Study On The Relationship Between The Gene Expression Of Energy Metabolism And The Syndrome Characteristics Of The Deficiency-cold Syndrome Or Deficiency-heat Syndrome

Objective：In this study, typical deficiency-cold syndrome patients anddeficiency-heat syndrome patients were chosen as the objects, The objective geneswere the differential genes which were came from early screening by gene expressionprofiles of candidate. Those differential genes of candidate expression were detectedby the real-time fluorescence quantitative PCR to explore the difference of energymetabolism between deficiency-cold syndrome and deficiency-hot syndrome, then therelation between characteristic of syndrome and unusual expression of genes wererevealed, to probe the influence and law which through change the environmenttemperature on productivity function of different syndrome organism. The relativitywas expected to find between the unusual energy metabolism, the occurring ofdeficiency-cold syndrome and deficiency-heat syndrome in the molecular level. Andthis will lay the foundation for revealing the scientific connotation of deficiency-coldsyndrome and deficiency-heat syndrome in micro-level.Methods: With Chinese constitution Scale to survey the students who werepresent in Henan University of TCM, then analysis the distribution of constitution ofthem. people who had the typical syndrome were diagnosed through expert, anddivided into deficiency-cold syndrome group, deficiency-heat syndrome group andnormal control group of20. Real-time fluorescence quantitative PCR was used toquantity the P2X5receptor, SLC4A8, complement C5, FABP6, PEPP1and IDH1,IDH2, IDH3A, IDH3B, IDH3G in different temperature, namely one group at room,and the other at cold temperature (low temperature-4℃～5℃, room temperature23℃～25℃). Application of SPSS17.0statistical software for statistical processing,compared among the three groups using the analysis of variance,Paired samples T-testwas used to compare Groups between the two temperatures. Results1.Epidemiological survey results of TCM constitution: the distribution of eightbiased constitutions(sum of simple and combine): Qi deficiency29.3%; yangdeficiency25.2%; yin deficiency,24%;dampness-phlegm13.4%;damp-heat9.9%;blood stasis14.1%; qi stagnation18.6%. Further analysis of deficiency of yin and yangphysique:758cases with constitution of Yang-deficiency were found(25.2%)，in whichthere were332girls and423boys，the difference of the composition ratio in differentgender wasn’t significant(P＞0.05), and feeling cold with hands and feet，wanted moreclothes and couldn’t bear cold were the main manifestations. There were310girls and412boys in722cases of Yin-deficiency constitution(24%), and the difference of thecomposition ratio in different gender was significant(P＜0.05).The principalmanifestations were lip and eyes feeing dry and thirst．2.The expression of P2X5receptor in deficiency-cold syndrome anddeficiency-heat syndrome groups was lower than that in the normal control group atroom temperature(P＜0.05); the difference of P2X5receptor expression betweennormal group and deficiency-cold syndrome group, as well as that betweendeficiency-cold syndrome group and deficiency-heat syndrome group was statisticallysignificant at low temperature. In group comparison, the expression of P2X5receptorhave no differences in all groups at the two different temperatures (P>0.05).3.The expression of SLC4A8gene:○1Under either room temperature or lowtemperature environment, SLC4A8gene expression of deficiency-cold syndrome werehigher than the other two groups.The difference between normal group anddeficiency-cold syndrome group was statistically significant at lowtemperature(P=0.015<0.05).○2In group comparison:SLC4A8Gene expression ofdeficiency-cold syndrome group and deficiency-heat syndrome group under lowtemperature was higher than that room temperature. And SLC4A8Gene expression ofthe normal control group under low temperature lower than that room temperature.These differences were all no statistically significant.4.(1)IDH1gene expression:○1Under either room temperature or low temperatureenvironment, IDH1gene expression of deficiency-heat syndrome group was thehighest, and that was the lowest in the normal control group. The differences betweenthe normal control group and deficiency-cold/heat syndrome group were statisticallysignificant at room temperature(P=0.031＜0.05，P=0.003＜0.05).The differencesbetween the normal control group and deficiency-heat syndrome group were statistically significant at low temperature.○2In group comparison: IDH1geneexpression was higher under room temperature than that under low temperature in agroup. And the difference was statistically significant in deficiency-heat syndromegroup(P=0.008＜0.05).(2)IDH2gene expression:○1Under either room temperature orlow temperature environment, IDH2gene expression in the normal control group washighest, and that was lowest in deficiency-heat syndrome group. These differenceswere all no statistically significant(P＞0.05).○2In group comparison: It is under lowtemperature that IDH2gene expression in three groups was higher. However thedifferences were all no statistically significant(P＞0.05).(3)IDH3A geneexpression: IDH3A gene expression in deficiency-cold syndrome group was lowerthan that in other two group under either room temperature or low temperatureenvironment. And the differences were all statistically significant under lowtemperature environment. In group comparison: Compared to room temperature,IDH3A gene expression in deficiency-cold syndrome group and the normal controlgroup were lower under low temperatures.The difference was statistically significant indeficiency-cold syndrome group(P=0.004<0.05).The differences in others were nostatistically significant(P＞0.05).(4)IDH3B gene expression: IDH3B gene expressionin deficiency-cold syndrome group was higher than that in others under roomtemperature. And these differences were statistically significant(P=0.03<0.05,P=0.037<0.05). The differences in other two groups were no statisticallysignificant(P>0.05). In group comparison:It is under room temperature that IDH3Bgene expression was higher than that under low temperature in deficiency-coldsyndrome group and in the normal control group. And the difference in deficiency-coldsyndrome group was statistically significant(P=0.008<0.05). IDH3B gene expressionwas higher under low temperatures in deficiency-heat syndrome group. However thedifferences was no statistically significant(P>0.05).(5)IDH3G gene expression:Compared to the normal control group, the differences of IDH3G gene expression indeficiency-cold syndrome group and deficiency-heat syndrome group were nostatistically significant(P>0.05). And that in deficiency-cold syndrome group washigher than other two groups. The difference between deficiency-cold syndrome groupand deficiency-heat syndrome group was statistically significant(P=0.013<0.05).IDH3G gene expression was highest in deficiency-heat syndrome group, lowest indeficiency-heat syndrome group under low temperatures.And the differences were nostatistically significant(P>0.05). In group comparison: It is under low temperature that IDH3G gene expression was lower than that under room temperature indeficiency-cold syndrome group and in the normal control group.And the differenceswere statistically significant(P=0.022＜0.05, P=0.037＜0.05). However it wasopposite in deficiency-heat syndrome group, and the difference was no statisticallysignificant(P>0.05).5. The expression of Complement Component5:○1At room temperature, the C5gene expression in three groups as follows: Deficiency-cold syndrome＞normalcontrol group＞Deficiency-heat Syndrome, the differences were statisticallysignificant(P=0.001＜0.05，P=0.000＜0.05，P=0.005＜0.05)；At low temperature,thegene expression in three groups as follows:Deficiency-heat Syndrome＞normal controlgroup＞Deficiency-cold syndrome,differences had no statistical significance(P＞0.05);○2In group comparison: Expression at low temperatures were lower than roomtemperature in normal control group and deficiency-cold syndrome,the comparison ofdeficiency-cold syndrome at low temperature and room temperature difference hadstatistical significance(P=0.000＜0.05); but in deficiency-heat Syndrome,theexpression at low temperature was higher than room temperature(P＞0.05).6. The expression of FABP6:○1The expression of FABP6in deficiency-coldsyndrome was higher than deficiency-heat Syndrome and normal control groups atroom temperature,deficiency-heat Syndrome was lowest expression. Deficiency-heatSyndrome compared with normal group,compared with the deficiency-coldsyndrome,the differences were statistically significant(P=0.031＜0.05， P=0.025＜0.05).Deficiency-cold syndrome and the normal group, the difference was notstatistically significant (P＞0.05);At low temperature,the gene expression in threegroups as follows:Deficiency-heat Syndrome＞Deficiency-cold syndrome＞normalcontrol group,differences had no statistical significance(P＞0.05).○2In groupcomparison: Expression at low temperatures were lower than room temperature innormal control group and deficiency-cold syndrome, the differences were statisticallysignificant(P=0.017＜0.05， P=0.01＜0.05);but in deficiency-heat Syndrome,theexpression at low temperature was higher than room temperature(P＞0.05).7. The expression of PEPP1:○1At room temperature, PEPP1gene expression inthree groups as follows: normal control group> Deficiency-cold syndrome＞Deficiency-heat Syndrome; at cold temperature,the gene expression in three groups asfollows: Deficiency-cold syndrome＞normal control group＞Deficiency-heatSyndrome,differences had no statistical significance(P＞0.05);○2In group comparison:Three groups of people in two temperature were characterized by lowtemperature＞room temperature,but the differences had no statistical significance(P＞0.05).Conclusion:1.It is necessary to pay more attention about the physical problems of collegestudents according to the distribution of eight pathological constitutions.2.The typical syndrome characteristics of Deficiency-cold syndrome andDeficiency-heat Syndrome may be associated with the expression of P2X5receptorand SLC4A8.3.IDH genes were different expression in Deficiency-cold syndrome andDeficiency-heat Syndrome. Key genes associated with energy generation wereabnormal expression.Meanwhile, The H+is also flow out. These might be related to thecharacteristics of the syndrome.4.Complement C5in Deficiency-cold syndrome patients was abnormal expressionin low temperature lead the immune defence were not properly understood,so sickmore easily at low temperature.5.It is not enough evidence that the FABP6and PEPP1abnormal expressionimpact on the characteries of Deficiency-cold syndrome and Deficiency-heat syndrome6.Since the function of different genes,the sensitive to temperature were alsodifferent,these intrinsic mechanism to be studied.The effect of temperature changes onenergy metabolism remains to be further defined.