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目的分析重症肌无力患者(MG)骨骼肌中碳酸酐酶Ⅲ(CAⅢ)蛋白和 CAⅢmRNA 的表达,并与健康对照组进行比较,探讨 MG 骨骼肌 CAⅢ蛋白缺乏的原因。方法用 Western blot 分析17例 MG 患者与19名健康人骨骼肌 CAⅢ蛋白的水平,逆转录-聚合酶链反应(RT-PCR)技术分析CAⅢ mRNA 的表达。结果 Western blot 显示,MG 患者骨骼肌 CAⅢ蛋白谱带密度低于健康人骨骼肌;经半定量分析,健康人骨骼肌 CAⅢ蛋白水平的相对值为1.70±0.29,MG 骨骼肌为0.76±0.08,两者差异有统计学意义(P=0.006)。RT-PCR 半定量结果为:健康人骨骼肌 CAⅢ mRNA 表达的相对值为0.29±0.04,MG 骨骼肌为0.15±0.02,两组间差异亦有统计学意义(P=0.005)。分析同一标本 CAⅢ蛋白与 CAⅢ mRNA 表达,约77%的 MG 患者骨骼肌中两者呈一致性减少。结论 MG 骨骼肌 CAⅢ蛋白降低的主要原因是其本身 CAⅢ mRNA 的表达降低所致,骨骼肌 CAⅢ mRNA 的表达降低与 MG 发病的关系还需进一步阐明。
Objective To analyze the expression of carbonic anhydrase Ⅲ (CA Ⅲ) protein and CA Ⅲ mRNA in skeletal muscle of patients with myasthenia gravis (MG) and to compare with the healthy control group to explore the reason of CA Ⅲ protein deficiency in MG skeletal muscle. Methods Western blot was used to detect the level of CA Ⅲ protein in skeletal muscle of 17 patients with MG and 19 healthy people. The expression of CA Ⅲ mRNA was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Results Western blot showed that the density of CA Ⅲ protein in skeletal muscle of MG patients was lower than that in healthy people. The relative value of CA Ⅲ protein in healthy people was 1.70 ± 0.29 and that of MG skeletal muscle was 0.76 ± 0.08 by semi-quantitative analysis The difference was statistically significant (P = 0.006). Semi-quantitative RT-PCR results showed that the relative value of CAⅢ mRNA expression in healthy skeletal muscle was 0.29 ± 0.04, and MG skeletal muscle was 0.15 ± 0.02. There was also a significant difference between the two groups (P = 0.005). Analysis of the same specimen CA Ⅲ protein and CA Ⅲ mRNA expression, about 77% of MG patients with skeletal muscle was consistent reduction. Conclusions The main reason for the decrease of CA Ⅲ protein in MG skeletal muscle is the decrease of its CA Ⅲ mRNA expression. The relationship between the decrease of CA Ⅲ mRNA in skeletal muscle and the pathogenesis of MG needs to be further elucidated.