Meltzer Kate R, Standley Paul R
University of Arizona College of Medicine, 550 E Van Buren St, Room 3353, Phoenix, AZ 85004-2230, USA.
J Am Osteopath Assoc. 2007 Dec;107(12):527-36.
Clinical studies have supported the efficacy of a variety of osteopathic manipulative techniques. However, an evidence base for the cellular mechanisms underlying these clinical findings is lacking.
To investigate human fibroblast proliferation and interleukin secretory profiles in response to modeled repetitive motion strain (RMS) and modeled indirect osteopathic manipulative techniques (IOMT). The authors hypothesized that the RMS model would increase fibroblast proliferation and proinflammatory interleukin secretion, while the IOMT model would reverse these effects.
Human fibroblasts were exposed in vitro to one of three conditions: (1) an 8-hour RMS; (2) a 60-second IOMT; or (3) an 8-hour RMS followed by a 60-second IOMT. Data on fibroblast proliferation and interleukins present in conditioned media were obtained immediately after RMS, at 24 hours after RMS (24RMS), at 24 hours after IOMT (24IOMT), and at 24 hours after RMS and IOMT (24RMS+IOMT). Cytokine protein array and enzyme-linked immunosorbent assay were used in data analysis. Fibroblast proliferation was also measured colorimetrically with a cell proliferation assay.
Fibroblasts that underwent RMS secreted several proinflammatory interleukins 24 hours after strain cessation, with substantially increased secretion of IL-1alpha, IL-1beta, IL-2, IL-3, IL-6, and IL-16. At 24 hours after strain cessation, fibroblasts subjected to RMS also secreted increased amounts of the anti-inflammatory IL-1ra, and they displayed 15% less proliferation, compared with baseline cells (P<.05). Fibroblasts that underwent IOMT, when analyzed at 24 hours after IOMT, did not display increased interleukin secretion or proliferation. However, they did display a 44% reduction in proinflammatory IL-3 secretion when compared with baseline cells (P<.05). The use of 24RMS+IOMT did not induce interleukin secretion in fibroblasts that were analyzed 24 hours after the combined exposure. However, cells in the 24RMS+IOMT group did display a 46% reduction in proinflammatory IL-6 secretion compared with RMS alone (24RMS; P<.05), as well as a 51% increase in proliferation compared with the 24RMS group (P<.05).
An in vitro strain model that simulates RMS has different effects on fibroblast proliferation and interleukin secretion than does an in vitro model that simulates IOMT. Modeled RMS appears to cause a reduction in fibroblast proliferation and a delayed inflammatory response. Modeled IOMT not only fails to induce this response, it also reverses inflammatory effects in cells that have been strained repetitively. Data from the present study suggest that fibroblast proliferation and expression/secretion of proinflammatory and anti-inflammatory interleukins may contribute to the clinical efficacy of indirect osteopathic manipulative techniques.
临床研究已证实多种整骨手法技术的疗效。然而，这些临床发现背后的细胞机制缺乏证据支持。
研究人体成纤维细胞对模拟重复性运动应变（RMS）和模拟间接整骨手法技术（IOMT）的增殖及白细胞介素分泌情况。作者假设RMS模型会增加成纤维细胞增殖和促炎白细胞介素分泌，而IOMT模型会逆转这些效应。
将人体成纤维细胞在体外暴露于三种条件之一：（1）8小时的RMS；（2）60秒的IOMT；或（3）8小时的RMS后接着60秒的IOMT。在RMS后即刻、RMS后24小时（24RMS）、IOMT后24小时（24IOMT）以及RMS和IOMT后24小时（24RMS+IOMT）获取条件培养基中有关成纤维细胞增殖和白细胞介素的数据。数据分析采用细胞因子蛋白阵列和酶联免疫吸附测定。还用细胞增殖测定法通过比色法测量成纤维细胞增殖情况。
经历RMS的成纤维细胞在应变停止24小时后分泌多种促炎白细胞介素，IL-1α、IL-1β、IL-2、IL-3、IL-6和IL-16的分泌大幅增加。在应变停止24小时时，经历RMS的成纤维细胞还分泌了更多的抗炎性IL-1ra，与基线细胞相比，其增殖减少了15%（P<0.05）。在IOMT后24小时分析时，经历IOMT的成纤维细胞未显示白细胞介素分泌增加或增殖增加。然而，与基线细胞相比，它们的促炎IL-3分泌减少了44%（P<0.05）。联合使用24RMS+IOMT在联合暴露后24小时分析时未诱导成纤维细胞分泌白细胞介素。然而，与单独的RMS（24RMS）相比，24RMS+IOMT组的细胞促炎IL-6分泌减少了46%（P<0.05），与24RMS组相比增殖增加了51%（P<0.05）。
模拟RMS的体外应变模型与模拟IOMT的体外模型对成纤维细胞增殖和白细胞介素分泌的影响不同。模拟的RMS似乎会导致成纤维细胞增殖减少和炎症反应延迟。模拟的IOMT不仅未能诱导这种反应，还能逆转反复应变细胞中的炎症效应。本研究数据表明，成纤维细胞增殖以及促炎和抗炎白细胞介素的表达/分泌可能有助于间接整骨手法技术的临床疗效。
