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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (5): 584-590.doi: 10.3736/jcim20120515

• Original Experimental Research • Previous Articles     Next Articles

Effects of Chinese herbal medicine Feiyanning Decoction on the ratio of CD4+CD25+ regulatory T cells and expression of transcription factor Foxp3 in mice bearing Lewis lung carcinoma

Jing Guo, Ju-yong Wang, Zhan Zheng, Qing Wang, Chang-sheng Dong#br#   

  1. Institute of Oncology of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Received:2011-09-06 Accepted:2011-11-23 Online:2012-05-20 Published:2018-06-15

Objective: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on the ratio of CD4 +CD25 + regulatory T cells and expression of transcription factor Foxp3 in mice with Lewis lung cancer.

Methods: Thirty-two male wild-type C57BL/6 mice aging from 6 to 8 weeks were inoculated with Lewis lung cancer cells to establish the tumor-bearing model of Lewis lung carcinoma and were randomly divided into model group, Chinese medicine group, chemotherapy group and Chinese medicine combined with chemotherapy group. After intervention for 14 d with corresponding drugs, behaviors, physical signs and changes of feed consumption of the mice were observed. All mice were sacrificed after drug treatment, and tumors and organs were removed to weigh and calculate organ indexes (lung index, spleen index and thymus index). The percentages of CD4 +CD25 + regulatory T cells in the thymus, spleen and tumor were determined by flow cytometry. The expression of Foxp3 mRNA in the thymus, spleen and tumor tissues was detected by quantitative real-time polymerase chain reaction.

Results: Compared with those in the model group, the mice in the Chinese medicine group showed significant reductions in spleen and thymus indexes and tumor weight, and elevation in the body weight without tumor (P<0.05). The numbers of CD4 +CD25 + regulatory T cells in spleen, thymus and tumor were lower in the Chinese medicine group than in the model group (P<0.05). The expression of Foxp3 mRNA in spleen, thymus and tumor was significantly down-regulated in the Chinese medicine group compared with the model group (P<0.05). There were no significant differences in CD4 +CD25 + regulatory T cell ratio and Foxp3 mRNA expression between the Chinese medicine combined with chemotherapy group and the chemotherapy group.

Conclusion: Feiyanning Decoction can enhance the antitumor immune response and thus play a role in antitumor therapy by reducing the ratio of CD4 +CD25 + regulatory T cells and down-regulating the expression of Foxp3 mRNA.

Key words: Lewis lung cancer, regulatory T cells, Foxp3, drugs, Chinese herbal, mice

Table 1

Primer sequences and the product lengths for PCR analysis"

Target gene Forward primer and reverse primer Fragment length (bp) Annealing temperature (℃) Cycle
Foxp3 F: 5′-TAGTTCCTTCCCAGAGTTCTTCC-3′ 382 65.0 40
β-actin F: 5′-CACCCGCGAGTACAACCTTC-3′ 207 60.4 40

Table 2

Dynamic changes in daily consumption of food of mice"

Group n Consumption of food of each group (g)
Day 0 Day 3 Day 7 Day 11 Day 15
Model 8 18.66 18.40 23.94 21.17 20.37
Chinese medicine 8 18.07 18.45 23.03 22.99 22.94
Chemotherapy 8 18.57 18.19 13.50 11.15 8.22
Combined treatment 8 18.55 18.35 15.82 13.18 13.13

Table 3

Dynamic changes in body weight of mice (x±s)"

Group n Body weight (g)
Day 0 Day 3 Day 7 Day 11 Day 15
Model 8 18.62±0.65 19.09±0.66 19.99±0.71 20.42±0.77 20.45±0.70
Chinese medicine 8 18.55±0.80 19.12±0.90 19.74±0.83 20.23±0.82 20.71±0.80
Chemotherapy 8 18.42±0.54 18.01±0.49 16.81±0.38 15.99±0.33 15.39±0.50
Combined treatment 8 18.47±0.56 18.15±0.62 17.55±0.55 16.41±0.86 15.55±0.58

Table 4

Changes in body weight, body weight without tumor, tumor weight and inhibition rate of tumor of mice"

Group n Body weight
(x±s, g)
Body weight without tumor
(x±s, g)
Tumor weight
(x±s, g)
Inhibition rate of tumor
Model 8 20.45±0.70 16.29±0.79 4.16±0.33
Chinese medicine 8 20.71±0.80 17.42±0.82** 3.29±0.19** 20.91
Chemotherapy 8 15.39±0.50 13.43±0.58 1.95±0.34 53.13
Combined treatment 8 15.55±0.58 14.05±0.64△△ 1.51±0.20 63.70

Table 5

Lung, spleen and thymus indexes of mice (x±s)"

Group n Index of organs (mg/g)
Lung Spleen Thymus
Model 8 11.01±1.26 9.54±0.75 4.76±0.4
Chinese medicine 8 10.59±1.31 8.22±0.64** 4.12±0.29*
Chemotherapy 8 12.40±1.13 5.63±0.78 2.95±0.32
Combined treatment 8 11.54±1.32 6.31±0.34 3.36±0.26

Table 6

Ratio of CD4+CD25+ Treg cells in spleen, thymus and tumor tissue of mice (x±s)"

Group n Ratio of CD4+CD25+ Treg cells (%)
Spleen Thymus Tumor
Model 3 25.23±0.96 23.23±0.61 27.28±0.94
Chinese medicine 3 22.12±1.12* 20.46±1.29* 21.85±0.92**
Chemotherapy 3 20.45±1.05 20.35±1.05 20.88±0.90
Combined treatment 3 18.36±0.84 17.26±0.79 18.43±0.79

Table 7

Expression of Foxp3 mRNA in spleen, thymus and tumor tissue of mice (x±s)"

Group n Foxp3+ mRNA
Spleen Thymus Tumor
Model 3 1.40±0.09 1.37±0.05 1.43±0.04
Chinese medicine 3 1.13±0.05* 1.19±0.04* 1.25±0.08*
Chemotherapy 3 1.06±0.08 1.07±0.07 1.05±0.08
Combined treatment 3 1.03±0.08 1.02±0.09 1.02±0.08
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