Gui Zhi Tang, a well-known Chinese herbal formula recorded in the Eastern Han Dynasty, has been widely used to treat exogenous cold for thousands of years. Recent studies showed that Gui Zhi Tang has the effect of regulating the body temperature. In order to better exert the effect of heat production, and protect the vital organs of the body to avoid damage to the cold environment, Jiang Gui Fang (JG) was obtained from clinical utilization in the Department of Traditional Chinese Medicine at General Hospital of Northern Theater Command for many years and has exhibited favorable efficacy. This formula was an empirical formula that was developed based on the research of Gui Zhi Tang, the principles of traditional Chinese medicine and survey a large number of literature. The basis for compatibility of JG was that we selected dried ginger, cassia twig, and licorice in Gui Zhi Tang, which play a major role in the treatment of exogenous cold, and combined with other Chinese medicines, such as pueraria, spatholobus, acanthopanacis cortex, euodiae fructus, codonopsis pilosula.To promote the increase of core temperature of the body and prevent invasion of the major organs from cold environment. We studied the effect of JG on the core body temperature of mice, then explored its regulation of interscapular brown adipose tissue (iBAT) and epididymal white adipose tissue (eWAT) and the possible mechanism. Finally, we determine phytochemical compositions of JG that play a role in heat production.For in vivo studies, we performed a 4-week treatment of JG in acute cold environment at -20 °C and chronic cold exposure at 4 °C. The core temperature, adipose tissue weight, serum parameters, and morphological observation of adipocytes, liver and kidney were measured. Then we investigated the expression levels of adipogenic factors, thermogenic factors and lipoprotein. In vitro, we determined the lipid droplet content, ATP content, and the maximum oxygen consumption of mitochondria.JG treatment promoted core temperature, inhibited eWAT weight, protected kidney, consumed glucose and lipids in Kunming (KM) mice. It also enhanced BAT-associated thermogenic factor uncoupling protein 1 (UCP1) and peroxisome proliferator-actived receptorγ coactivator-1α (PGC1α). The expressions of the lipogenic factor peroxisome proliferate-activator receptor gamma (PPARγ) and lipolytic protein hormone-sensitive triglyceride lipase (HSL) in eWAT were elevated. H&E and immunohistochemistry showed that JG significantly reduced the size of iBAT and eWAT, elevated UCP1 expression levels. In vitro, JG lowered the content of lipid droplets and ATP in brown fat cells. Maximum oxygen consumption capacity of mitochondria and the expression levels of UCP1, PGC1α and silent mating type information regulation 2 homolog 1 (SIRT1) were enhanced after JG treatment.In vivo and in vitro studies showed that JG obviously increased the core temperature of mice by activating iBAT and inducing eWAT browning. The mechanism depended on the PPARγ/SIRT1-PGC1α pathway. Our results will provide a reference for further study of iBAT activation and eWAT browning to heat production.
