| In this study, Houttuynia cordata thunb was used as the objective to discuss the quality stability from the perspective of genetic diversity among herbal plants. On the basis of the original GAP control mode, a mathematical model and a theoretical system "the minimum quantity for one feeding"(MQF) were established, which would attribute to further controlling the quality stability of Chinese herbal medicine (CHM). Firstly, a phenomenon that CHM had genetic polymorphism was revealed from the perspective of the chemical composition (fingerprints of CHM), gene and enzyme (genes and total proteins of CHM). Afterwards, we applied Hardy-Weinberg balance population law to study the individual of CHM with gene polymorphism. After combining the amount of information of CHM fingerprints, the results of ISSR and RAPD analysis, the mathematical model of MQF that could represent the Hardy-Weinberg balance population were established. Finally, after a large number of experimental researches on Houttuynia cordata thunb, we calculated the values of MQF by the mathematical model. Then the theory of MQF was established ultimately, which could provide a new ideas, new methods and experimental data for the quality control of the stability of CHM. The experimental results in this paper were listed as follows:1. The results for genetic diversity among the Houttuynia plants:the46plants of Houttuynia cordata from the same GAP base were analyzed by ISSR and RAPD, At the same time, cluster analysis was conducted by NTSYS2PC software. the results showed that the9ISSR primers and8RAPD primers were obtained after screening the134and101bands where the polymorphic number was115and72respectively, polymorphic ratio was85.8%,71.3%, the average genetic similarity coefficients were0.6786,0.6947. Two kinds of molecular marker results revealed the same GAP producing strains gene polymorphism of Houttuynia cordata. 1.2. The study on the proteins diversity of different plants of Houttuynia cordata:the Bradford method improved from Ramagli, SDS-PAGE gel electrophoresis and dual wavelength TLC scanning method were used in this study. It turned out that the molecular weights of protein bands from53plants were mostly concentrated range from6.5KD to97.2KD, the diversity of protein species was not obvious, but their colors varied apparently; the RSD of zero moment (AUCT), first moment (MCRTT) and second moment (VCRTT) of protein electrophoresis bands scan were40.92%,6.01%and18.57%respectively. Thus, the types of proteins from individual fresh Houttuynia cordata changed little under the same GAP, but the contents varied extremely.3. The results of the fingerprint diversity of volatile oil from Houttuynia plants:the volatile oil was extracted from77plants of Houttuynia cordata,77gas chromatographic fingerprints of volatile oil components were established, analyzed the fingerprints with the total statistical moments, the results showed that the RSD of total zero moment AUCT of volatile oil fingerprint from77plants Houttuynia cordata was341.56%, the RSD of total first moment MCRTTwas51.43%, the RSD of total second moment VCRTT was65.71%; therefore, the gas chromatography fingerprint of volatile oil between Houttuynia that cropped in the same GAP has great variability.4. The equilibrium population study based on the information entropy brought by the RAPD amplified bands of Houttuynia cordata: Using RAPD molecular markers to do polymorphism analysis on the same GAP origin Houttuynia cordata, and turned the information of amplified bands into the information entropy, then used mathematical model to measure the value of MQF of "Hardy-Weinberg" balance group of Houttuynia cordata. The results showed that101of total bands were obtained, the information entropy was H=0.4488～0.9033, RSD is 10.43%, and the number of "Hardy-Weinberg" equilibrium population was287strains, converting into MQF was about3.736kg.5. The equilibrium population study based on the information entropy brought by the ISSR amplified bands of Houttuynia cordata: Using ISSR molecular markers to do polymorphism analysis on the same GAP origin Houttuynia cordata, and turned the information of amplified bands into the information entropy, then used mathematical model to measure the value of MQF of "Hardy-Weinberg" balance group of Houttuynia cordata. The results showed that134of total bands were gotten, the information entropy was H=0.3656～0.9786, RSD was14.75%, and the number of "Hardy-Weinberg" equilibrium population was863strains, converting into MQF was about11.22kg.6. The equilibrium population study based on the information entropy brought by the gas chromatographic fingerprints of volatile oil of Houttuynia cordata:using information entropy and genetic statistical principle, turned the information of fingerprints into the information entropy, establishing mathematical model of CHM for MQF, meanwhile carrying on the experiment and the results showed that the number of genetic equilibrium population was29683strains, converted into MQF was about118.7kg.Conclusion:This paper revealed that the genetic diversity laws do exist among Houttuynia from fingerprinting variability, the difference of total protein and genetic polymorphisms by RAPD and ISSR. Using the information entropy theory and the statistics principles the relevant mathematical models were established, the calculated values of the three analytical methods were287strains,863strains and29683strains. The results showed that the values of the three different regions’equilibrium population were of great difference, there are two reasons:one is the internal cause, different regions, different environmental may lead the equilibrium population numerical to be different; the others are the different estimates angles, different expression forms of CHM’s polymorphism. Different forms would cause different value of equilibrium population. At last we should go through experimental verification and clinical efficacy testing, only in this way can we truly achieve the quality control theory of MQF for CHM, and make MQF theory more theoretical, feasibility and scientific. |
PDF Full Text Request