Microcalorimetry coupled with statistical analysis techniques for bio-activity evaluation of medicinal animal horns and shells

Abstract

Microcalorimetry is an important analytical tool continuously measuring the heat output generated in the physical, chemical or biological processes. Here, the microcalorimetric method was used to evaluate the bio-activity differences of seven medicinal animal horns and shells on splenic lymphocytes. The cells were placed in the 20-mL glass ampoules containing RPMI-1640 and the test drugs (10 mg mL−1), and the heat output and motility of the cells were examined with an isothermal microcalorimeter. From the measured power–time curves, some quantitative thermokinetic parameters were obtained. By analyzing these parameters using principal component analysis, two main parameters, the maximum heat-output power P m and total heat output Q, were optimized, through which the effects of the seven drugs on splenic lymphocytes were quickly evaluated: 10 mg mL−1 of Cornu Cervi Pantotrichum showed stronger promotion effect than Cornu Cervi on the growth and proliferation of splenic lymphocytes, whereas the other five horns and shells animal drugs including Cornu Saigae Tataricae, Cornu Caprae Hircus, Cornu Bubali, Squama Manis and Carapax Trionycis at 10 mg mL−1 all inhibited the growth of splenic lymphocytes, and Cornu Saigae Tataricae expressed the strongest inhibitory effect. The result of hierarchical clustering analysis on the quantitative parameters indicated that the drugs with promotion effect on splenic lymphocytes clustered into one group and the other drugs with inhibitory effect consisted of another group. The results have highlighted the bio-activity differences of the seven horns and shells animal drugs referring to immune functions from the cell level, providing meaningful references for the clinical use of the valuable drugs.