ABSTRACT
Objective To investigate the effects of ultrasound combined with microbubbles on intracellular Ca2+ homeostasis in carboplatin ( CBP )-treated A549 cell and its possible mechanisms of inhibiting A549 cell line activity . Methods According to whether SonoVue was used or not ,and the different dose of CBP ,the groups A-F were arranged as the ultrasound(US) group(group A) ,the ultrasound combined with microbubbles ( USMB) group( group B) ,the low dose CBP ( 100 μg/ml) + US group( group C) ,the low dose CBP+USMB group( group D) ,the high dose CBP ( 200 μg/ml)+ US group ( group E) and the high dose CBP+USMB group( group F) .A549 cells were bathed and washed by a calcium-free buffer , loaded with Ca2+ indicator fluo-4 AM . Real-time images were acquired using laser confocal microscopy .The fluorescence intensity of intracellular calcium ion concentration ([Ca2+ ]i) in individual living cell was observed and the calcium overload was analyzed . Results After ultrasound irradiation ,the normalized fluorescence intensity of [Ca2+ ]i increased rapidly ,then returned to a new homeostasis (selected cells in groups A ,B ,E ,F) or experienced a second calcium oscillation ( some cells in group C and D) . All the selected cells in group B and some cells in group C and D exhibited superimposed oscillations . The calcium overloading time in group D was longer than those of any other groups . Four cells in group A experienced delayed calcium oscillations . Compared with group A ,the selected cells in other groups exhibited a larger amplitude of calcium oscillation( all P < 0 .05) and the selected cells in group B and D exhibited calcium oscillation for a longer period of time( all P <0 .05) . Conclusions In the calcium-free buffer ,US ,USMB , CBP+ US ,CBP + USMB are direct stimuli of calcium overload in A 549 cells . SonoVue ,CBP ,CBP +SonoVue are all synergistic stimuli of calcium overload in A 549 cells irradiated by ultrasound .US ,USMB and CBP may synergistically induce calcium release from intracellular store sites in A 549 cells . Calcium overload is a possible mechanism of ultrasound combined with microbubbles in assisting CBP chemotherapy .
ABSTRACT
To investigate the effects of ultrasound combined with microbubbles on intracellular Ca2+ homeostasis in carboplatin ( CBP )‐treated A549 cell and its possible mechanisms of inhibiting A549 cell line activity . Methods According to whether SonoVue was used or not ,and the different dose of CBP ,the groups A‐F were arranged as the ultrasound( US) group( group A ) ,the ultrasound combined with microbubbles ( USMB) group( group B) ,the low dose CBP ( 100 μg/ml) + US group( group C) ,the low dose CBP+USMB group( group D) ,the high dose CBP ( 200 μg/ml)+ US group ( group E) and the high dose CBP+USMB group( group F) .A549 cells were bathed and washed by a calcium‐free buffer , loaded with Ca2+ indicator fluo‐4 AM . Real‐time images were acquired using laser confocal microscopy . T he fluorescence intensity of intracellular calcium ion concentration ( [ Ca 2+ ] i ) in individual living cell was observed and the calcium overload was analyzed . Results After ultrasound irradiation ,the normalized fluorescence intensity of [ Ca2+ ] i increased rapidly ,then returned to a new homeostasis ( selected cells in groups A ,B ,E ,F ) or experienced a second calcium oscillation ( some cells in group C and D ) . All the selected cells in group B and some cells in group C and D exhibited superimposed oscillations . T he calcium overloading time in group D was longer than those of any other groups . Four cells in group A experienced delayed calcium oscillations . Compared with group A ,the selected cells in other groups exhibited a larger amplitude of calcium oscillation ( all P ＜ 0 .05 ) and the selected cells in group B and D exhibited calcium oscillation for a longer period of time( all P ＜0 .05) . Conclusions In the calcium‐free buffer ,US ,USMB , CBP+ US ,CBP + USMB are direct stimuli of calcium overload in A 549 cells . SonoVue ,CBP ,CBP +SonoVue are all synergistic stimuli of calcium overload in A 549 cells irradiated by ultrasound .US ,USMB and CBP may synergistically induce calcium release from intracellular store sites in A 549 cells . Calcium overload is a possible mechanism of ultrasound combined with microbubbles in assisting CBP chemotherapy .
ABSTRACT
Objective To investigate the effect of Propranolol and Phentolamine on neurogenic pulmonary ede-ma(NPE)in rats. Methods One hundred and twenty male Wistar rats were randomly divided into 4 groups:the con-trol group(group A),the NPE group(group B),the Propranolol treatment group(group C)and the Phentolamine treatment group(group D),30 cases in each group. Diffuse brain injury was induced in the latter 3 groups. The lung wet/ dry ratio was calculated. HE staining was used to measure the histological changes in the lung tissues. The levels of neuropeptide Y(NPY)and substance P(SP)in the serum and the bronchoalveolar lavage fluid(BALF)were detected by enzyme - linked immunosorbent assay(ELISA). The expressions of NPY and SP in the lung tissues were demonstra-ted by immunohistochemical staining,and immunohistochemical scores(IHS)were measured after scarifying the ani-mals at different time points(0. 5,6. 0 and 24. 0 h after injury). Results Compared with group A,water volume in the lungs in group B increased at 24. 0 h(P ﹤ 0. 05);NPY content in the serum was elevated at 6. 0 h,while that in BALF was elevated at 6. 0 h and 24. 0 h(all P ﹤ 0. 05);SP content in the serum was elevated at 0. 5 h and 6. 0 h(all P ﹤0. 05),while that in BALF was elevated at 0. 5 h(P ﹤ 0. 05);The expression of NPY protein in the lung tissue in-creased at 0. 5 h,6. 0 h and 24. 0 h(all P ﹤ 0. 05),while the level of SP protein increased at 0. 5 h(P ﹤ 0. 05). Com-pared with group B,water volume in the lungs in group C was higher at 6. 0 h and 24. 0 h(all P ﹤ 0. 05);NPY concen-trations in the serum were higher at 6. 0 h and 24. 0 h(all P ﹤ 0. 05),while those in BALF were higher at 0. 5 h, 6. 0 h,and 24. 0 h(all P ﹤ 0. 05);SP concentrations in serum and BALF were higher at 0. 5 h(all P ﹤ 0. 05). The ex-pression of NPY protein increased at 6. 0 h(P ﹤ 0. 05),while the levels of SP protein increased at 0. 5 h,6. 0 h and 24. 0 h(all P ﹤ 0. 05). Compared with group B,the level of NPY in serum in group D was lower at 6. 0 h,and that in BALF was lower at 6. 0 h and 24. 0 h(all P ﹤ 0. 05). The level of SP in serum was lower at 0. 5 h(P ﹤ 0. 05). The ex-pression of NPY protein decreased at 6. 0 h and 24. 0 h(all P ﹤ 0. 05),while the levels of SP protein decreased at 0. 5 h(P ﹤ 0. 05). Conclusions Phentolamine is effective in reducing NPE through reduction of NPY and SP,while propranolol can stimulate the release of NPY and SP to aggravate NPE following traumatic brain injury in rats.