不同参数的超声对脂多糖诱导人系膜细胞增殖的影响
Effects of ultrasound with different parameters on lipopolysaccharide-induced human mesangial cell proliferation

摘要：目的 探索不同参数的超声对脂多糖(lipopolysaccharide, LPS)诱导人系膜细胞(human mesangial cell, HMC)增殖的影响，为超声联合微泡和(或)药物治疗肾小球肾炎奠定实验基础。 方法 利用10μg/mL LPS诱导HMC增殖模型，①采用正交设计的方法，将频率为0.8MHz超声的强度(1.48W/cm2，2.56W/cm2，2.79W/cm2)、占空比(1∶4，1∶1，4∶1)和辐照时间(5min，10min，15min)按3因素3水平分成9个实验组，分别作用于LPS诱导的HMC，利用MTT法测定细胞增殖情况，并对这3种超声参数进行极差分析和方差分析；②选定超声占空比为1∶1，对10μg/mL LPS诱导的HMC给予不同强度和辐照时间的超声辐射，观察细胞生长情况。结果 极差分析和方差分析结果显示，这3种超声参数对LPS诱导HMC的增殖生长均有抑制作用，其中占空比是最主要的影响因素，其次为超声强度和辐照时间。当占空比为1∶4时，细胞增殖生长状态均未见明显抑制；当占空比为4∶1时，细胞增殖生长明显受到抑制，其抑制率均达到60.00%以上；当占空比为1∶1时，不同辐照时间与强度组合的超声对LPS诱导HMC增殖的抑制作用不同。结论 不同参数的超声对LPS诱导HMC增殖生长的抑制作用不同，而占空比是体外超声抑制细胞生长最主要的影响因素。
ABSTRACT: Objective To investigate the effects of ultrasound with different parameters on lipopolysaccharide (LPS)-induced human mesangial cell (HMC) proliferation so as to provide the experimental data for ultrasound combined with microbubbles or drug in glomerulonephritis therapy. Methods HMC proliferation model induced by 10μg/mL of LPS was used to determine the effects of ultrasound with different parameters on cellular proliferation. ① Based on three ultrasonic factors including the intensity (1.48W/cm2, 2.56W/cm2 and 2.79W/cm2), duty cycle (1∶4, 1∶1 and 4∶1) and exposure time (5min, 10min and 15min), nine experimental groups were respectively designed by orthogonal methods in LPS-induced HMC proliferation model with different ultrasonic treatments as well as the control group (without any treatment) and the model group (treated by LPS alone) were designed. ② We also examined the effects of ultrasound with different intensity (2.17W/cm2, 2.56W/cm2 and 2.79W/cm2) and exposure time (5min and 10min) at a 1∶1 duty cycle on LPS-induced HMC proliferation. Besides cellular morphology, their growth inhibition rates were evaluated by MTT assay and these three parameters were in turn analyzed in both experiments. Results Range and variance analysis showed that three ultrasonic parameters had inhibitory effects on cellular viability in LPS-induced HMCs, but the duty cycle of the ultrasound was the most important factor. Inhibition on cell growth was not significantly observed in the experimental groups at the duty cycle of 1∶4. However, the cell growth inhibition rates were more than 60.00% in the three experimental groups at the duty cycle of 4∶1. Moreover, the inhibitory effects of ultrasound with different intensity and exposure time at a 1∶1 duty cycle were different on LPS-induced HMC proliferation. Conclusion Ultrasound with different parameters has different effects on LPS-induced HMC growth inhibition, but the duty cycle of the ultrasound is the most