目的研究体外人骨髓间充质干细胞(MSCs)和汗腺细胞(SGCs)直接和间接共培养条件下骨髓间充质干细胞的表型转化及其机制。方法体外分别分离培养、扩增并鉴定MSCs和汗腺细胞。将培养的MSCs和经47℃高温处理造成热休克的SGCs直接和间接共培...目的研究体外人骨髓间充质干细胞(MSCs)和汗腺细胞(SGCs)直接和间接共培养条件下骨髓间充质干细胞的表型转化及其机制。方法体外分别分离培养、扩增并鉴定MSCs和汗腺细胞。将培养的MSCs和经47℃高温处理造成热休克的SGCs直接和间接共培养,1周后,采用免疫细胞化学染色法和流式细胞仪法检测共培养体系中MSCs的表型改变,W estern b lot测定细胞外信号调节激酶(ERK)和磷酸化细胞外信号调节激酶(pERK)表达。结果MSCs和SGCs均呈克隆样生长,MSCs表达CD44、CD105和CD29,不表达CD34、CEA、CK19和CK7;SGCs表达CEA、CK19、CK8和CK7。MSCs与经高温损伤后的SGCs共培养1周后,部分MSCs呈汗腺细胞表型,间接共培养结果示各组MSCs均表达水平相当的ERK,但pERK水平表达不同。结论成人MSCs和热休克的SGCs直接和间接共培养均可诱导MSCs向SGCs表型转化,pERK途径参与这一过程。展开更多
目的探讨丙泊酚麻醉对胎鼠p Ca MKIIα/Ca MKIIα的影响。方法健康孕14 d左右SD大鼠63只,按照随机数字表法分为3组(n=21):对照组(C组)、单次丙泊酚组(PS组)和多次丙泊酚组(PM组)。PS组腹腔注射生理盐水7.5 ml/(kg·d),连续6 d,第7...目的探讨丙泊酚麻醉对胎鼠p Ca MKIIα/Ca MKIIα的影响。方法健康孕14 d左右SD大鼠63只,按照随机数字表法分为3组(n=21):对照组(C组)、单次丙泊酚组(PS组)和多次丙泊酚组(PM组)。PS组腹腔注射生理盐水7.5 ml/(kg·d),连续6 d,第7天注射丙泊酚75 mg/kg;PM组腹腔注射丙泊酚75 mg/(kg·d),连续7 d。第7天注射后2 h,测定血气和血糖(各组5只),其余孕鼠剖宫取胎鼠,采用免疫组织化学法和Western blot法检测胎鼠海马Ca MKIIα及p Ca MKIIα的表达。结果各组血气和血糖值均在正常范围内。与C组比较,PM组Ca MKIIα和p Ca MKIIα含量降低及p Ca MKIIα/Ca MKIIα比值下降(P<0.05),PS组各指标差异无统计学意义(P>0.05)。结论多次丙泊酚麻醉导致胎鼠脑内Ca MKIIα和p Ca MKIIα表达下调,使发育高峰期中枢神经系统受损。展开更多
AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immun...AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immunochemistry. Transcriptional and posttranscriptional levels of Ca MKⅡin tissue samples and MMP2,MMP9 and TIMP-1 expression in the human colon cancer cell line HCT116 were assessed by q RTPCR and western blot. Cell proliferation was detected with the MTT assay. Cancer cell migration and invasion were investigated with the Transwell culture system and woundhealing assay.RESULTS We first demonstrated that CaMK Ⅱ was ove rexpressed in human colon cancers and was associated with cancer differentiation. In the human colon cancer cell line HCT116,the Ca MKII-specific inhibitor KN93,but not its inactive analogue KN92,decreased cancer cell proliferation. Furthermore,KN93 also significantly prohibited HCT116 cell migration and invasion. The specific inhibition of ERK1/2 or p38 decreased the proliferation and migration of colon cancer cells.CONCLUSION Our findings highlight Ca MKⅡ as a potential critical mediator in human colon tumor development and metastasis.展开更多
In rice, the Ca^(2+)/calmodulin-dependent protein kinase OsDMI3 is an important positive regulator of abscisic acid (ABA) signaling. In ABA signaling, H_(2)O_(2) is required for ABA-induced activation of OsDMI3, which...In rice, the Ca^(2+)/calmodulin-dependent protein kinase OsDMI3 is an important positive regulator of abscisic acid (ABA) signaling. In ABA signaling, H_(2)O_(2) is required for ABA-induced activation of OsDMI3, which in turn increase H_(2)O_(2) production. However, how OsDMI3 regulates H_(2)O_(2) production in ABA signaling remains unknown. Here we show that OsRbohB is the main NADPH oxidase involved in ABA-induced H_(2)O_(2) production and ABA-mediated physiological responses. OsDMI3 directly interacts with and phosphorylates OsRbohB at Ser-191, which is OsDMI3-mediated site-specific phosphorylation in ABA signaling. Further analyses revealed that OsDMI3-mediated OsRbohB Ser-191 phosphorylation positively regulates the activity of NADPH oxidase and the production of H_(2)O_(2) in ABA signaling, thereby enhancing the sensitivity of seed germination and root growth to ABA and plant tolerance to water stress and oxidative stress. Moreover, we discovered that the OsDMI3-mediated OsRbohB phosphorylation and H_(2)O_(2) production is dependent on the sucrose non-fermenting 1-related protein kinases SAPK8/9/10, which phosphorylate OsRbohB at Ser-140 in ABA signaling. Taken together, these results not only reveal an important regulatory mechanism that directly activates Rboh for ABA-induced H_(2)O_(2) production but also uncover the importance of this regulatory mechanism in ABA signaling.展开更多
文摘目的研究体外人骨髓间充质干细胞(MSCs)和汗腺细胞(SGCs)直接和间接共培养条件下骨髓间充质干细胞的表型转化及其机制。方法体外分别分离培养、扩增并鉴定MSCs和汗腺细胞。将培养的MSCs和经47℃高温处理造成热休克的SGCs直接和间接共培养,1周后,采用免疫细胞化学染色法和流式细胞仪法检测共培养体系中MSCs的表型改变,W estern b lot测定细胞外信号调节激酶(ERK)和磷酸化细胞外信号调节激酶(pERK)表达。结果MSCs和SGCs均呈克隆样生长,MSCs表达CD44、CD105和CD29,不表达CD34、CEA、CK19和CK7;SGCs表达CEA、CK19、CK8和CK7。MSCs与经高温损伤后的SGCs共培养1周后,部分MSCs呈汗腺细胞表型,间接共培养结果示各组MSCs均表达水平相当的ERK,但pERK水平表达不同。结论成人MSCs和热休克的SGCs直接和间接共培养均可诱导MSCs向SGCs表型转化,pERK途径参与这一过程。
文摘目的探讨丙泊酚麻醉对胎鼠p Ca MKIIα/Ca MKIIα的影响。方法健康孕14 d左右SD大鼠63只,按照随机数字表法分为3组(n=21):对照组(C组)、单次丙泊酚组(PS组)和多次丙泊酚组(PM组)。PS组腹腔注射生理盐水7.5 ml/(kg·d),连续6 d,第7天注射丙泊酚75 mg/kg;PM组腹腔注射丙泊酚75 mg/(kg·d),连续7 d。第7天注射后2 h,测定血气和血糖(各组5只),其余孕鼠剖宫取胎鼠,采用免疫组织化学法和Western blot法检测胎鼠海马Ca MKIIα及p Ca MKIIα的表达。结果各组血气和血糖值均在正常范围内。与C组比较,PM组Ca MKIIα和p Ca MKIIα含量降低及p Ca MKIIα/Ca MKIIα比值下降(P<0.05),PS组各指标差异无统计学意义(P>0.05)。结论多次丙泊酚麻醉导致胎鼠脑内Ca MKIIα和p Ca MKIIα表达下调,使发育高峰期中枢神经系统受损。
基金Supported by the National Natural Science Foundation of China,No.81302131
文摘AIM To investigate the role of calmodulin-dependent protein kinase Ⅱ(Ca MKⅡ) in colon cancer growth,migration and invasion.METHODS Ca MKⅡ expression in colon cancer and paracancerous tissues was evaluated via immunochemistry. Transcriptional and posttranscriptional levels of Ca MKⅡin tissue samples and MMP2,MMP9 and TIMP-1 expression in the human colon cancer cell line HCT116 were assessed by q RTPCR and western blot. Cell proliferation was detected with the MTT assay. Cancer cell migration and invasion were investigated with the Transwell culture system and woundhealing assay.RESULTS We first demonstrated that CaMK Ⅱ was ove rexpressed in human colon cancers and was associated with cancer differentiation. In the human colon cancer cell line HCT116,the Ca MKII-specific inhibitor KN93,but not its inactive analogue KN92,decreased cancer cell proliferation. Furthermore,KN93 also significantly prohibited HCT116 cell migration and invasion. The specific inhibition of ERK1/2 or p38 decreased the proliferation and migration of colon cancer cells.CONCLUSION Our findings highlight Ca MKⅡ as a potential critical mediator in human colon tumor development and metastasis.
基金supported by the National Natural Science Foundation of China(31971824 and 32170316).
文摘In rice, the Ca^(2+)/calmodulin-dependent protein kinase OsDMI3 is an important positive regulator of abscisic acid (ABA) signaling. In ABA signaling, H_(2)O_(2) is required for ABA-induced activation of OsDMI3, which in turn increase H_(2)O_(2) production. However, how OsDMI3 regulates H_(2)O_(2) production in ABA signaling remains unknown. Here we show that OsRbohB is the main NADPH oxidase involved in ABA-induced H_(2)O_(2) production and ABA-mediated physiological responses. OsDMI3 directly interacts with and phosphorylates OsRbohB at Ser-191, which is OsDMI3-mediated site-specific phosphorylation in ABA signaling. Further analyses revealed that OsDMI3-mediated OsRbohB Ser-191 phosphorylation positively regulates the activity of NADPH oxidase and the production of H_(2)O_(2) in ABA signaling, thereby enhancing the sensitivity of seed germination and root growth to ABA and plant tolerance to water stress and oxidative stress. Moreover, we discovered that the OsDMI3-mediated OsRbohB phosphorylation and H_(2)O_(2) production is dependent on the sucrose non-fermenting 1-related protein kinases SAPK8/9/10, which phosphorylate OsRbohB at Ser-140 in ABA signaling. Taken together, these results not only reveal an important regulatory mechanism that directly activates Rboh for ABA-induced H_(2)O_(2) production but also uncover the importance of this regulatory mechanism in ABA signaling.
