Studies on the effect of tetrandrine on microtubules. II. Observation with immunofluorescence techniques

Microtubules were purified by using two cycles of assembly and disassembly processes on fresh brain homogenates from 30 guinea pigs. The yield was about 60 mg. The effect of tetrandrine on tubulin was determined by spectrophotometric analysis and electron microscopy. In addition, we used the indirect immunofluorescent method including tubulin antibody to locate the presence of microtubules in 3T3 cells by fluorescence microscopy. The effects of colchicine and P204 were studied for comparison at the same time. The results showed that colchicine can effectively depolymerize microtubules, while tetrandrine showed aggregation, and in a different manner. The shape and structure of microtubules were definitely destroyed by colchicine, but were not affected by P204 which protected against the destructive effect of tetrandrine. This result indicated the safety of using a combination of P204 and tetrandrine in the treatment of silicosis.