculate inhibitory rate,spleen and thymus index, enzyme-linked immunosorbent assay （Elisa） was adopted to detect Interleukin-2（IL-2）, Interleukin-10（IL-10）and their contents. Results The tumor weight of Yapa Tang high-dose group, cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group was 3.46±0.39, 2.39±1.04 and 2.30±0.76 respectively,all lower than the control group, which was 5.21±0.50, showing significant difference（P〈0.05）, with the inhibition rate were33.69%, 54.12%, 56.00%. The thymus and spleen index of Yapa Tang low-dose group, middle-dose group and high-dose group was2.16±0.69, 2.24± 0.76, 2.23 ± 0.63, 16.82 ± 3.14, 15.82 ± 1.72, 17.08 ± 3.65, significantly higher than that of the control group,which was1.94±0.6, 15.17±3.53. The IL-2, IL-10 level of control group were（883.54±181.49）ng/L, （1 106.86±343.79）ng/L. Yapa Tanggroupsshowed an increase in IL-2（1 732.29±100.52）ng/L, （1 813.33± 168.32）ng/L, （2 275.63 ± 394.76）ng/L and the decrease in level of IL-10, respectively were（834.02 ± 271.97）ng/L, （636.83±270.56）ng/L, （682.08±147.85）ng/L, with significant difference. Conclusion Yapa Tang can inhibit lewis lung cancer grow and reduce the number of lung metastases, regulate immune cytokines IL-2 and IL-10, and promote the immunity of tumor-bearing mice.