| Objective: Long-term overtraining is likely to lead to sports fatigue,accompanied by imbalance of the body’s immune function and reduced resistance.Zuo Gui Wan can improve the immune function of the body,but there are few studies in the field of exercise immunity,and the mechanism of regulating the imbalance of exercise immune function is not clear.In this paper,we investigated the regulatory effects of Zuo Gui Wan on the Th1/Th2 balance and inflammatory factor levels in exercising rats,and then provided some theoretical basis for the application of Zuo Gui Wan to alleviate exercise fatigue and improve the immunity of the body.Methods: Fifty SD rats(healthy,male)were randomly divided into blank control group(C),exercise control group(E),Zuo Gui Wan in three dose groups-high(ZEH),medium(ZEM)and low(ZEL).After three days of adaptive swimming,rats were subjected to exhaustive swimming exercise with the weight(4% of their own body weight)for 5 days/week for four weeks.The rats were given 2 ml of Zuo Gui Wan Solution 2h before exercise,and the three doses were 8 g/(kg·d),4 g/(kg·d)and 2 g/(kg·d).The spleen index was calculated.The microstructure of the spleen was observed by HE staining.The concentrations of serum C,T,IFN-γ,TNF-α,IL-2,IL-4,IL-6,IL-10 and IL-12 were measured by ELISA.The gene and protein levels of IFN-γ,IL-4,T-bet,GATA-3 were detected by RT-q PCR and WB.Results:(1)Results of spleen morphological changes.The spleen morphology and structure of the rats were obviously destroyed.The splenic red and white marrow of group E were blurred,the red marrow was sparse,the lymph sheath around the central splenic artery was thickened,and the germinal centers were increased.The morphological structure of the spleen was improved in the three dose groups,and the most obvious improvement was observed in the ZEH group,where the boundaries between the red and white marrow were clear,the thickness of the lymphatic sheath around the central splenic artery was moderate,and the number of hair-growing centers was reduced.(2)Statistical results of spleen indices.Compared with group C,the spleen index in group E was highly significant lower(P<0.01),and the spleen index in both ZEH and ZEM groups was significantly higher(P<0.05)compared with group E.However,the differences between the three dose groups were not significant.(3)Serum fatigue index test results.Serum T/C ratio was highly significantly decreased in group E compared with group C(P<0.01);serum T/C ratio was highly significantly increased in all three dose groups compared with group E(P<0.01);T/C ratio was highly significantly increased in both ZEH and ZEM groups compared with E and ZEL group(P<0.01).(4)Results of the measurement of serum cytokine levels.Compared with group C,IL-6,IL-12 levels and TNF-α/IL-10 ratio were highly significantly increased in group E(P<0.01);IL-2 levels were significantly decreased(P<0.05)and serum IFN-γ/IL-4 ratio was highly significantly decreased(P<0.01).Compared with group E,IL-2,IL-12 levels and serum IFN-γ/IL-4 ratio were elevated in all three dose groups,with the most significant elevation in IL-2 levels and serum IFN-γ/IL-4 ratio in ZEH and ZEM groups(P<0.01)and a highly significant elevation in IL-12 in ZEH group(P<0.01).Compared with group E,IL-6 levels were highly significantly decreased in both ZEH and ZEM groups(P<0.01);TNF-α/IL-10 ratios were significantly decreased in all three dose groups,with the most significant decrease in the ZEH group(P<0.01).(5)Splenic Th1/Th2 balance-specific cytokine assay results.Compared with group C,splenic IFN-γ m RNA and IFN-γ/IL-4 ratio in group E were extremely significantly decreased(P<0.01);splenic IFN-γ protein expression in group E was significantly decreased and IL-4 protein expression was significantly increased.Compared with group E,IFN-γ m RNA and IFN-γ/IL-4 ratio were highly significantly increased in group ZEH(P<0.01);spleen IFN-γ protein expression was significantly increased and IL-4 protein expression was significantly decreased in groups ZEH and ZEM.(6)Results of splenic Th1/Th2 homeostasis-specific transcription factors.Compared with group C,splenic T-bet m RNA and T-bet/GATA-3 ratio were highly significantly decreased in group E(P<0.01);compared with group E,both T-bet m RNA and T-bet/GATA-3 ratio were significantly increased in the administered group(P<0.01),and splenic T-bet protein expression was significantly increased and GATA-3 protein expression was significantly decreased.Conclusions:(1)Zuo Gui Wan can improve the morphological structure of the spleen of exhausted rats,increase the splenic index and improve the physiological function of the spleen of exhausted rats.(2)Zuo Gui Wan can relieve the excessive fatigue caused by four weeks of exhaustive exercise in rats and promote the recovery of physical functions.(3)Zuo Gui Wan can promote the release of anti-inflammatory factors,inhibit the increase of pro-inflammatory factors,and prevent systemic inflammatory reactions caused by exhaustive exercise.(4)Zuo Gui Wan can break the Th1 cell-mediated immunosuppression,reverse the Th1/Th2 immune imbalance and reshape the dynamic balance between cellular immunity/humoral immunity in exhausted rats.(5)High-dose Zuo Gui Wan was most effective in improving Th1/Th2 immune imbalance and controlling high inflammation levels. |
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