Investigation On The Objective Differences Of Cold And Hot Natural Property In Zuojin Wan And Its Similar Formulae Based On Biothermodynamics

The famous Chinese medicine formula-Zuojin Wan and similar formulae (Zuojin Wan, Ganlu San, Zhuyu Wan and Fanzuojin Wan), composed of Coptis and Evodia in different compatibility proportions, have totally differences not only in therapeutic features, but also obvious in their Cold and Hot natural properties. It has always been the hot and difficult spots in the study of Cold and Hot natural properties in Traditional Chinese Medicine. At present, the Cold and Hot natural properties shown by Chinese medicinal formulae has attracted much focus from the field. Under the guidance of Traditional Chinese Medicine and the Biothermodynamics theory, this paper illustrates the differences of two natural property and corresponding therapeutical effects of Zuojin Wan and its similar formulae on normal and Cold/Hot symptomatic mice by using the cold/hot plate differentiating technology and microcalorimetry combined with material basis analysis of chemical components. Furthermore, an objective method based on microcalorimetry was established to validate and estimate the authenticity and variability of Cold and Hot natural properties. The main research results are as follows:1. Study of animal temperature tropism praxiology:To study the effects of Zuojin Wan and its similar formulae on normal and Cold/Hot symptomatic mice based on mice temperature tropism, together with the authenticity and variability between Cold and Hot natural properties by a Cold/Hot plate differentiating technology. The results in the normal mice showed that the remaining ratio on warm pad would decreased with the decreasing of Coptis ratio in Zuojin Wan and its similar formulae. The order from high to low of the remaining ratio on warm pad was Coptis> Zuojinwan> Ganlu San≈Zhuyu Wan> Fanzuojin Wan> Evodia. On the other hand, in the Cold/Hot symptomatic mice, the results showed that the cold natural Zuojin Wan could decreased significantly the remaining rate of hot-symptom mice on warm pad (P<0.05), while was not notable to cold-symptom mice. The interference effects of Fanzuojin Wan on cold/hot symptom mice in cold-hot temperature tropism was adverse compared with Zuojin Wan. There was not significant difference and regular for Ganlu San and Zhuyu Wan to Cold/Hot symptomatic mice on the remaining rate of warm pad. Meanwhile, biochemical indicators which is relative to energy metabolism such as ATPase enzyme activity, total anti-oxidant capability (T-AOC) and superoxide dismutase (SOD) had corresponding change in organism.2. Study of Biothermodynamics:To investigate the effects of Zuojin Wan and its similar formulae on the thermogenic curves and corresponding thermodynamic parameter of the gastric cells of normal mice and Cold/Hot symptomatic mice by using principal component analysis and microcalorimetry. On normal gastric cells, the results showed that the order of growth and metabolism of normal gastric cells affected by the similar formulae was Fanzuojin Wan>Zhuyu Wan>Ganlu San>Zuojin Wan. Principal component analysis results indicated that 3 major parameters, namely the growth rate constant (κ), the maximum power output (Pm) and heat total power (Qt), represented the main information of effects of Zuojin wan and its similar formulae on growth and metabolism of the gastric cell. On Cold/Hot gastric cells, results showed that calorific value of growth and metabolism of the gastric cells was noticablely increased by Fanzuojin Wan, and the value of the Hot gastric cells was significantly decreased by Zuojin Wan. Compared with other similar formulae, their thermogenic curves were more similar with control. The principal component analysis result indicated that differences among the similar formulae decided by the thermodynamic parameters of both the regression coefficient (R) and the maximum power output (Pm) can considerably represent the main information with Zuojinwan and its similar formulae to the thermogenic curves of gastric cell of Cold/Hot symptomatic mice.3. Study of material basis of chemical components:to establish the UPLC fingerprints of Zuojin Wan and its similar formulae. Meanwhile, the content of eight main chemical constituents (alkaloids) were determined, the result of which was analyzed by the relative area normalization, similarity, cluster analysis and principal component analysis. Cluster analysis results indicated the objective differentiation Cold and Hot exists in various similar formulae of transformation. Both the relative normalized area of the fingerprint and the content from Zuojin Wan and its similar formulae showed that the change of the major chemical compositions (especially alkaloids) were the material base induced transformation between Cold and Hot. Finally, it was infered that material basis which affected the transformation between the Cold and Hot was X2, X3, X4, Xg, epiberberine hydrochloride, jateorrhizine hydrochloride, coptisine sulphate, palmatine hydrochloride and berberine hydrochloride by the chemical composition of the principal component analysis.In summary, the differences in Cold and Hot natural properties of Zuojin Wan and its similar formulae, composed of the same elements in different proportions, might be expressed by the external behavior of thermotropism and cellular level in an intuitive and objective way by using the Cold/Hot plate differentiating technology and microcalorimetry, verifying the scientific effectiveness of Chinese treatment principle and method of "expelling heat with cold herbs and cryopathy requiring warm prescription". Meanwhile, combining the the method of chemical component analysis, we have preliminarily determined the materals basis, which might influence the Cold and Hot natural properties of Zuojin Wan and its similar formulae. This research could provided a new visual angle and experimental reference for the study on the modernization of theory of Chinese Cold and Hot Natural Properties.