北京生物醫(yī)學(xué)工程

放散式體外沖擊波對軟骨細(xì)胞生物學(xué)行為的影響

Effect of radial extracorporeal shock wave on the biological behaviors of chondrocytes

作者：綦惠杰永生鄭蕊舒雄郭安憶王山劉亞軍

單位：北京市創(chuàng)傷骨科研究所（北京100035）北京積水潭醫(yī)院骨科沖擊波診療中心（北京100035）

關(guān)鍵詞：放散式體外沖擊波；骨關(guān)節(jié)炎；軟骨細(xì)胞；一氧化氮；生物學(xué)行為

分類號：   R318.04

出版年·卷·期（頁碼）：2020·39·3（278-284）

摘要：

目的放散式體外沖擊波（radial extracorporeal shock wave，rESW）能夠促進(jìn)軟骨缺損和骨關(guān)節(jié)炎（osteoarthritis，OA）軟骨損傷的修復(fù)，但具體機(jī)制仍需進(jìn)一步探討。本研究以軟骨細(xì)胞為研究對象，分析rESW對軟骨細(xì)胞生物學(xué)行為的影響及相關(guān)機(jī)制，為rESW的臨床應(yīng)用奠定研究基礎(chǔ)。方法采用膠原酶聯(lián)合胰蛋白酶消化的方法獲取大鼠軟骨細(xì)胞，通過甲苯胺藍(lán)染色對軟骨細(xì)胞進(jìn)行鑒定。分別在正常培養(yǎng)條件以及IL-1b介導(dǎo)的炎性條件下，給予不同能量的rESW刺激，采用CCK8法和劃痕法分別檢測軟骨細(xì)胞的增殖和遷移。通過對軟骨細(xì)胞培養(yǎng)上清中一氧化氮（nitric oxide，NO）的Griess法測定，檢測rESW對軟骨細(xì)胞NO分泌的影響。Western blot分析軟骨細(xì)胞誘導(dǎo)型一氧化氮合酶（inducible nitric oxide synthase，iNOS）的蛋白表達(dá)。結(jié)果正常條件下，rESW對軟骨細(xì)胞的增殖、遷移和NO分泌均未有顯著影響。IL-1b存在的炎性環(huán)境下，軟骨細(xì)胞增殖和遷移受到顯著抑制，rESW應(yīng)用后，軟骨細(xì)胞增殖能力有明顯恢復(fù)，但遷移未受影響。炎性環(huán)境下，軟骨細(xì)胞NO生成顯著升高，且iNOS蛋白表達(dá)上調(diào)，rESW抑制NO的分泌和iNOS蛋白的表達(dá)。結(jié)論炎性環(huán)境下，rESW能夠保護(hù)軟骨細(xì)胞的增殖功能，且抑制軟骨細(xì)胞分泌前炎癥因子NO，改善關(guān)節(jié)腔微環(huán)境，因此rESW能夠作為一種輔助手段應(yīng)用于關(guān)節(jié)軟骨損傷的治療。

Objective Radial extracorporeal shock wave (rESW) has curative effect on osteochondral lesions and osteoarthritis (OA). In order to find out the mechanisms of this effect, we apply rESW on chondrocytes directly to observe the changes of their biological behaviors under normal and inflammatory conditions. Methods Chondrocytes from cartilage of SD rats were achieved by enzymatic method. The cells were treated with interleukin 1 beta (IL-1β) as an in vitro inflammatory model. Three different energy levels of rESW were tested for the effects on the proliferation and migration of chondrocytes, with CCK8 method and cell scratch-wound assay, respectively. Nitric oxide (NO) secretion was detected through Griess method. Western blot was used to determine the protein expression of iNOS. Results The proliferation, migration and NO secretion were not changed under normal condition by rESW. When IL-1b was added, the proliferation of chondrocytes decreased, but recovered significantly after cells were treated with rESW; while the migration was not significantly affected by rESW. RESW significantly downregulated the NO level and inducible nitric oxide synthase (iNOS) protein expression in the presence of IL-1b. Conclusion RESW helps to protect the biological behaviors of chondrocytes under inflammatory condition, and may be used as a supplementary approach clinically in OA treatment.

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