Dampness Constitution Obesity Subtypes And Prone To Metabolic Syndrome, Oxidative Stress Mechanisms

Experiment 1 Compare the diagnoses of Phlegmy-wet constitution with the traditional method of HOMA-IR in screening of MHO and MONW person[Background] With the rapid development of global economy and the improvement of people's standard of living, obesity in cities and towns has become a concern of the public health field at home and abroad. The development to the depth give impetus to the constant revision of obesity's diagnosis standard from weight, body lipid rate, to body mass index(BMI), waistline. From the diagnosis standard, we can see the perplexity in research, which is that obesity is only superficial phenomenon, what is the representation of essence? The diagnoses above are all stay on the level of morphology, which can't descript the potential connotation. That is the reason of "obesity subtypes", obesity subtypes include metabolically healthy but obese (MHO) and metabolically obese, narmal-weight (MONW). Phlegmy-wet constitution is a biased constitution whith the nature of heavy, sticky and stagnant. Based on the fore-working Phlegmy-wet constitution and obesity are closely related, which may be the background and foreground factors of Metabolic Diseases. Although the morphology description of Phlegmy-wet constitution is obesity, it is not equal to obesity. Phlegmy-wet constitution is not a simple concept of morphology, but implies the pathophysiological mechanism of body fluid conveyance obstacles and phlegm-damp retention. Therefore we propose that the diagnosis of phlegmy-wet constitution is more suitable for the identification of metabolic disorder population. Combined with the concept of obesity subtypes, Phlegmy-wet constitution may be the internal factors of subtypes phenomenon of obesity.[Objective] Study on the feasibility of screening of MHO and MONW persons using the diagnose of Phlegmy-wet constitution. Reveal the mechanismin of obesity subtypes in the view of constitution, so as to a more profound understanding of obesity.[Method] 1. The Four table match chi-square test between the two diagnoses (HOMA-IR and Phlegmy-wet constitution) in obesity people; 2. T test between the Phlegmy-wet non-obese people and normal constitution people; 3. Logistic regression analysis of hazards related to IR; 4. Logistic regression analysis of hazards related to Phlegmy-wet constitution. [Results] 1. There was no significant difference in relevance ratio among the two diagnoses; 2. The levels of IR was higher in the Phlegmy-wet non-obese people than normal constitution person; 3. Multiple logistic regression analysis revealed that Phlegmy-wet constitution, waistline, family inheritance history and drink were dangerous factors for Phlegmy-wet constitution; 4. Multiple logistic regression analysis revealed that family inheritance history and greasy dietary habit were dangerous factors for Phlegmy-wet constitution.[Conclusion] It is feasible to screen of MHO and MONW persons using the diagnose of Phlegm-wet constitution, even more sensitive than the traditional method of HOMA-IR. Multiple logistic regression analysis further revealed that Phlegm-wet constitution is a independent factor influencing degree of IR. And it is influenced by Natural Endowment and Improper living habits rather than weight. Phlegm-wet constitution is the internal factors of obesity subtypes,whith was more suitable for the identification of metabolic disorder population. Experiment 2 the oxidative stress mechanism contributing to the Susceptibility of Phlegm-wet constitution to Metabolic Syndrome[Background] Phlegm-wet constitution and metabolic disorders are closely related. Phlegm-wet constitution persons are the high risk group of Metabolic Syndrome (MS) based on the fore-work of clinical epidemiology investigation. Based on the results of experiment 1, we propose that the physiological pathology mechanism of body fluid conveyance obstacles and phlegm-damp retention is common soil of Various components of MS. In recent years the oxidative stress mechanism is paid more attention in IR and metabolic syndrome. Oxidative stress is related to many kinds of factors such as outside environment, the phenomenon of diet and living environment, and present earlier before clinical symptoms, which is the connection of risk factors and IR. It can provide a powerful molecular basis for early detection and intervention therefore, we propose that Phlegm-wet constitution may result in IR and MS through oxidative stress mechanism.[Objective] Study on the mechanism of Phlegm-wet constitution related to MS. Enrich the micromechanism of Phlegm-wet constitution related to diseases. provide evidence for Prevention of the metabolic syndrome in the view of constitution.[Method]1. Object of study are normal constitution group(PI)20 people, the Phlegm-wet non-obese group(TF)20, Obesity non-phlegm-wet group(FF)25 people and Obesity phlegm-wet group(FT)30people, and MS group25people,using the Baseline of Age, gender and BMI. Compare the general data in different groups; 2. Compare the serum FFA levels and IR in different groups; 3. Compare the serum T-AOC and activity of SOD,GSH-Px,CAT in different groups and the levels of mRNA in PBMC; 4. Observe change of GSH/GSSG in different groups; 5. Compare the serum NO levels and activity of NOS,iNOS in different groups;6. Compare the serum oxidative damage product levels such as MDA,ox-LDL, and 8-OHdG; 7. correlation test of indexes above and Phlegm-wet constitution score and IR.[Result] 1. The baselines of the general data and general biochemical indicator were no different among the 5 groups, except MS group had a higher levels of biochemical indicator; 2. The levels of FFA and IR were significantly higher in Phlegm-wet constitution group no matter it is whether obesity or not (P< 0.05); 3. The levels of T-AOC, CAT were significantly higher in Phlegm-wet constitution group no matter it is whether obesity or not. SOD was decreased oppositely; 4. GHS/GSSG is inclined to oxidized direction excursi in Phlegm-wet constitution group and MS group; 5. The levels of NO were decreased in Phlegm-wet constitution group no matter it is whether obesity or not, but this was not statistically significant (P> 0.05); The levels of NO were increased in MS group; 6. The levels of MDA,ox-LDL, and 8-OHdG were increased in Phlegm-wet constitution group and MS group, with a statistical significancepartly; 7. A positive correlation between score of Phlegm-wet constitution and IR in no-patient group, but there is no significant correlation in MS group. IN no ill stage, T-AOC,CAT,FFA,iNOS were positive correlation with score of Phlegm-wet constitution and IR, SOD,GSH-Px,GSH/GSSG,NO,MDA,OX-LDL 8-OHDG were negative correlation with score of Phlegm-wet constitution and IR. In MS stage, T-AOC,CAT were negative correlation with IR oppositely, NO was positive correlation with IR oppositely.[Conclusion] 1. Obesity could promote the extent of Phlegm-wet and oxidative stress, but there was no causal relationship between them; 2. Early sensitive index of Phlegm-wet constitution is SOD, which may be related with natural endowment; 3. Phlegm-wet constitution people had oxidative damage even if they are not ill; 4. The oxidative stress was not only quantity accumulation in constitution and desaeses, but also present transition in oxidation resistance. Namely for biased constitution and disease stage were not only have difference in the amount, but also have essential difference, which may provided a reference for the study of constitution and syndrome in TCM. Experiment 3 Analysis on differential expression gene and signal pathway related to oxidative stress in phlegm-wet constitution by gene chip technology[Objective] Give an exploratory analysis on differential expression gene and related signal pathway which related to oxidative stress, to provide basi c information for carrying out animal experiment and medicine intervention on the molecular mechanism of Phlegm-wet constitution related to disease.[Method] Object of study are normal constitution group16 people and th e Phlegm-wet constitution group 16 people. The total RNA was extracted fr om isolated peripheral blood monocytes, and the whole-profile of gene expr ession was detected by Affymetrix genechip human genome U133 plus 2.0. Analyse gene and differences signal pathway related to oxidative stress, sta tistical standard is P< 0.05 or (and) Fold change≥2 or≤0.5.[Result] 1. SOD2 gene Corresponding to SOD was down-regulated in phlegm-wet constitution group, which close to significant (P=0.058), and is consistent with the protein level. GPX3, CAT gene corresponding to GSH-PX, CAT were down-regulated, but having no significant.2. NF-κB signaling pathway:NF-κB1 gene was significantly up-regulated(Fold change=1.16; P< 0.01). PKC signaling pathway:PRKCβwas significantly up-regulated(Fold change=1.23; P<0.01)[Conclusion] 1. Impaired antioxidant was associated with phlegm-wet cons titution itself based on experiment 2. SOD was the sensitive index. The re sult of experiment 3 showed that SOD corresponding gene was down-regulat ed, which furtherly support this view above. Therefore, SOD gene may be o ne of the Congenital genetic sensitive indexs of phlegm-wet constitution. Se nsitivity of other antioxidative enzymes was not strong.2. Stress factor ox-LDL,NO may activate NF-κB signaling pathway in phlegm-wet constitution. NF-κB signaling pathway is closely related with IR and damaged Insulin s ecretion, which is the classic pathway of MS. The result furtherly support that phlegm-wet constitution is susceptible to MS.3. Stress factor may acti vate PKC-PKD signaling pathway. PKC-PKD signaling pathway is also close ly related with MS. It is the important is important molecular pathway parti cipating in Endothelial dysfunction of diabetes mellitus and hypertension. It is also important targets of medicine intervention, which can provide basic information for carrying out phlegm-wet constitution intervention study.