Study Of The Effect Of Gallnut Water Extract In Periodontal Inflammation And Its Molecular Mechanism

Periodontal disease is a very common oral disease and it frequently occurs among people. During the development of inflammatory perioodontal diseases, lipopolysaccharide (LPS) secreted by putative periodontopathic bacteria not only injuries periodontal tissue directly, but also induces the production of mang kinds of cytokines and inflammatory mediators, thus causing the developmentment of periodoontal inflammation and the destruction of periodontal tissue indirectly. Therefore, the role that LPS is palying in this disease has caught more attention. Previous research has proved that gallnut water extract has a protective effect on the destruction of PDLC by LPS, and it also can inhibit the activity of IL-1 from monocyte induced by LPS. However, the concrete mechanism of gallnut's is still unclear. The aim of this study is to observe the inhibitory effect of gallnut water extract on periodontal inflammation and its effect on cellular singal passway so as to explore the concrete mechanism of gallnu's anti-inflammatory effect.The first part: effect of gailnut water extract on periodontal inflammation1. The effect of gallnut water extract on the IL-6 activity induced by LPS RIA (radio immunoassay) was used to assess the effect of gallnut waterextract on the activitiy of IL-6 in the supernatant after monocyte and HGF was stimulated by LPS. The results showed that gallnut water extract of five different concentration significantly inhibited the activity of IL-6 from HGF and monocyte induced by LPS in a dose-dependent manner. The activity of IL-6 was also decreased after the only administration of gallnut, thus suggesting that gallnut destroyed IL-6 itself and its producing process as well as LPS activity. We concluded that gallnut water extract exerted anti-inflammatory effect through inhibition of the production of inflammatory cytokine from prriodontal cells.2. The effect of gallnut water extract on DNA synthesis and cell cycle of HGF with the inhibitory effect of LPSFlow cytometry (FCM) was used to examine the effect of gallnut water extract on the DNA synthesis and cell cycle of HGF sfter stimulation with LPS. The results showed that The percentage of human gingival fibroblast in Gl phase during exponential growth was increased by LPS, while the percentage of human gingival fibroblast in S phase was decreased. Extracts from Chinese Nutgalls ameliorated this phenomenon. We concluded that Extracts from Chinese Nutgalls can significantly ameliorate the inhibitory effect on the proliferation of human gingival fibroblast induced by LPS, thus suggesting the anti-inflammatory and protective effect of Chinese Nutgalls on periodontal tissue.The second part: the molecular mechanism of the anti-inflammatory effect of gallnut water extract in periondontal inflammation.1. The effect of gallnut water extract on CD 14 expression on monocyte induced by LPSFlow cytometry (FCM) was used to examine the effect of gallnut water extract on CD14 expression on monocyte induced by LPS. The results showedthat after administration with extracts from Chinese Nutgalls, expression of Monocyte CD14 significantly decreased and the percentage of cells expressing CD14 decreased too. We concluded that Extracts from Chinese Nutgalls can inhibit the expression of Monocyte CD 14, thus influencing the combining of LPS and CD 14 and blocked the LPS singal passway.2. The effect of gallnut water extract on raise of [Ca2+] in cells induce by LPS Laser scanning confocal microsopy (LSCM) was used to explore the effectof gallnut water extract on the raise of [Ca2+] in cells induce by LPS. The results showed that [Ca2+] in cells raised after the stimulation of LPS, while gallnut inhibited this effect. We concluded that gallnut water extract influenced the signal transduction in cells through inhibition of raise of [Ca2+] in cells, thus exerting anti-inflammatory effect.3. The effect of gallnut water extract on p38 MAPK activition induced by LPS Western-blots was used to explore the effect of gallnut water extract o...