## 3 Results
### Effects of GBT and Its Components, ALRP and ZOR, on C-Fos Expression in the Mouse Hypothalamus  
To determine the active component responsible for the thermoregulatory effects of GBT, we investigated the expression of c-fos in the mouse hypothalamus using additional comparative experimental set (Figure 3A). C-fos expression elevated under the heat stress treatment (198 \(\pm\) 15% of control) were significantly decreased under GBT, ZOR, and ALRP treatments (120 \(\pm\) 23%, 84.1 \(\pm\) 9.2%, and 75.9 \(\pm\) 4.6%, respectively). Among them, ALRP treatment showed the most suppressive effects against heat stress-induced c-fos levels in the mouse hypothalamus. Based on these results, we identified ALRP as the active ingredient of GBT.  
To evaluate the effects of ALRP on heat stress-induced c-fos levels, we examined c-fos expression in the mouse hypothalamus (Figure 3B). The protein levels of c-fos in the mouse hypothalamus significantly increased in the heat-stressed vehicle-treated group (155 \(\pm\) 16% of control) than in the control group. However, ALRP treatment decreased c-fos levels, starting at 30 mg/kg/day or more. As below, the protein levels of c-fos in ALRP treated group (30, 100, 300 and 1000) were significantly decreased, compared to the corresponding protein level of the heat stress group (91.8 \(\pm\) 8.3%, 78.5 \(\pm\) 3.2%, 88.4 \(\pm\) 12.3% and 76.4 \(\pm\) 6.9%, respectively).  
Effects of ALRP on Heat Stress-Induced Production of Inflammatory Mediators in the Mouse Hypothalamus  
To determine the effects of ALRP on heat stress-induced inflammatory cytokines in the hypothalamus, we examined the expression levels of IL-9, IL-13, PGE1, and PEG2. Levels of IL-9 significantly increased in the hypothalamic homogenates of mice subjected to heat stress (168 \(\pm\) 3% of control). However, IL-9 expression significantly decreased in the ALRP-treated groups at 300 and 1000 mg/kg/day than in the heat stress group (147 \(\pm\) 3% and 142 \(\pm\) 3%, respectively; Figure 4A). Moreover, IL-13 expression significantly increased in the heat stress group than in the control group (164 \(\pm\) 1% of control). The IL-13 levels significantly decreased in all of the ALRP-treated groups than in the heat stress group, showing a dose-dependent tendency (155 \(\pm\) 1%, 156 \(\pm\) 1%, 139 \(\pm\) 1%, and  
Figure 3: Suppressive effect of GBT and its constituents on heat stress-induced c-fos expression. Quantification of densitometric analyses was performed by measuring the expression ratios of c-fos/\(\beta\)-actin (n = 6 per group) in the lysates of hypothalamus for investigating the effects of GBT and its ingredients, ZOR and ALRP (**A**) and ALRP in a dose-dependent manner (**B**). Values are expressed as means \(\pm\) SEM. * \(p\) < 0.05 and *** \(p\) < 0.001 as compared with the control group. # \(p\) < 0.05, *** \(p\) < 0.01 and *** \(p\) < 0.001 as compared with the heat-stressed vehicle-treated group. GBT; Geongangbuja-tang decoction. ZOR; Zingiber officinal Radix. ALRP; Aconiti Lateralis Radix Preparata.  
134 \(\pm\) 1%, respectively; Figure 4B). PGE2 is abundantly produced in the brain in response to inflammation and mimics stress responses, whereas PGE1 plays an antagonistic role as an anti-inflammatory agent. To evaluate the effects of ALRP on PGE levels in the hypothalamus, we examined the ratio of PGE2 to PGE1 using ELISA, and found it to be significantly upregulated after heat exposure (113 \(\pm\) 3% of control); however, this ratio was significantly restored by ALRP at 300 mg/kg/day (100 \(\pm\) 5%, Figure 4C).