Department of Epidemiology, College of Public Health, University of Nebraska Medical Center, Omaha, NE 68198-5910, USA.
Respir Res. 2012 Jun 19;13(1):49. doi: 10.1186/1465-9921-13-49.
Haemophilus influenzae infection of the nasal epithelium has long been associated with observations of decreased nasal ciliary beat frequency (CBF) and injury to the ciliated epithelium. Previously, we have reported that several agents that slow CBF also have the effect of activating protein kinase C epsilon (PKCε) activity in bronchial epithelial cells. The subsequent auto-downregulation of PKCε or the direct inhibition of PKCε leads to the specific detachment of the ciliated cells.
Primary cultures of ciliated bovine bronchial epithelial cells were exposed to filtered conditioned media supernatants from non-typeable H. influenzae (NTHi) cultures. CBF and motile points were measured and PKCε activity assayed.
NTHi supernatant exposure significantly and rapidly decreased CBF in a dose-dependent manner within 10 minutes of exposure. After 3 hours of exposure, the number of motile ciliated cells significantly decreased. Direct measurement of PKCε activity revealed a dose-dependent activation of PKCε in response to NTHi supernatant exposure. Both CBF and PKCε activity changes were only observed in fresh NTHi culture supernatant and not observed in exposures to heat-inactivated or frozen supernatants.
Our results suggest that CBF slowing observed in response to NTHi is consistent with the stimulated activation of PKCε. Ciliated cell detachment is associated with PKCε autodownregulation.
鼻上皮的流感嗜血杆菌感染一直与观察到的鼻纤毛摆动频率（CBF）降低和纤毛上皮损伤有关。以前，我们曾报道过几种减缓 CBF 的药物也具有激活气道上皮细胞蛋白激酶 C ɛ（PKCε）活性的作用。随后 PKCε 的自动下调或 PKCε 的直接抑制导致纤毛细胞的特异性脱落。
原代培养的纤毛牛支气管上皮细胞暴露于过滤的无定型流感嗜血杆菌（NTHi）培养物的条件培养基上清液中。测量 CBF 和运动点，并测定 PKCε 活性。
NTHi 上清液暴露在 10 分钟内以剂量依赖性方式显著且快速降低 CBF。暴露 3 小时后，运动的纤毛细胞数量明显减少。直接测量 PKCε 活性显示 PKCε 在暴露于 NTHi 上清液时呈剂量依赖性激活。仅在新鲜的 NTHi 培养物上清液中观察到 CBF 和 PKCε 活性的变化，而在热失活或冷冻上清液中未观察到变化。
我们的结果表明，NTHi 引起的 CBF 减慢与 PKCε 的刺激激活一致。纤毛细胞脱落与 PKCε 的自动下调有关。
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