| BackgroundOur research team has been using the gene chip technology to test the microRNA expression profiles in AIDS patients with Spleen-Kidney Yang Deficiency Syndrome, Heat-toxic Stagnation Syndrome and healthy volunteers. Spleen-Kidney Yang Deficiency Syndrome group compared with the healthy control group, we screened out 89 microRNAs of which the expression had significant difference(P<0.05), 52 up-regulated(58%),37 down-regulated(42%). GO enrichment analysis showed that the biological basis of the syndrome may be associated with natural killer cell mediated cytotoxicity, the role of the IL-15, IL-6 receptor during T cell activation, apoptosis, transforming growth factor beta(TGF-β) and other related mechanism. Heat-toxic Stagnation Syndrome group compared with the healthy control group, we screened out 100 microRNAs of which the expression had significant difference(.P<0.05),64 up-regulated(64%),36 down-regulated(36%), GO enrichment analysis showed that the biological basis of the syndrome may be related to the role of IL-21 receptor during the activation of T cells, platelet, C reactive protein, oxidative stress and so on.ObjectiveTo investigate the correlation between the expression of different microRNA and AIDS in Spleen-Kidney Yang Deficiency Syndrome and Heat-toxic Stagnation Syndrome, and to find the microRNA specific expression characteristics in AIDS syndromes, to explore the biological basis of AIDS Spleen-Kidney Yang Deficiency Syndrome and Heat-toxic Stagnation Syndrome, and provide experimental evidence for the objective TCM syndrome diagnosis of AIDS.MethodsThis study carried out 9 microRNAs that significant differently expressed in AIDS Spleen-Kidney Yang Deficiency Syndrome and 9 microRNAs that significant differently expressed in Heat-toxin Stagnation Syndrome from the results of microarray study, including 5 of them were common to both syndromes(The target gene in Spleen-Kidney Yang Deficiency Syndrome:hsa-let-7b-5p, hsa-miR-15b-5p, hsa-miR-19b-3p, hsa-miR-30e-5p, hsa-miR-106b-5p, hsa-miR-144-3p, hsa-miR-144-5p, hsa-miR-320a, hsa-miR-320c; The target gene in Heat-toxin Stagnation Syndrome: hsa-let-7b-5p, hsa-let-7f-1-3p, hsa-let-7g-5p, hsa-miR-15b-5p, hsa-miR-126-3p, hsa-miR-144-3p, hsa-miR-320a, hsa-miR-320c, hsa-miR-3940-5p; The target gene repeated in both syndromes:hsa-let-7b-5p, hsa-miR-15b-5p, hsa-miR-144-3p, hsa-miR-320a and hsa-miR-320c). The expression of the target microRNAs in the samples from AIDS patients with Spleen-Kidney Yang Deficiency Syndrome, Heat-toxin Stagnation Syndrome and health volunteers were measured by Quantitative Real-time PCR, with U6 as the internal reference,2-△CT values as the relative expression of target gene. The expression of target microRNAs in AIDS Spleen-Kidney Yang Deficiency Syndrome group, Heat-toxic Stagnation Syndrome group and healthy control group were compared with SPSS V20.0 statistical software.Results1 The Spleen-Kidney Yang Deficiency Syndrome group compared with healthy control group:the expression of hsa-miR-19b-3p, hsa-miR-15b-5p, hsa-miR-30e-5p, hsa-miR-106b-5p, hsa-miR-144-5p are down-regulated in AIDS Spleen-Kidney Yang Deficiency Syndrome group, there are significant statistical difference(P<0.05), and are consistent with the microarray results.The expression of hsa-let-7b-5p, hsa-miR-144-3p, hsa-miR-320a are down-regulated in AIDS Spleen-Kidney Yang Deficiency Syndrome group, there are significant statistical difference(P<0.05). The expression of hsa-miR-320c is down-regulated in AIDS Spleen-Kidney Yang Deficiency Syndrome group, there aren’t significant statistical difference(P>0.05). The expression of the 4 microRNAs aren’t consistent with the microarray results.2 The Heat-toxin Stagnation Syndrome group compared with healthy control group: the expression of hsa-let-7g-5p, hsa-miR-126-3p are down-regulated in AIDS Heat-toxin Stagnation Syndrome group, there are significant statistical difference(P<0.05), and are consistent with the microarray results.The expression of miR-15b-5p is down-regulated in AIDS Heat-toxin Stagnation Syndrome group, and the downward tendency is consistent with the microarray result, but there isn’t significant statistical difference(P>0.05).The expression of hsa-let-7b-5p, hsa-miR-144-3p, hsa-miR-320a, hsa-miR-320c, hsa-miR-3940-5p are down-regulated in AIDS Heat-toxin Stagnation Syndrome group, there are significant statistical difference(P<0.05). The expression of hsa-let-7f-1-3p is down-regulated in AIDS Spleen-Kidney Yang Deficiency Syndrome group, there isn’t significant statistical difference(P>0.05). The expression of the 6 microRNAs aren’t consistent with the microarray results.3 The expression of hsa-let-7b-5p, hsa-miR-15b-5p, hsa-miR-144-3p, hsa-miR-320a, hsa-miR-320c have no difference between AIDS Spleen-Kidney Yang Deficiency Syndrome group and Heat-toxin Stagnation Syndrome group, there aren’t significant statistical difference(P＞0.05).Conclusions1 The results of hsa-miR-19b-3p, hsa-miR-30e-5p, hsa-miR-106b-5p, hsa-miR-144-5p in qRT-PCR are consistent with the microarray results. The results in the study show that these 4 microRNAs have certain correlation with AIDS Spleen-Kidney Yang Deficiency Syndrome, and suggest that they might be the biomarkers for AIDS Spleen-Kidney Yang Deficiency Syndrome, which possibly related to the regulation in immunoregulation and inflammatory reaction might be the biological basis of AIDS Spleen-Kidney Yang Deficiency Syndrome, such as IL-6, natural killer cell activity, TGF-p, tumor necrosis factor(TNF-a).2 The results of hsa-let-7g-5p and hsa-miR-126-3p in qRT-PCR are consistent with the microarray results. The results in the study show that these 2 microRNAs have certain correlation with AIDS Heat-toxin Stagnation Syndrome, and suggest that they might be the biomarkers for AIDS Heat-toxin Stagnation Syndrome, which possibly related to the regulation in biological processes might be the biological basis of AIDS Heat-toxin Stagnation Syndrome, such as cell apoptosis, endothelial dysfunction and immune response.3 The expression of hsa-let-7b-5p, hsa-miR-144-3p, hsa-miR-320a in the study are different between AIDS Spleen-Kidney Yang Deficiency Syndrome group and healthy control group, also in the comparison between AIDS Heat-toxin Stagnation Syndrome group and healthy control group, and have no difference between AIDS Spleen-Kidney Yang Deficiency Syndrome group and Heat-toxin Stagnation Syndrome group, which remind the 3 microRNAs have important implications for the AIDS, might be the potential biomarkers in AIDS. |
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