目的通过观察按法干预对激痛点骨骼肌细胞骨架α微管蛋白(α-tubulin)和微管相关蛋白4(microtubule-associated protein 4,MAP-4)的影响,探讨按法的舒筋解结作用。方法40只SPF级雄性大鼠随机分为空白组10只和激痛点造模大鼠30只,采用钝...目的通过观察按法干预对激痛点骨骼肌细胞骨架α微管蛋白(α-tubulin)和微管相关蛋白4(microtubule-associated protein 4,MAP-4)的影响,探讨按法的舒筋解结作用。方法40只SPF级雄性大鼠随机分为空白组10只和激痛点造模大鼠30只,采用钝性打击结合离心运动的方法建立大鼠激痛点模型,模型评价后将符合标准的20只大鼠随机分为模型组和按法组,每组10只。空白组和模型组不予按法干预,仅正常观察,按法组以自制按法刺激器干预14 d。干预结束后在激痛点局部取材,用Western blot和免疫荧光双染法检测α-tubulin和MAP-4。结果α-tubulin和MAP-4在肌细胞和细胞外基质均有一定的共表达,空白组α-tubulin和MAP-4主要分布在肌细胞膜与细胞质,分布连续性较好,模型组α-tubulin分布连续性和完整性欠佳,在肌细胞内表达减少,在细胞外基质表达增加,MAP-4在肌细胞和细胞外基质表达均增加,两者在按法组有一定改善。与空白组相比,模型组α-tubulin表达下降、MAP-4表达升高(P<0.05);与模型组相比,按法组α-tubulin表达升高、MAP-4表达下降(P<0.05)。结论按法的舒筋解结作用可能与抑制微管蛋白解聚并促进其合成有关。展开更多
Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that af...Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that affect the microtubule (MT) stabilization, and the recruitment and activation of coat and cargo proteins, including ADP-ribosylation factors (Arfs) and protein kinase D (PKD). Here, we report on the factors implicated in the regulation of MMP-9 secretion by salivary gland acinar cells in response to P. gingivalis LPS, and the effect of hormone, ghrelin. We show that the LPS-elicited induction in MMP-9 secretion is associated with the increase in α-tubulin acetylation and the enhancement in MT stabilization, while the modulatory effect of ghrelin is reflected in a decrease in α-tubulin acetylation. Further, the effect of the LPS occurs in concert with up-regulation in Arf-guanine nucleotide exchange factor (GEF)-mediated Arf1 activation and the TGN recruitment of PKD2, while ghrelin exerts the modulatory effect on Arf-GEF activation. Moreover, we reveal that the LPS-induced up-regulation in MMP-9 secretion is reflected in a marked increase in PKCδ-mediated PKD2 phosphorylation on Ser, while the modulatory effect of ghrelin is manifested by the SFK-PTKs-dependent phosphorylation of PKD2 on Tyr. The findings demonstrate that MT stabilization along with Arf-GEF-mediated Arf1/PKD2 activation play a major role in P. gingivalis LPS-induced up-regulation in salivary gland acinar cell MMP-9 secretion, and point the modulatory mode of action by ghrelin.展开更多
文摘目的通过观察按法干预对激痛点骨骼肌细胞骨架α微管蛋白(α-tubulin)和微管相关蛋白4(microtubule-associated protein 4,MAP-4)的影响,探讨按法的舒筋解结作用。方法40只SPF级雄性大鼠随机分为空白组10只和激痛点造模大鼠30只,采用钝性打击结合离心运动的方法建立大鼠激痛点模型,模型评价后将符合标准的20只大鼠随机分为模型组和按法组,每组10只。空白组和模型组不予按法干预,仅正常观察,按法组以自制按法刺激器干预14 d。干预结束后在激痛点局部取材,用Western blot和免疫荧光双染法检测α-tubulin和MAP-4。结果α-tubulin和MAP-4在肌细胞和细胞外基质均有一定的共表达,空白组α-tubulin和MAP-4主要分布在肌细胞膜与细胞质,分布连续性较好,模型组α-tubulin分布连续性和完整性欠佳,在肌细胞内表达减少,在细胞外基质表达增加,MAP-4在肌细胞和细胞外基质表达均增加,两者在按法组有一定改善。与空白组相比,模型组α-tubulin表达下降、MAP-4表达升高(P<0.05);与模型组相比,按法组α-tubulin表达升高、MAP-4表达下降(P<0.05)。结论按法的舒筋解结作用可能与抑制微管蛋白解聚并促进其合成有关。
文摘Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that affect the microtubule (MT) stabilization, and the recruitment and activation of coat and cargo proteins, including ADP-ribosylation factors (Arfs) and protein kinase D (PKD). Here, we report on the factors implicated in the regulation of MMP-9 secretion by salivary gland acinar cells in response to P. gingivalis LPS, and the effect of hormone, ghrelin. We show that the LPS-elicited induction in MMP-9 secretion is associated with the increase in α-tubulin acetylation and the enhancement in MT stabilization, while the modulatory effect of ghrelin is reflected in a decrease in α-tubulin acetylation. Further, the effect of the LPS occurs in concert with up-regulation in Arf-guanine nucleotide exchange factor (GEF)-mediated Arf1 activation and the TGN recruitment of PKD2, while ghrelin exerts the modulatory effect on Arf-GEF activation. Moreover, we reveal that the LPS-induced up-regulation in MMP-9 secretion is reflected in a marked increase in PKCδ-mediated PKD2 phosphorylation on Ser, while the modulatory effect of ghrelin is manifested by the SFK-PTKs-dependent phosphorylation of PKD2 on Tyr. The findings demonstrate that MT stabilization along with Arf-GEF-mediated Arf1/PKD2 activation play a major role in P. gingivalis LPS-induced up-regulation in salivary gland acinar cell MMP-9 secretion, and point the modulatory mode of action by ghrelin.
