Background: Blocking the Rho A/ROCK Ⅱ/MLC 2(Ras homolog gene family member A/Rho kinase Ⅱ/myosin light chain 2) signaling pathway can initiate neuroprotective mechanisms against neurological diseases such as stroke,...Background: Blocking the Rho A/ROCK Ⅱ/MLC 2(Ras homolog gene family member A/Rho kinase Ⅱ/myosin light chain 2) signaling pathway can initiate neuroprotective mechanisms against neurological diseases such as stroke, cerebral ischemia, and subarachnoid hemorrhage. Nevertheless, it is not clear whether and how disrupting the Rho A/ROCK Ⅱ/MLC 2 signaling pathway changes the pathogenic processes of the blood–brain barrier(BBB) after intracerebral hemorrhage(ICH). The present investigation included the injection of rat caudal vein blood into the basal ganglia area to replicate the pathophysiological conditions caused by ICH. Methods: Scalp acupuncture(SA) therapy was performed on rats with ICH at the acupuncture point “Baihui”-penetrating “Qubin,” and the ROCK selective inhibitor fasudil was used as a positive control to evaluate the inhibitory effect of acupuncture on the Rho A/ROCK Ⅱ/MLC 2 signaling pathway. Post-assessments included neurological deficits, brain edema, Evans blue extravasation, Western blot, quantitative polymerase chain reaction, and transmission electron microscope imaging. Results: We found that ROCK Ⅱ acts as a promoter of the Rho A/ROCK Ⅱ/MLC 2 signaling pathway, and its expression increased at 6 h after ICH, peaked at 3 days, and then decreased at 7 days after ICH, but was still higher than the preintervention level. According to some experimental results, although 3 days is the peak, 7 days is the best time point for acupuncture treatment. Starting from 6 h after ICH, the neurovascular structure and endothelial cell morphology around the hematoma began to change. Based on the changes in the promoter ROCK Ⅱ, a 7-day time point was selected as the breakthrough point for treating ICH model rats in the main experiment. The results of this experiment showed that both SA at “Baihui”-penetrating “Qubin” and treatment with fasudil could improve the expression of endothelial-related proteins by inhibiting the Rho A/ROCK Ⅱ/MLC 2 signaling pathway and reduce neurological dysfunction, brain edema, and BBB permeability in rats. Conclusion: This study found that these experimental data indicated that SA at “Baihui”-penetrating “Qubin” could preserve BBB integrity and neurological function recovery after ICH by inhibiting Rho A/ROCK Ⅱ/MLC 2 signaling pathway activation and by regulating endothelial cell–related proteins.展开更多
Objective： To investigate the role of RhoA/ROCK in the process of chondrogenic differentiation of MSCs in vitro induced by TGF-β1. Methods ： MSCs were isolated from rat bone marrow, cultured and passaged. The cultu...Objective： To investigate the role of RhoA/ROCK in the process of chondrogenic differentiation of MSCs in vitro induced by TGF-β1. Methods ： MSCs were isolated from rat bone marrow, cultured and passaged. The cultured MSCs at the 3rd passage were induced to differentiate into chondrocytes in induction medium containing TGF-β1, the expressions of RhoA and ROCKI/2 in the induced MSCs cells were detected by Western-blot and RT-PCR. Results： In parallel to chondrogenic marker gene expressions of collagen Ⅱand aggrecan, ROCK inhibition by Y27632 in these cells caused a significant decrease in mRNA level of collagen Ⅱ. Conclusion： The results suggest that RhoA/ROCK pathway may play an important and complex role in regulation of chondrogenic differentiation and gene expression.展开更多
RhoA (Ras homolog gene family member A) belongs to the Rho subfamily of GTPases. ROCK (Rho—associated coiled—coil forming protein kinase) is downstream of the active RhoA and affects the generation and secretion of ...RhoA (Ras homolog gene family member A) belongs to the Rho subfamily of GTPases. ROCK (Rho—associated coiled—coil forming protein kinase) is downstream of the active RhoA and affects the generation and secretion of cellular element, which will result in relevant biologic effects. The RhoA/ROCK signaling pathway consists of these serious reactions. Therefore, the activation and inhibition of this pathway are closely related to the occurrence and development of many diseases. The research on the molecular mechanism of these diseases may be instructive and helpful to the clinical treatmen and prognosis of diseases. Recent studies of these typical diseases related to RhoA/ROCK signaling pathway are viewed in this article.展开更多
基金supported by the National Natural Science Foundation of China(numbers:81774416 and 81473764)。
文摘Background: Blocking the Rho A/ROCK Ⅱ/MLC 2(Ras homolog gene family member A/Rho kinase Ⅱ/myosin light chain 2) signaling pathway can initiate neuroprotective mechanisms against neurological diseases such as stroke, cerebral ischemia, and subarachnoid hemorrhage. Nevertheless, it is not clear whether and how disrupting the Rho A/ROCK Ⅱ/MLC 2 signaling pathway changes the pathogenic processes of the blood–brain barrier(BBB) after intracerebral hemorrhage(ICH). The present investigation included the injection of rat caudal vein blood into the basal ganglia area to replicate the pathophysiological conditions caused by ICH. Methods: Scalp acupuncture(SA) therapy was performed on rats with ICH at the acupuncture point “Baihui”-penetrating “Qubin,” and the ROCK selective inhibitor fasudil was used as a positive control to evaluate the inhibitory effect of acupuncture on the Rho A/ROCK Ⅱ/MLC 2 signaling pathway. Post-assessments included neurological deficits, brain edema, Evans blue extravasation, Western blot, quantitative polymerase chain reaction, and transmission electron microscope imaging. Results: We found that ROCK Ⅱ acts as a promoter of the Rho A/ROCK Ⅱ/MLC 2 signaling pathway, and its expression increased at 6 h after ICH, peaked at 3 days, and then decreased at 7 days after ICH, but was still higher than the preintervention level. According to some experimental results, although 3 days is the peak, 7 days is the best time point for acupuncture treatment. Starting from 6 h after ICH, the neurovascular structure and endothelial cell morphology around the hematoma began to change. Based on the changes in the promoter ROCK Ⅱ, a 7-day time point was selected as the breakthrough point for treating ICH model rats in the main experiment. The results of this experiment showed that both SA at “Baihui”-penetrating “Qubin” and treatment with fasudil could improve the expression of endothelial-related proteins by inhibiting the Rho A/ROCK Ⅱ/MLC 2 signaling pathway and reduce neurological dysfunction, brain edema, and BBB permeability in rats. Conclusion: This study found that these experimental data indicated that SA at “Baihui”-penetrating “Qubin” could preserve BBB integrity and neurological function recovery after ICH by inhibiting Rho A/ROCK Ⅱ/MLC 2 signaling pathway activation and by regulating endothelial cell–related proteins.
文摘目的 探究肝郁脾虚证的生物学基础及逍遥散的作用机制。方法 采用慢性束缚应激方法建立肝郁脾虚证大鼠模型。72只SD雄性大鼠随机分为6组,正常组,模型组,氟西汀组(0.0018 g/kg),逍遥散高(16.7 g/kg)、中(8.35 g/kg)、低(4.175g/kg)剂量组,每组12只。造模结束后,ELISA法测定大鼠血清单丝氨酸蛋白激酶1(single serine protein kinase 1,LIMK1)的含量,荧光定量PCR、 Western blot检测海马组织RAS同源基因家族成员A(RAS homologous gene family member A,RhoA)、Rho相关螺旋卷曲蛋白激酶2(Rho-related spiral coiled protein kinase 2,ROCK2)的m RNA及蛋白表达。结果 模型组大鼠血清LIMK1含量,海马组织中RhoA、ROCK2的mRNA和蛋白表达水平均明显高于正常组(P<0.01);各药物组大鼠血清LIMK1含量明显低于模型组(P<0.01),逍遥散高、中剂量组和氟西汀组大鼠海马组织中RhoA、ROCK2的mRNA和蛋白表达明显低于模型组(P<0.05,P<0.01)。结论 慢性束缚应激肝郁脾虚证大鼠海马组织RhoA/ROCK2通路被激活,逍遥散可抑制RhoA/ROCK2通路激活,并且作用呈剂量依赖性。
基金National Natural Science Foundation of China(No.30471753)
文摘Objective： To investigate the role of RhoA/ROCK in the process of chondrogenic differentiation of MSCs in vitro induced by TGF-β1. Methods ： MSCs were isolated from rat bone marrow, cultured and passaged. The cultured MSCs at the 3rd passage were induced to differentiate into chondrocytes in induction medium containing TGF-β1, the expressions of RhoA and ROCKI/2 in the induced MSCs cells were detected by Western-blot and RT-PCR. Results： In parallel to chondrogenic marker gene expressions of collagen Ⅱand aggrecan, ROCK inhibition by Y27632 in these cells caused a significant decrease in mRNA level of collagen Ⅱ. Conclusion： The results suggest that RhoA/ROCK pathway may play an important and complex role in regulation of chondrogenic differentiation and gene expression.
文摘RhoA (Ras homolog gene family member A) belongs to the Rho subfamily of GTPases. ROCK (Rho—associated coiled—coil forming protein kinase) is downstream of the active RhoA and affects the generation and secretion of cellular element, which will result in relevant biologic effects. The RhoA/ROCK signaling pathway consists of these serious reactions. Therefore, the activation and inhibition of this pathway are closely related to the occurrence and development of many diseases. The research on the molecular mechanism of these diseases may be instructive and helpful to the clinical treatmen and prognosis of diseases. Recent studies of these typical diseases related to RhoA/ROCK signaling pathway are viewed in this article.
