Investigation On The Anti-inflammatory Effect Of Total Flavones Of Abutilon Theophrasti Medic.Leaves In Vitro

Inflammation is the first defense reaction of immune system to harmful stimulus includi-ng infection, injury and so on in living tissue. Inflammation response is complicated and has dual nature, excessive inflammation dues to body damage. Inflammation can be induced by lots of factors. Especially the Lipopolysaccharide (LPS), a common component of the outer membrane of bacteria can serve as a mediator of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) by activating innate immune systems. NF-κB and MAPK can regulate these inflammatory factor expression. Besides, macrophages are also known to play a key role in inflammation and activated macrophages can mediate a series of inflammation response by releasing NO、PGE2、TNF-α and IL-1β. Currently, steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs are mainly used to treat inflammation in clinical practice. But they both have some side effects. Therefore, it is important for many scholars to find and develop new anti-inflammatory drugs. Abutilon theophrasti Medic. Leaves is an annual herb from Abutilon Malvaceae. Abutilon theophrasti Medic. Leaves contains flavonoids, coumarins, phenols, steroids and organic acid compounds and flavonoids is abundant. However, flavonoids compounds has been known to exhibit antioxidant, antibacterial, anticancer, anti-inflammatory, antitum and other important therapeutic activities. In the present research, the anti-inflammatory effects of total flavones of Abutilon theophrasti Medic. Leaves were studied in vitro.In LPS stimulated RAW264.7 cells, the effects of total flavones of Abutilon theophrasti Medic. Leaves on inflammatory mediators (NO, COX-2)、cytokines (TNF-α, IL-6, and IL-1β)、NF-κB and MAPK signaling pathways (p65, IκB, p38α, JNK, ERK1/2) were analyzed by Griess, enzyme-linked immunosorbent assay (ELISA), real-time fluorescence quantification PCR and Western blot.The results showed that total flavones of Abutilon theophrasti Medic. Leaves markedly inhibited LPS-induced NO, COX-2, TNF-α, IL-6 and IL-1β content in vitro. And total flavones of Abutilon theophrasti Medic. Leaves markedly inhibited LPS-induced iNOS, COX-2, p65, p38α, ERK and JNK mRNA expression and increased IκB mRNA expression. Furthermore, total flavones of Abutilon theophrasti Medic. Leaves inhibited the phosphorylation of p65, p38α, JNK and ERK in LPS stimulated RAW264.7 cells and IκB degradation, increased p-IκB expression also. We found that total flavones of Abutilon theophrasti Medic. Leaves can directly inhibit the expression of inflammatory factors through testing the influence of on NO, COX-2, TNF-α, IL-6 and IL-1β in LPS stimulated RAW264.7 cells. We prove that total flavones of Abutilon theophrasti Medic. Leaves can inhibit the activation of LPS-induced NF-κB via stabilizing IκB. The expression of many enzymes and cykines involved in the inflammatory response is also regulated by MAPK, so upstream MAPK may be the target of total flavones of Abutilon theophrasti Medic. Leaves LPS can activate ERK1/2、JNK and p38 MAPK by stimulate macrophages, so we study the effection of total flavones of Abutilon theophrasti Medic. Leaves on MAPK. The results indicate total flavones of Abutilon theophrasti Medic. Leaves can inhibit the level of phosphorylation of ERK1/2、JNK and p38. In conclusion, total flavones of Abutilon theophrasti Medic. Leaves can directly inhibit the expression of inflammatory factors but also inhibit NF-κB and MAPKs.Based on the study of total flavones of Abutilon theophrasti Medic. Leaves anti-inflammatory activation in vitro, we preliminary reveals the anti-inflammatory effects and mechanism of total flavones of Abutilon theophrasti Medic. Leaves, and this study provides a theoretical basis for the treatment of inflammation and establishes a foundation for development of anti-inflammatory drugs.