臭牡丹提取物对H22荷瘤小鼠体内抑瘤作用及机制研究

Antitumor effect and mechanism of Clerodendron Bungei extract on H22 tumor-bearing mice

目的研究臭牡丹提取物对H22实体移植瘤的抑制作用及其机制。方法皮下接种H22瘤株建立小鼠H22实体移植瘤模型。按照体重将荷瘤鼠随机分为5组:模型组、对照组和低、中、高3个剂量实验组,每组12只。模型组给予0. 9%NaCl灌胃;对照组给予5-氟尿嘧啶(5-Fu) 25 mg·kg^-1腹腔注射;低、中、高3个剂量实验组分别灌胃臭牡丹提取物0. 55,1. 09,2. 18 g·kg^-1,1次/天,连续14 d。称重法检测小鼠肿瘤重量;以流式细胞术检测外周血CD4+和CD8+T淋巴细胞;以免疫印迹法检测磷脂酰肌醇3-激酶(PI3K)和磷酸蛋白激酶(p-Akt)蛋白的表达水平。结果模型组、对照组和低、中、高3个剂量实验组肿瘤重量分别为(4. 13±0. 51),(1. 16±0. 15),(3. 09±0. 49),(2. 68±0. 43)和(1. 98±0. 16) g,对照组和中、高2个剂量实验组与模型组相比,差异均有统计学意义(均P <0. 01)。上述这5组PI3K蛋白相对表达量(灰度值)分别为0. 51±0. 06,0. 25±0. 04,0. 47±0. 07,0. 32±0. 04和0. 18±0. 03;上述这5组P-Akt蛋白相对表达量(灰度值)分别为0. 38±0. 06,0. 16±0. 02,0. 37±0. 04,0. 33±0. 04和0. 13±0. 01,对照组和高剂量实验组与模型组相比,差异均有统计学意义(均P <0. 01)。上述这5组CD4+/CD8+比值分别为1. 84±0. 12,1. 65±0. 11,3. 48±0. 15,3. 56±0. 23和3. 77±0. 18,3个剂量实验组与模型组相比,差异均有统计学意义(均P <0. 01)。结论臭牡丹提取物能够抑制H22实体移植瘤生长,其机制可能与抑制PI3K/Akt信号通路和调节机体免疫功能有关。

Objective To study the inhibitory effect of Clerodendron Bungei extract on H22 solid xenograft and its mechanism. Methods A mouse H22 solid xenograft model was established by subcutaneous inoculation of H22 tumor strain. The tumor-bearing mice were randomly divided into five groups: model group,control group,and experimental-L,experimental-M and experimental-H groups,with 12 mice in each group. The model group was given normal saline;control group was given 5-fluorouracil( 5-Fu) 25 mg·kg^-1 intraperitoneally;experimental-L,experimental-M and experimental-H groups were respectively administered with Clerodendron Bungei extract 0. 55, 1. 09, 2. 18 g·kg^-1,1 times/day for 14 days. The tumor mass was measured by weighing method. The peripheral blood CD4+and CD8+T lymphocytes were detected by flow cytometry. The expression levels of Phosphatidylinositol 3 kinase( PI3K)/Serine threonine protein kinase( Akt) protein were detected by Western blot. Results The tumor weights of model group,control group and experimental-L,experimental-M and experimental-H groups were( 4. 13 ± 0. 51),( 1. 16 ± 0. 15),( 3. 09 ± 0. 49),( 2. 68 ± 0. 43),( 1. 98 ± 0. 16) g;comparison between control group and experimental-M,experimental-H groups with model group,the difference was significant( P < 0. 01). The relative expression levels of PI3K protein( grey value) in the above 5 groups were 0. 51 ± 0. 06,0. 25 ± 0. 04,0. 47 ± 0. 07,0. 32 ± 0. 04 and 0. 18 ± 0. 03;the relative expression levels of P-Akt protein( grey value) in the above 5 groups were 0. 38 ± 0. 06,0. 16 ± 0. 02,0. 37 ± 0. 04,0. 33 ± 0. 04 and 0. 13 ± 0. 01;comparison between control group and experimental-H group with model group,the difference of the factors were significantly( all P < 0. 01). The CD4+/CD8+ ratios in the above 5 groups were 1. 84 ± 0. 12,1. 65 ± 0. 11,3. 48 ± 0. 15,3. 56 ± 0. 23,3. 77 ± 0. 18;comparison between three dose experimental groups and the model group,the difference was statistically significant( P < 0. 01). Conclusion The Clerodendron Bungei extract can inhibit the growth of H22 solid xenografts,and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway and regulation of immune function.