Clinical And Basic Research On Immunomodulation Of BuZheng Yiqi Xiaozheng Decoction On Lung Cancer Based On Th17/Treg Imbalance

Aims:This study evaluated the immunomodulatory function of BuZheng Yiqi XiaoZheng on Lewis tumor-bearing C57BL/6 mice by analyzing the characteristics of Th17/Treg imbalance of the patients with lung cancer, to provide a scientific basis for its clinical trial application.Methods:1. In this part of the study, serum samples of patients with lung adenocarcinoma and lung squamous cell carcinoma were collected and used to determine the cytokines RORyt, Foxp3, TGF-β1, IL-6 and IL-10 which are associated with Th17 and Treg by ELISA.2. On the basis of clinical evaluation on the correlation of Th17/Treg imbalance with tumor stage of lung cancer, Lewis tumor-bearing C57BL/6 mice was used as a in vivio model, combining with high frequency color Doppler ultrasound test, enzyme linked immunosorbent assay (ELISA) and immunohistochemistry, and other methods, to evaluate the tumor inhibition and immunomodulatory of Yiqi Jianpi xiaozheng in Lewis tumor-bearing C57BL/6 mice.3. In this part of the study, the anti-tumor efficacy and immunomodulatory effects of SB were evaluated by Doppler ultrasound test, Western blot, ELISA and immunohistochemistry. The results showed that the alkaloids and flavonoids of SB could reduce the levels of RORyt, IL-17, IL-6, Foxp3, IL-10 and TGF-β1, while increase the levels of IL-2 and IFN-y, in serum and spleen of Lewis tumor-bearing C57BL/6 mice, compared with model mice.Results:1. The correlation between the staging of clinical patients with lung cancer and Th17, Treg.The results showed that the levels of RORyt, IL-6, Foxp3, TGF-β1 and IL-10 of lung adenocarcinoma and lung squamous carcinoma group were significantly increased compared with the healthy group. There is a positive correlation between RORyt/Foxp3 and tumor stage because of higher tumor stage with higher RORyt/Foxp3 ratio. There is a positive correlation between IL-10 and RORyt/Foxp3 because of higher RORyt/Foxp3 ratio whereas higher IL-10 in higher tumor stage. The level of IL-6 was higher in higher tumor stage with a positive correlation between IL-6 and RORyt/Foxp3. In adenocarcinoma and squamous carcinoma groups, TGF-β1 and RORyt/Foxp3 ratio was positively correlated with higher TGF-β1.level in higher tumor stage. The results showed that the stages of lung cancer were associated with the imbalance of RORyt/Foxp3. We can improve immune function of patients with lung cancer by regulating and controling imbalance of immune system Thl7/Treg. Both Thl7cells and Treg cells in peripheral blood of the patients with lung cancer increased with a positive linear correlation between them, however, the ratio of Th17/Treg cells were increased significantly, compared with healthy people. This indicated that there was an imbalance of Treg/Thl7 ratio in patients with lung cancer, especially, with the relatively significant increase of Th17 cells. Improving the immune function of patients with lung cancer can be archived by regulating the imbalance of Th17/Treg.2. Immunomodulatory of BuZheng Yiqi XiaoZheng on Lewis tumor-bearing C57BL/6 mice.The results showed that Yiqi Jianpi xiaozheng could significantly inhibit the tumor volume and tumor weight of Lewistumor-bearing C57BL/6 mice, enhance thymus index and spleen index, reduce the levels of RORyt, IL-17, IL-6, Foxp3, IL-10 and TGF-β1 in serum of tumor-bearing mice, and increase the levels of IL-2 and IFN-γ (p< 0.05). These results indicated that BuZheng Yiqi XiaoZheng could significantly inhibit tumor growth of lung cancer by regulating the imbalance of Th17/Treg.3. Immunomodulatory activity of Scutellaria barbata D. Don (SB) on Lewis tumor-bearing C57BL/6 mice.The results showed that the alkaloids and flavonoids of SB could reduce the levels of RORyt, IL-17, IL-6, Foxp3, IL-10 and TGF-β1, while increase the levels of IL-2 and IFN-y, in serum and spleen of Lewis tumor-bearing C57BL/6 mice, compared with model mice. The results suggested that the anti-tumor efficacy of SB was associated with the regulation of SB on the imbalance of Th17/Treg.Conclusions:1. With the progress of lung cancer stages, the imbalance of Th17/Treg appear more obvious. The underlying mechanism may be associated with cytokines produced by the tumor itself. The ratio of RORyt/Foxp3 can be used as early biomarkers and the reference for prognosis of treatment.2. BuZheng Yiqi XiaoZheng could significantly inhibit the growth of lung cancer and regulate the imbalance of Th17/Treg. Regulation of immune function has become an important measure for lung cancer treatment.3. SB in BuZheng Yiqi XiaoZheng could significantly inhibit the growth of lung cancer and regulate the imbalance of Th 17/Treg.