Study Of The Mechanisms Of DANCE As A Pathological Response Of Cells In Vitro And In Vivo

Objective: Our previous studies found that DANCE(Density Alteration in Non-physiological Cells) was a common cellular response in lifes, including cultured human cell lines, mouse tissue cells, Escherichia coli and candida albicans, etc. This paper was designed to study DANCE as a pathological response of cells in vitro and in vivo by using cultured human cells, E. coli, C. albicans, and adrenaline-induced mouse model of acute pulmonary edema, and thereby to study the biochemical mechanisms of DANCE in vivo.Methods: Determine the DANCE response of cells, E. coli and C. albicans after the culture conditions were changed and after the cells/microorganisms were incubated with drugs; The liver and lung tissues of normal KM mice were separated by DSGC into bands of tissue cells; The DSGC bands were stained with HE; A mouse model of acute pulmonary edema was constructed via intraperitoneal injection of adrenaline hydrochloride, and the DANCE response in the liver and the lung tissue cells was determined by DSGC; The mouse lung coefficients were determined; Slides of mouse lung tissue sections were prepared; Anthrone colorimetry was employed to determine the glycogen contents of the hepatic tissues; Quantitative Real-time PCR was performed to determine the m RNA expression levels of IL-1β and IL-6 in the mouse liver cells after injection of adrenaline at different doses and with different durations. Analysis was carried out for the relationship between the results and the incidence of DANCE in the hepatic and lung tissue cells.Results:After DSGC, K562 cells, E. coli and C. albicans were separated into several bands of cells with different intracellular densities. Change of culture conditions and incubation with drugs significantly caused changes in the number and cell quantity of the DSGC bands; After staining with HE dyes, the DSGC bands from the lung and the liver homogenates were microscopically found to be mainly composed of liberated cells and tissue fragments; The DANCE response in the lungs and the liver showed significant dose- and time- dependence on the drug. DANCE incidence in the lungs showed two peaks at 2 and 6 hrs, while in the liver, two peaks at 1 and 8 hrs, respectively, after injection of adrenaline. Pearson analysis showed that the incidence of DANCE in the lungs and the liver was significantly correlated with the dosage of adrenaline; The incidence of DANCE in the lungs was found to be significantly correlated with the lung coefficients in the experimented mice; Severe accumulation of blood cells was microscopically observed in the slides of the lung tissues from the mice with DANCE in the lungs; An inversed peak of the liver glycogen content in the mice with acute pulmonary edema was observed at 1 hr, while a normal peak was found at 8 hrs after administration of adrenaline; Significant correlation was found between the DANCE incidence and the m RNA levels of IL-1β and IL-6 in the liver cells. The relative expression levels of IL-1β and IL-6 m RNAs in the liver reached a peak at the time point of 1 hr after injection of the drug.Conclution: The in vitro experiments domonstrated that change of culture conditions and incubation with drugs may cause DANCE response in cells and microorganisms, and DANCE occurs as a reflex of pathophysiological changes in the cells and the microorganisms. Acute pulmonary edema induced by over-dosed adrenaline may cause DANCE in the liver and the lungs, and the incidence of DANCE in the two organs is closely related to the dose and action duration of the drug. Since it was accompanied by increase of m RNA levels of IL-1β and IL-6 in the liver cells, DANCE in the liver and the lungs was suggested to be a pathological response to the drug induced acute pulmonary edema. Therefore, the mechanisms for DANCE in the liver and the lungs caused by adrenalin-induced acute pulmonary edema could be the combination of the drug induced and cytokine mediated stress and inflammatory responses in the body, change of the mode of metabolism in the liver cells(such as metabolism of the liver glycogen), squeezing on the lung tissue cells due to the high pressure inside the lungs, and accumulation of blood cells in the lung tissues due to congestion of the pulmonary circulation.