Takeuchi Hiroto, Ohmori Koji, Kondo Isao, Shinomiya Kaori, Oshita Akira, Takagi Yuichiro, Yoshida Junji, Mizushige Katsufumi, Kohno Masakazu
Second Department of Internal Medicine, Kagawa University School of Medicine, 1750-1, Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0793, Japan.
Radiology. 2004 Mar;230(3):735-42. doi: 10.1148/radiol.2303020897. Epub 2004 Jan 22.
To confirm that BR14 microbubbles (MBs) can be phagocytosed by activated leukocytes, to determine their stability after phagocytosis, and to evaluate how such characteristics influence the fate of neutrophils containing MBs after insonation.
BR14 and human albumin MBs (2 x 10(7)/mL) were incubated with activated human neutrophils (2 x 10(6)/mL) to allow phagocytosis. Deflation rate of the phagocytosed MBs after pulsed insonation (one burst per second for 5 seconds) at 1.8 MHz with peak negative pressure of -540 kPa or -1,340 kPa, lactate dehydrogenase (LDH) leakage, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling stain-positive cell count after insonation were compared between the two agents.
At -540 kPa, phagocytosed MBs remained nearly unchanged for both agents after insonation. At -1,340 kPa, although human albumin MBs were disrupted on the first or second burst, BR14 MBs remained undisrupted. After -540-kPa insonation, a similar number of apoptotic cells appeared in neutrophils containing human albumin and BR14 MBs. At -540 kPa, LDH leakage was limited in human albumin MBs and BR14 MBs. At -1,340 kPa, LDH leakage was significantly increased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). Apoptotic cells were significantly decreased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). LDH leakage was lower and apoptotic cell count was greater in BR14 MB-containing neutrophils than in human albumin MB-containing neutrophils (both P <.01).
Compared with human albumin MBs, BR14 MBs were more stable after phagocytosis with insonation. This stability is associated with less disruption and greater induction of apoptosis in leukocytes after relatively high-pressure insonation in the range for diagnostic use.
确认BR14微泡（MBs）可被活化的白细胞吞噬，确定吞噬后的稳定性，并评估这些特性如何影响超声照射后含MBs的中性粒细胞的命运。
将BR14和人白蛋白MBs（2×10⁷/mL）与活化的人中性粒细胞（2×10⁶/mL）孵育以实现吞噬。比较两种制剂在1.8MHz、峰值负压为-540kPa或-1340kPa的脉冲超声照射（每秒1次脉冲，持续5秒）后吞噬的MBs的缩小率、乳酸脱氢酶（LDH）泄漏情况以及超声照射后末端脱氧核苷酸转移酶介导的dUTP缺口末端标记染色阳性细胞计数。
在-540kPa时，两种制剂超声照射后吞噬的MBs几乎保持不变。在-1340kPa时，虽然人白蛋白MBs在第一次或第二次脉冲时就被破坏，但BR14 MBs仍未被破坏。在-540kPa超声照射后，含人白蛋白和BR14 MBs的中性粒细胞中出现的凋亡细胞数量相似。在-540kPa时，人白蛋白MBs和BR14 MBs中的LDH泄漏有限。在-1340kPa时，人白蛋白MBs和BR14 MBs中的LDH泄漏显著增加（两者与-540kPa相比，P<.01）。人白蛋白MBs和BR14 MBs中的凋亡细胞显著减少（两者与-540kPa相比，P<.01）。含BR14 MBs的中性粒细胞中的LDH泄漏低于含人白蛋白MBs的中性粒细胞，且凋亡细胞计数更高（两者P<.01）。
与人白蛋白MBs相比，BR14 MBs在超声照射吞噬后更稳定。这种稳定性与在诊断用相对高压超声照射后白细胞中较少的破坏和更大的凋亡诱导相关。
