黄芪扶正汤抗肿瘤与免疫调节作用的初步研究
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摘要
目的：
建立Lewis肺癌荷瘤小鼠动物模型,观察黄芪扶正汤及其与顺铂联合使用后的治疗作用和对小鼠免疫功能的影响,初步探讨其可能的作用机制,为黄芪扶正汤在临床上的进一步应用提供实验依据。
方法：
建立肺癌荷瘤小鼠动物模型,随机分为5组：正常小鼠空白对照组、模型对照组、黄芪扶正汤中药组、顺铂化疗组、化疗+中药组,每组7只。皮下接种后第7天,同时开始给药：顺铂组按2mg/kg给药,每3日腹腔注射,共5次。中药组每日10ml/kg.d灌胃一次,共14天。化疗+中药组两药用法同前。对照组采用等量生理盐水灌胃或／和腹腔注射。观察小鼠生活状态,体重变化等,做详细记录。每3天测量肿瘤的最长径(a)、最短径(b),计算肿瘤体积,绘制生长曲线。第15天取血后用颈椎脱臼法处死小鼠,取实体瘤、脾脏、右侧股骨等组织,称重并计算抑瘤率、肺转移抑制率、脾脏指数,计数骨髓有核细胞及外周血白细胞,以MTT法测脾脏T淋巴细胞增殖反应,ELISA法测血清IL-2和IFN-γ含量,评价黄芪扶正汤的抗肿瘤和免疫调节的作用,并初步分析其作用机制。
结果：
1.各组小鼠的一般情况：模型对照组、化疗组小鼠出现进食减少,被毛光泽减弱,精神萎靡,喜欢聚集、行动迟缓现象。中药组、化疗+中药组精神状态一般,觅食积极,无明显消瘦、聚集等现象。各组小鼠体重均有不同程度的增加,化疗组体重增加量最少,有统计学差异(P<0.05)
2.对肿瘤重量和抑瘤率的影响：中药组瘤重较模型对照组减轻(P<0.05)；化疗+中药组的瘤重低于化疗组、中药组(P<0.05)；化疗+中药组的抑瘤率高于化疗组、中药组(P<0.05)；中药与化疗联用表现为疗效相加作用。
3.对肿瘤肺转移的影响：中药组可减少肺转移数,与模型对照组比较有统计学差异(P<0.05),与化疗组无明显差异(P>0.05)；中药加化疗组肺转移数最少(P<0.05),化疗+中药组肺转移抑制率高于化疗组、中药组(P<0.05)。
4.对小鼠外周血白细胞和骨髓有核细胞的影响：与模型对照组相比,化疗组白细胞和骨髓有核细胞数下降(P<0.05),化疗+中药组无明显差异(P>0.05),中药组、空白对照组白细胞数和有核细胞数均升高(P<0.05)。
5.对小鼠脾脏指数的影响：模型对照组与空白对照组相比,脾脏指数降低(P<0.05)；与模型对照组比较,化疗组脾脏指数降低(P<0.05),化疗+中药组、中药组脾脏指数增加(P<0.05)。
6.对脾脏T淋巴细胞增殖反应的影响：与空白对照组相比,模型对照组、化疗组的T淋巴细胞增殖能力下降(P<0.05),而中药组T淋巴细胞增殖能力基本正常(P>0.05)；与化疗组相比,化疗+中药组T淋巴细增殖能力增强(P<0.05)。
7.对小鼠血清IL-2和IFN-y的影响：模型对照组与空白对照组相比,IL-2、IFN-γ下降(P<0.05)；与模型对照组比较,化疗组IL-2、IFN-γ低(P<0.05),化疗+中药组、中药组IL-2、IFN-γ上升(P<0.05)；中药组较化疗+中药组上升(P<0.05)。
结论：
1.黄芪扶正汤可抑制Lewis肺癌小鼠肿瘤生长和转移。
2.黄芪扶正汤对化疗有增效减毒的作用。
3.黄芪扶正汤可提高Lewis肺癌小鼠的脾脏T淋巴细胞增殖,上调血清IL-2、IFN-γ的含量,提高其免疫功能,从而发挥可能的抗肿瘤作用。
Objective:
To study Huangqi Fuzheng Decoction(HFD) and HFD combined with Cisplatin(DDP) on the anti-tumor and immunomodulatory effects by a model of Lewis lung cancer tumor-bearing mice, and then to explore the possible mechanism and provide the experimental basis to tumor therapy with HFD.
Methods:
Lewis lung cancer bearing mice were established and then randomly assigned to one of the following five groups (n=7,each group):normal control group, model group, Huangqi Fuzheng Decoction(HFD) treatment group,Cisplatin(DDP) group, HFD combined with DDP group. Different treatment were respectively given at the 7th day after injection subcutaneously:DDP group was treated with DDP 2mg/kg intraperitoneally every three day for 5 times; HFD group was treated with HFD 10ml/kg orally once daily for 14 days; HFD plus DDP group was adopted the same administration of HFD and DDP; Model control group was treated with the same amount of normal sodium orally and/or intraperitoneally. The healthy condition and body weight of mice were observed and recorded. The long diameter and short diameter of the tumor were measured every three days, then the volume was calculated and the growth curve was drawn. The blood samples were taken and the mice were sacrificed on the 15th day. The neoplasms and spleens were cut down and weighed, and then the parameters of tumor inhibition rate, the inhibitory rate of metastasis on lung surface, the spleen index, the counting of peripheral white blood cells and bone marrow karyocytes were calculated. T lymphocyte proliferation in spleen was detected by MTT assay. The levels of IL-2 and IFN-γin serum were measured by enzyme-linked immunosorbent assay (ELISA). The effects of Huangqi Fuzheng Decoction on the Immunomodulatory and tumor therapy basing on the tumor bearing mice were evaluated.In addition, the probable mechanisms were analyzed.
Results:
1. The effect on the general state of the mice:The mice in model group and DDP group ate small amount of food and had tarnished fur, and they were in low spirit and sluggish in action. The general state of mice in HFD group and DDP plus HFD group was better than that of mice in DDP group. The weight of all the groups was increased more or less, and the increased weight of mice in DDP group was significantly less than that of other groups (P<0.05).
2. The effect on inhibiting tumor:the weight of the tumors in the model group was heavier than that of the HFD group (P<0.05); the weight of the tumors in DDP plus HFD group was lighter than that of other groups (P<0.05), at the same time, the tumor inhibition rate was the highest in all groups(P<0.05).The inhibition of tumor growth presented additive effect that was induced by HFD combined with DDP.
3. The effect on lung metastatic foci:The metastatic foci on lung surface in HFD group were less than those of the model group (P<0.05), and no statistical difference was seen between the HFD group and the DDP group (P>0.05); The metastatic foci on lung surface in the DDP plus HFD group were less than those of other groups (P<0.05). The inhibitory rate of metastasis on lung surface was increased in combined group than that of DDP group and HFD group (P<0.05).
4. The effects on the counting of peripheral white blood cells and bone marrow karyocytes in the mice:Compared to those of the model group, the counting of peripheral white blood cells and bone marrow karyocytes in DDP group were decreased (P<0.05), the HFD group and the normal group were increased (P<0.05), and statistical analysis showed no difference between the model group and the DDP plus HFD group (P>0.05).
5. The effect on spleen index:The spleen index in model group was less than that of the normal control group (P<0.05); Compared to that of the model group, the DDP group had a lower spleen index (P<0.05), while higher spleen index was seen in the DDP plus HFD group and the HFD group.
6. The effect on the proliferation of T-lymphocytes:Compare to that of the normal control group, the model group and the DDP group showed much lower activation effects on the T-lymphocye proliferation (P<0.05), the HFD group did not show significant decrease in the cell proliferation (P>0.05); Compare to that of the DDP group, the DDP plus HFD group had a higher proliferation (P<0.05).
7. The effects on serum IL-2 and IFN-γ:Compared to those of the normal control group, the model group had lower levels of serum IL-2 and IFN-γ(P<0.05).Compared to those of the model group, the DDP group had lower levels of serum IL-2 and IFN-γ(P<0.05),the DDP plus HFD group and HFD group had higher levels of serum IL-2 and IFN-γ(P<0.05).
Conclusions:
1. Huangqi Fuzheng Decoction could inhibit the growth and metastasis of the tumor on on Lewis lung cancer bearing mice.
2. Huangqi Fuzheng Decoction could reduce the side effect of chemotherapy in the mice and increase its efficacy.
3. Huangqi Fuzheng Decoction could increase the proliferation of T-lymphocytes in spleen of mice and up-regulated the levels of serum IL-2 and IFN-γ, thus improves the immunity.This may play a role in anti-tumor effect.
建立Lewis肺癌荷瘤小鼠动物模型,观察黄芪扶正汤及其与顺铂联合使用后的治疗作用和对小鼠免疫功能的影响,初步探讨其可能的作用机制,为黄芪扶正汤在临床上的进一步应用提供实验依据。
方法：
建立肺癌荷瘤小鼠动物模型,随机分为5组：正常小鼠空白对照组、模型对照组、黄芪扶正汤中药组、顺铂化疗组、化疗+中药组,每组7只。皮下接种后第7天,同时开始给药：顺铂组按2mg/kg给药,每3日腹腔注射,共5次。中药组每日10ml/kg.d灌胃一次,共14天。化疗+中药组两药用法同前。对照组采用等量生理盐水灌胃或／和腹腔注射。观察小鼠生活状态,体重变化等,做详细记录。每3天测量肿瘤的最长径(a)、最短径(b),计算肿瘤体积,绘制生长曲线。第15天取血后用颈椎脱臼法处死小鼠,取实体瘤、脾脏、右侧股骨等组织,称重并计算抑瘤率、肺转移抑制率、脾脏指数,计数骨髓有核细胞及外周血白细胞,以MTT法测脾脏T淋巴细胞增殖反应,ELISA法测血清IL-2和IFN-γ含量,评价黄芪扶正汤的抗肿瘤和免疫调节的作用,并初步分析其作用机制。
结果：
1.各组小鼠的一般情况：模型对照组、化疗组小鼠出现进食减少,被毛光泽减弱,精神萎靡,喜欢聚集、行动迟缓现象。中药组、化疗+中药组精神状态一般,觅食积极,无明显消瘦、聚集等现象。各组小鼠体重均有不同程度的增加,化疗组体重增加量最少,有统计学差异(P<0.05)
2.对肿瘤重量和抑瘤率的影响：中药组瘤重较模型对照组减轻(P<0.05)；化疗+中药组的瘤重低于化疗组、中药组(P<0.05)；化疗+中药组的抑瘤率高于化疗组、中药组(P<0.05)；中药与化疗联用表现为疗效相加作用。
3.对肿瘤肺转移的影响：中药组可减少肺转移数,与模型对照组比较有统计学差异(P<0.05),与化疗组无明显差异(P>0.05)；中药加化疗组肺转移数最少(P<0.05),化疗+中药组肺转移抑制率高于化疗组、中药组(P<0.05)。
4.对小鼠外周血白细胞和骨髓有核细胞的影响：与模型对照组相比,化疗组白细胞和骨髓有核细胞数下降(P<0.05),化疗+中药组无明显差异(P>0.05),中药组、空白对照组白细胞数和有核细胞数均升高(P<0.05)。
5.对小鼠脾脏指数的影响：模型对照组与空白对照组相比,脾脏指数降低(P<0.05)；与模型对照组比较,化疗组脾脏指数降低(P<0.05),化疗+中药组、中药组脾脏指数增加(P<0.05)。
6.对脾脏T淋巴细胞增殖反应的影响：与空白对照组相比,模型对照组、化疗组的T淋巴细胞增殖能力下降(P<0.05),而中药组T淋巴细胞增殖能力基本正常(P>0.05)；与化疗组相比,化疗+中药组T淋巴细增殖能力增强(P<0.05)。
7.对小鼠血清IL-2和IFN-y的影响：模型对照组与空白对照组相比,IL-2、IFN-γ下降(P<0.05)；与模型对照组比较,化疗组IL-2、IFN-γ低(P<0.05),化疗+中药组、中药组IL-2、IFN-γ上升(P<0.05)；中药组较化疗+中药组上升(P<0.05)。
结论：
1.黄芪扶正汤可抑制Lewis肺癌小鼠肿瘤生长和转移。
2.黄芪扶正汤对化疗有增效减毒的作用。
3.黄芪扶正汤可提高Lewis肺癌小鼠的脾脏T淋巴细胞增殖,上调血清IL-2、IFN-γ的含量,提高其免疫功能,从而发挥可能的抗肿瘤作用。
Objective:
To study Huangqi Fuzheng Decoction(HFD) and HFD combined with Cisplatin(DDP) on the anti-tumor and immunomodulatory effects by a model of Lewis lung cancer tumor-bearing mice, and then to explore the possible mechanism and provide the experimental basis to tumor therapy with HFD.
Methods:
Lewis lung cancer bearing mice were established and then randomly assigned to one of the following five groups (n=7,each group):normal control group, model group, Huangqi Fuzheng Decoction(HFD) treatment group,Cisplatin(DDP) group, HFD combined with DDP group. Different treatment were respectively given at the 7th day after injection subcutaneously:DDP group was treated with DDP 2mg/kg intraperitoneally every three day for 5 times; HFD group was treated with HFD 10ml/kg orally once daily for 14 days; HFD plus DDP group was adopted the same administration of HFD and DDP; Model control group was treated with the same amount of normal sodium orally and/or intraperitoneally. The healthy condition and body weight of mice were observed and recorded. The long diameter and short diameter of the tumor were measured every three days, then the volume was calculated and the growth curve was drawn. The blood samples were taken and the mice were sacrificed on the 15th day. The neoplasms and spleens were cut down and weighed, and then the parameters of tumor inhibition rate, the inhibitory rate of metastasis on lung surface, the spleen index, the counting of peripheral white blood cells and bone marrow karyocytes were calculated. T lymphocyte proliferation in spleen was detected by MTT assay. The levels of IL-2 and IFN-γin serum were measured by enzyme-linked immunosorbent assay (ELISA). The effects of Huangqi Fuzheng Decoction on the Immunomodulatory and tumor therapy basing on the tumor bearing mice were evaluated.In addition, the probable mechanisms were analyzed.
Results:
1. The effect on the general state of the mice:The mice in model group and DDP group ate small amount of food and had tarnished fur, and they were in low spirit and sluggish in action. The general state of mice in HFD group and DDP plus HFD group was better than that of mice in DDP group. The weight of all the groups was increased more or less, and the increased weight of mice in DDP group was significantly less than that of other groups (P<0.05).
2. The effect on inhibiting tumor:the weight of the tumors in the model group was heavier than that of the HFD group (P<0.05); the weight of the tumors in DDP plus HFD group was lighter than that of other groups (P<0.05), at the same time, the tumor inhibition rate was the highest in all groups(P<0.05).The inhibition of tumor growth presented additive effect that was induced by HFD combined with DDP.
3. The effect on lung metastatic foci:The metastatic foci on lung surface in HFD group were less than those of the model group (P<0.05), and no statistical difference was seen between the HFD group and the DDP group (P>0.05); The metastatic foci on lung surface in the DDP plus HFD group were less than those of other groups (P<0.05). The inhibitory rate of metastasis on lung surface was increased in combined group than that of DDP group and HFD group (P<0.05).
4. The effects on the counting of peripheral white blood cells and bone marrow karyocytes in the mice:Compared to those of the model group, the counting of peripheral white blood cells and bone marrow karyocytes in DDP group were decreased (P<0.05), the HFD group and the normal group were increased (P<0.05), and statistical analysis showed no difference between the model group and the DDP plus HFD group (P>0.05).
5. The effect on spleen index:The spleen index in model group was less than that of the normal control group (P<0.05); Compared to that of the model group, the DDP group had a lower spleen index (P<0.05), while higher spleen index was seen in the DDP plus HFD group and the HFD group.
6. The effect on the proliferation of T-lymphocytes:Compare to that of the normal control group, the model group and the DDP group showed much lower activation effects on the T-lymphocye proliferation (P<0.05), the HFD group did not show significant decrease in the cell proliferation (P>0.05); Compare to that of the DDP group, the DDP plus HFD group had a higher proliferation (P<0.05).
7. The effects on serum IL-2 and IFN-γ:Compared to those of the normal control group, the model group had lower levels of serum IL-2 and IFN-γ(P<0.05).Compared to those of the model group, the DDP group had lower levels of serum IL-2 and IFN-γ(P<0.05),the DDP plus HFD group and HFD group had higher levels of serum IL-2 and IFN-γ(P<0.05).
Conclusions:
1. Huangqi Fuzheng Decoction could inhibit the growth and metastasis of the tumor on on Lewis lung cancer bearing mice.
2. Huangqi Fuzheng Decoction could reduce the side effect of chemotherapy in the mice and increase its efficacy.
3. Huangqi Fuzheng Decoction could increase the proliferation of T-lymphocytes in spleen of mice and up-regulated the levels of serum IL-2 and IFN-γ, thus improves the immunity.This may play a role in anti-tumor effect.
引文
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