Effect Of Berberine On Inflammatory Cytokines Induced By Lipopolysaccharide In THP-1Cells

[Background]Diabetes in our country has become a serious public health problem, the incidence of diabetes has reached to9.7%by2007. Diabetes is the ministry of health "health China2020strategic planning" is listed as one of three big one of the key of the prevention and control of chronic non-communicable diseases. According to a recent study type2diabetes mellitus is not only a kind of metabolic disease. Subclinical chronic inflammation is also a common feature in the natural course of diabetes. And inflammatory factors such as IL-1beta, IL-6, IL-8and TNF alpha, and popular and the onset of diabetes and diabetic complications, especially closely related to cardiovascular events.[Objective]To observing the dose dependent effect of Berberine (Berberine, BBR) on Lipopolysaccharide (Lipopolysaccharide, LPS) induced inflammatory cytokines express in THP-1cells, and to explore the anti-inflammatory effect of BBR.[Methods]1.Cell culture:The THP-1cells were cultured in5%CO2-95%air at37℃in RPMI-1640complete medium containing10%fetal bovine serum(FBS).2. Experimental group:A:Effect of different BBR concentration on the cell viability in THP-1cells.(1) The control group:RPMI-1640complete medium group;(2) Met group:different concentration (BBR5、10、20、50μmol/L);B:Effect of different BBR concentration on the expression of inflammatory factor by LPS in THP-1cells.(1) The control group:RPMI-1640complete medium group;(2) LPS group:cells were cultured in RPMI-1640complete medium containing1μg/mL LPS;(3) BBR group:cells were cultured in RPMI-1640complete medium containing1μg/mL LPS+different BBR concentration (BBR5、10、20μmol/L) collecting cells and culture supernatant after cultured for6h、24h and48h.3. Experimental methods:(1) The survival rate of THP-1cells was measured by Trypan Blue;(2) THP-1cell viability was detected by LDH release assay;(3) The contents of inflammatory factors (TL-1β, IL-6, IL-8and TNF-a) in the culture were measured by ELISA;[Results]1. The survival rate of THP-1cells:the proportion in living cells are more than95%.2. THP-1cells viability:The effects of BBR on the THP-1cell survival rate:When the concentration of BBR was less than10μmol/L, the viability of THP-1cell were more than90%all along and no difference was found compared with the control group (P>0.05). The viability of THP-1cell were less than90%when20μmol/L BBR for48h. When the50μmol/L BBR concentration action for6h、24h or48h, the viability of THP-1cell was less than90%, and the cell number was decreased statistically compared with the control group (P<0.01).3. The levels of inflammatory factor:THP-lcells can only secrete a lower level of IL-1β, IL-6, IL-8and TNF-α without LPS. But under the effect of LPS stimulation, the inflammatory factor levels increased significantly.BBR can dose-dependently decrease the release of IL-1β, IL-6, IL-8and TNF-α from THP-1cells.20μmol/L of BBRcould decrease the secretion of IL-1β, IL-8and TNF-α significantly (P<0.05). at6hours,20μmol/L of BBR could decrease the secretion of IL-1β, IL-8and TNF-α significantly compared with LPS group (P<0.05). At24hours, the secretion of IL-8and TNF-α decreased significantly in20μmol/L of BBR+LPS group compared with LPS group (P<0.05). At48hours, the secretion of TNF-a decreased significantly (F=92.625,P<0.05) in5μmol/L BBR+LPS group, the secrection of IL-1β、IL-6and TNF-α decreased respectively(P<0.05) in10μmol/L BBR+LPS group, and the secretion of IL-1β, IL-6and TNF-a decreased in20μmol/L BBR+LPS group compared with which in LPS group (P<0.05).[Conclusions]BBR can decrease the secretion of inflammatory cytokines in THP-1cells induced by LPS in a dose-dependent manner, suggesting that BBR has certain anti-inflammatory effects.