Studies On The Method Of Evaluating And Screening Pi-Deficiency Therapy Traditional Chinese Medicine(TCM) By Guideline Of Intestinal Microflora

As one of commonly encountered diseases in clinic of Traditional Chinese Medicine (TCM), Pi-deficiency is one of the main syndromes of digestive system diseases and associated with intestinal microflora. In this research, a new evaluating and screening Pi-deficiency therapy TCM method was established by 16S rDNA technology.Rats were applied the decoction of Radix et Rhizoma Rhei to make pi-deficiency model, cured by Live Bacillus Licheniformis Craniles(oral) and the decoction of Ban-xia-hou-pu-tang, Si-jun-zi-tang, Li-zhong-tang, Bu-zhong-yi-qi-tang and Yi-wei-tang, the rat feces were collected in the process of essay. An inexpensive, efficient and reliable method for genomic DNA isolation of intestinal flora in fecal sample was established. A protocol of 16S rDNA Restriction Fragment Length Polymorphism (16S rDNA-RFLP) for fecal intestinal flora was optimized, two restriction endonucleases were selected based on the restriction endonucleases screening. The 16S rDNA-RFLP fingerprint of fecal intestinal microflora of healthy and pi-deficiency Rat were also established, and the characteristics of 16S rDNA-RFLP fingerprints of pi-deficiency rats was found by comparing with that of healthy. The treated rat intestinal microflora was also analysed, as a result, the evaluative systerm of Pi-deficiency therapy TCM by 16S rDNA-RFLP of intestinal flora was found. The curative effect of Pi-deficiency therapy TCM, Si-jun-zi-tang, Li-zhong-tang, Bu-zhong-yi-qi-tang and Yi-wei-tang, were also evaluated. These results showed the method established in this research could be used to evaluate the Pi-deficiency therapy TCM by comparing the 16S rDNA-RFLP fingerprints of intestinal microflora in different conditions. 16S rDNA-RFLP fingerprints reveals the whole intestinal microflora, has advantages compared to traditional methods such as culture and count.It’s the first reported that 16S rDNA-RFLP were used in TCM evaluating and screening. It also has important meaning to quantitative research of Pi-deficiency syndrome and the modernization of traditional Chinese theories.