A Preliminary Study On The Pathogenesis Underlying Nasopharyngeal Precancerous Lesion Induced By The Interactive Effects Of Qi-dificient Physique Status With N,N'-dinitrosopiperazine

Nasopharyngeal carcinoma (NPC) is the most commonly seen malignancy in the region of head and neck in southern China and Southeast Asia and closely related with precanceron lesion in the nasopharynx in its iniation and development. In order to investigate the possible role of Qi-dificient physique status in the pathogenesis underlying of nasopharyngeal precancerous lesions, the interactive effects of such a status of physique with environmental factor on this pathologic process were carried out in the following four aspects.Firstly, a survey was made to explore the distributing characteristic of the physique types in the terms of Traditional Chinese Medicine (TCM) in the population with various titers of EB virus associated antibody IgA/VCA. 150 cases were sampled from 784 persons consulting at the out-pationt clinic of Zhongshan TCM Hospital as the surveying object. Then, IgA/VCA titers in serum were determined and the types of physique status were investigated among them to analyze the distribution characteristics of the physique types in population with EBV infection and their association with EBV infecting activity as indicated by the level of IgA/VCA. The results showed that the rate of cases with a physique type of Qi-deficiency was significantly higher than that of other types of physique status (P<0.05) and normal physique status and imbalanced mixed with dampness were significant lower than other types (P<0.05) among this sample of population.Secondly, experiment was carried out to explore the delayed changing features in gene expressive profile of nasopharyngeal tissues among rats following Qi-deficiency modeling for a longer period. Male SD rats at the age of 4 weeks were modeled by exhausted swimming method in a deep pool, one time a day and lasted for 30 days, and the tissue samples of nasopharyngeal mucosa were taken 100 days late from them. Then, the total RNA was extracted from the tissue samples by hybridization was made with probe. Followed was fluorescence scanning and signal analysis to screen the differentially expressed gene in these samples. As a result of these procedures, 117 genes were detected as differentially expressed genes, with 41 up-regulated and 76 down regulated. Among those down regulated in their expressed activity, the most significance expressing genes were IL-1βand its related genes. Moreover, the expression of GstaW1 and GstaW2 genes were upregulated and that of Gsta synthesize enzyme gene was down regulated.Thirdly, studies were made to investigate the interactive effects of Qi-deficient physique status with N, N'-dinitrosopiperazine (DNP ) on the ultrastructural pathology of lesion of nasopharyngeal mucosa among rats with precancerom in the imitation and development of the nasopharyngeal mucosa, and the intervening effects of compound components extracted from astragalus and coptis on this lesion induced by such an intervention effect. Two hundred days late following exhausted swimming to induce Qi-deficiency and DNP injection to induce nasopharyngeal carcinoma genesis in a combined way and intervening with components extracted from Qi-boosting toxin-eliminating herbal medicines, nasopharyngeal mucosa was taken from the experimental animals and tissues samples were cut to do light and electronic microscopies respectindy in order to evaluated the pathohistological differences among these groups of animals. The results indicated that significantly increased number of animals with precancerous nasopharyngeal lesion was seen in the modeling group induced by exhausted swimming plus DNP using and markedly fewer number of animals with this kind of lesion was confirmed in the group simply using DNP to induce such a lesion (p<0.05).Fourthly, proteomic study was performed to screen and identify the associated proteins with the imitation and development of nasopharyngeal precancerous lesions among the animals affected of Qi-dificient status and DNP induced carcinogenesis among mile SD rats and determination of ultrastructural pathology on nasopharyngeal precancerous lesions among them, nasopharyngeal tissue samples were respectively taken from healthy control animals and ones with most significant precancerous lesion in the nasopharynx to extract the total cellular protein for electrophoresis in two dimensions(2-DE). Then, the gels were stained by Comas blue and analyzed by PDQuest software, with a match of normal nasopharyngeal tissue with that of precancerous lesions made on the dots of 2-DE gels. After this step, bio-informatics analysis was carried out to screen and identify the differentially expressed in the nasopharyngeal mucosa with precancerous lesion, 9 up-regulated protein were screened out, with a possible association in carcinogenesis, mainly invoking such molecular events as cell cycle regulation, protein transportation, energy metabolism, cellular adhesion and junction etc..In summary, Qi-deficient physique status is one physique status sensitive to environmental cancer-inducing materials such as DNP. This elevated sensitivity to cancer inducing effect by DNP suggested a molecular basis responsible for the precancerous lesion in the nasopharynx, possibly associated closely with the abnormal expression of Gsta genes and up regulated expression of IL-1βrelated genes. The interactive effects of Qi-deficient physique status with DNP using may iniciate the progressing process of nasopharyngeal precancerous lesions, causing very complex events in the signaling network take place underlying the carcinogenesis and involving lots of molecular events in this process.