Effect of cigarette smoke extract on the role of protein kinase C in the proliferation of passively sensitized human airway smooth muscle cells / 华中科技大学学报（医学）（英德文版）

To investigate the effect of cigarette smoke extract (CSE) on the role of protein kinase C (PKC) in the proliferation of passively sensitized human airway smooth muscle cells (HASMCs). After synchronization of cultured HASMCs, they were divided into a group A and Group B. The group A was treated with normal human serum and served as controls and the group B was treated with the serum of asthma patients. The group A was further divided into group of A1, A2 and A3 and the group B was sub-divided into the group of B1, B2, B3, B4 and B5. No other agents were added to the group A1 and B1. The cells of group A2 and B2 were stimulated with 5% CSE for 24 h. HASMCs from group A3 and B3 were treated with PKC agonist PMA (10 nmol/L) and CSE (5%) for 24 h. PKC inhibitor Ro-31-8220 (5 micromol/L) was added to the HASMCs of group B4 for 24 h. The cells from group B5 were stimulated with Ro-31-8220 (5 micromol/L) and CSE (5 %) for 24 h. The proliferation of HASMCs isolated from group A and B was examined by cell cycle analysis, MTT colorimetric assay and 3H-TdR incorporation test. The expression of PKC-a in each group was observed by Western blotting and RT-PCR, respectively. The results showed that the percentage of S phase, absorbance (A) value, the rate of 3H-TdR incorporation, the ratios of A value of PKC-alpha mRNA and the A value of PKC-alpha protein in HASMCs from group B1, B2 and B3 were significantly increased compared to those of group A1, A2 and A3 correspondingly and respectively (P< 0.01). The proliferation of HASMCs of group A2 and B2 stimulated with CSE and group A3 and B3 stimulated with CSE and PMA were also significantly enhanced when group A1, A2 and A3 and group B1, B2 and B3 compared to each other (P<0.05, P<0.01, respectively). The percentage of S phase, absorbency (A) value, 3H-TdR incorporation rate, the ratios of A value of PKC-alpha mRNA and the A value of PKC-alpha protein in HASMCs from group B4 treated with Ro-31-8220 and group B5 treated with CSE and Ro-31-8220 were significantly decreased as compared to those of group B1 and B2 correspondingly and respectively (P<0.05, P<0.01). It was concluded that CSE can enhance the passively sensitized HASMC proliferation and the expression of PKC alpha. PKC and its alpha subtype may contribute to this process. Our results suggest cigarette may play an important role in ASMCs proliferation of asthma through PKC signal pathway.

Co-localization of c-fos protein and protein kinase C gamma in the rat brain following anodal polarization.

The expression of protein kinase c gamma (PKC gamma) and c-fos protein was examined by means of double labeling in the rat brain in relation to the molecular mechanism of central plastic changes associated with anodal polarization. Under normal, non-polarized condition, approximately 75% of all fos positive neurons in the neocortex were immunopositive for PKC gamma. Conversely, nearly all PKC gamma positive neurons were fos immunopositive. Although both pyramidal and non-pyramidal neurons express both types of protein, the pyramidal cell type represents the vast majority. An anodal direct current of 3.0 microA for 30 min to the surface of the left sensorimotor cortex resulted in a pronounced increase in the intensity of immunoreactivity for both PKC gamma and c-fos protein ipsilateral to the polarization. Approximately, 91% of fos positive neurons in the polarized neocortex was also intensely immunoreactive for PKY gamma. The high degree of codistribution of both transduction proteins in specific neurons following anodal polarization suggests the functional connection between PKY gamma activation and c-fos expression in polarization phenomenon.

Cambios en la distribucion y expresion eosenzimatica de proteina quinasa C en linfocitos B por estimulacion aloantigenica / Changes in distribution and isoenzimatic expression of protein kinase C in B cells by alloantigenic stimulation

La proteína qinasa C (PKC) comprende una familia de isoenzimas involucradas en la transducción de señales vía la estimulación de proteínas de membrana. Estas isoenzimas difieren en su distribución tisular, requerimientos de activación y especificidad de sustrato. Recientes estudios muestran la regulación en la expresión de isoformas de PKC, en diferentes situaciones fisiológicas y patológicas. En un trabajo previo demostramos la participación de la PKC en la respuesta de células B a células alogeneicas intactas y a aloantígeno solubilizado. En el presente estudio se analizó la influencia de la aloinmunización sobre la expresión de isoformas de PKC en células B, De acuerdo a nuestros resultados las células B expresan principalmente la isoforma ß, em menor proporción las Ó, õ y y en muy poca cantidad la Ù. El análisis realizado con linfocitos de animales previamente inmunizados indicó que a medida que aumenta el número de inmunizaciones, aumenta la expresión de las isoformas Ó y õ y disminuye la expresión de la ß. Además se observó una mayor proporción de enzima unida a membrana. Por otro lado, a medida que aumenta el número de proliferativa al lipopolisacárido (LPS). En cambio, la falta de respuesta proliferativa frente al aloantígeno solubilizado observada para células B sin inmunizar pasa a ser francamente proliferativa para células con 5-6 inmunizaciones. Estos resultados indican que la aloinmunización induce cambios en la expresión y distribución de las isoformas de la enzima que se correlacionan con cambios en la respuesta biológica obtenida por estimulación