ABSTRACT
Objective To explore the expression status of carbonic anhydrase Ⅲ (CA Ⅲ) from quadriceps femoris muscle in two kinds of muscle clinical phenotype (skeletal muscle atrophy group and skeletal muscles non-atrophy group) of chronic obstructive pulmonary disease (COPD).Methods Totally 37 inpatients from our hospital,were divided into 11 patients without COPD and 26 patients with COPD,in addition,according to body mass index,fat free mass index and quadriceps cross-section diameter,patients with COPD were divided into 14 skeletal muscles non-atrophy patients (SMNA)and 12 skeletal muscle atrophy patients (SMA).CA Ⅲ concentration of femoris quadriceps specimens was quantitatively determined using Western blot methods,CA Ⅲ mRNA expression levels of femoris quadriceps specimens were also quantitatively measured using RT-PCR,then compared among the 3 groups.Results There was significant difference in CA Ⅲ quantitative concentration and CA Ⅲ mRNA expression level in each group (P ＜ 0.05),further more,CA Ⅲ concentration expression level was significantly higher (P ＜ 0.01)in SMA group (1.260 ± 0.068) than in SMNA group (1.110 ± 0.014),the latter was significantly higher (P ＜ 0.01) than in the control group (1.000 ± O.062).CA Ⅲ mRNA expression level was significantly higher(P ＜ 0.01) in SMNA group(2.170 ±0.412) than in the control group(1.000 ±0.115),and was significantly lower than in SMA group (3.770 ± 0.788 ; P ＜ O.01).CA Ⅲ concentration and CA Ⅲ mRNA expression level increased at equal pace in SMNA group and SMA group,however,CA Ⅲ quantitative concentration and CA Ⅲ mRNA expression level were inconsistent in the two groups.Conclusion The expression status of CA Ⅲ in quadriceps femoris muscle was different in two kinds of muscle clinical phenotype of COPD.
ABSTRACT
Objective To observe the effects of nerve impulses on the expression of carbonic anhydrase Ⅲ ( CAⅢ ) and its phosphatase activity, and to explore whether or not the cause of CAⅢ expressive decreased in skeletal muscles of myasthenia gravis( MG) is resulted from the obstruction of nerve impulse.Methods The motor nerves of extensor digitorum longus (EDL, mainly composed by fast fibers) and soleus (Sol, mainly composed by slow fibers) were cut off by operation of denervation.Levels and phosphatase activities of CAⅢ were analyzed at 7, 14, 28, and 56 d after denervation by Western blot and specific enzyme staining on the membrane following SDS-polyacrylamide gel electrophoresis, respectively.Results (1) Levels of CAⅢ in Sol of normal side (eg denervated contralateral) were much higher than that in EDL of normal side, and the levels in both Sol and EDL had an enhanced tendency with time (age) increase, especially for Sol.After denervation, the levels of CAⅢ in EDL were gradual increased, however, the level in Sol was 14 d after denervation as the boundary of ascension and then decline.( 2) The phosphatase activities of CAⅢ in Sol of normal sides were much higher than that in EDL of normal sides, and there were an enhanced tendency with time (age) increase in Sol, but no significant changes were found in EDL The enzyme activities in denervated Sol were lower(in the 14, 28, and 56 days after denervation: 14.39 ±1.93, 11.48 ±1.46, 9.04 ±1.46) much than their contralaterals(22.75 ± 1.80, 25.26 ±3.15, 25.82 ± 2.97; t = 0.002, 0.005, 0.002, all P ＜ 0.05), the enzyme activities in denervated EDL were also lower than their contralaterals, however, no significant differences were found.(3)It was consistent for CAⅢ levels and phosphatase activities in both Sol and EDL of normal sides.After denervation, however, the deviation of the CAⅢ levels and phosphatase activities happened, the levels of CAⅢ were increased, but the phosphatase activities were decreased.Conclusions The effect of nerve impulse transferring obstructed by denervation on CAⅢ expression of skeletal muscles is different from that by MG auto-antibody.The decrease of CAⅢ protein in the MG muscles may be not resulted from the nerve impulse transferring obstructed by MG auto-antibody.
ABSTRACT
Objective To demonstrate the carbonic anhydrase Ⅲ (CAⅢ) for 25 000 protein decreased in skeletal muscle of myasthenia gravis (MG). Methods The protein molecular properties responsible to antibodies against 25 000 protein and CAⅢ were analyzed by a combination method of two-dimensional electrophoresis and immuno-Western blot. Competitive binding reactions of the antibodies to the purified 25 000 protein and muscular homogenate were observed by using immuno-Dot blot and immuno-Western blot, respectively. The expression of CAⅢ from normal and MG muscles was detected by immuno-Western blot. Results Combination analysis of two-dimensional electrophoresis and immuno-Western blot showed that the protein of immunological responsible to antibodies against 25 000 protein and CAⅢ had an identical molecular mass and isoelectric point. Competitive binding reactions proved that 25 000 protein and CAⅢ were the same substance, either by immuno-Dot blot or by immuno-Western blot. In addition, a much similar result was obtained when the levels of 25 000 protein from normal and MG muscles were detected by antibodies against 25 000 protein and (CAⅢ) by immuno-Western blot. Conclusion 25 000 protein decreased in the MG skeletal muscle was proved to be just a known protein CAⅢ, which made a basis for further exploring the relationship of CAⅢ deficiency and MG pathogenesis.