基于TGF-β1信号通路探究乙肝病毒感染后血瘀证患者病情进展的机制

发布时间：2018-05-27 18:14

本文选题：血瘀证 + TGF-β1　；参考：《福建中医药大学》2014年博士论文

【摘要】：目的：基于TGF-β1信号通路探究感染乙肝病毒以后血瘀证患者病情由CHB发展至HBC乃至HCC的疾病进展机制。方法：本课题在文献研习以及既往研究的基础上,选择TGF-β1信号通路作为目标通路,通过对2例(血瘀证和脾虚证各1例)HBC发展至HCC患者的同一患者二个疾病阶段的血清进行蛋白细胞因子芯片以及各20对CHB、HBC和HCC血瘀证和脾虚证患者血清进行代谢组学筛选复核比对,再进行定点蛋白和候选Micro检测。结果：1.蛋白芯片的细胞因子筛选发现了TGF-β1在血瘀证HCC患者较其HBC阶段呈92.56的上调表达,列细胞因子表达的第一位,远远高于脾虚证患者的20.1,提示TGF-β1可能不仅在疾病进展中扮演重要角色,同时与血瘀证密切相关,证实目标通路的TGF-β1可以作为定点目标蛋白,且能调控INFy的表达。2.代谢组学研究筛选出血瘀证的差异性物质也与TGF-β1的异常表达相匹配,进一步对目标通路进行复核证实；3.蛋白细胞因子表达验证提示血清中TGF-β1的蛋白表达以及TGF-β1/INFy值,HCCHBCCHBNormal (p0.01)。 INFy的蛋白表达则下调HCCHBCCHBNormal (p0.01)。以上结果由于数据方差不齐,均采用秩和检验。TGF-β1/INFy结果趋势同TGF-β1,由于通过方差齐性检验采用LSD-t检验表现出更好的趋向性上调表达(p0.01)。至于血瘀证与脾虚证患者的TGF-β1、INFγ、TGF-β1/INFy都匹配疾病进展(p0.01),其中总血瘀证与总脾虚证比较,以及每个疾病间的血瘀证与脾虚证之间的比较都有显著性差(p0.01)。4.Micro检测：选择的Micro也是与TGF-β1信号通路密切相关,其中hsa-miR-21-5p在疾病三个阶段患者较正常组都具有显著差异(P0.01)；CHB患者表达上调最明显,与HBC和HCC患者相比,也有显著差异(P0.01)。hsa-miR-146a-5pCHB和HBC阶段患者与正常人相比,有差异(P0.05), CHB患者表达上调与HCC患者相比也有差异(P0.05),同时HBC与HCC患者相比,差异显著(P0.01)。 hsa-miR-29b-3pHCC与正常组有显著差异(P0.01),表达明显下调且与HBC有显著差异(P0.01)和HBC亦有差异(P0.05)。至于血瘀证与脾虚证CHB与HCC阶段无差异,在HCC阶段hsa-miR-21-5p血瘀证和脾虚证具有显著差异(P0.01)；hsa-miR-146a-5p血瘀证和脾虚证患者之间也有显著差异(P0.01)。结论：基于蛋白芯片、代谢组学复核以及定点蛋白和Micro技术验证我们发现乙肝病毒感染后血瘀证患者病情进展与TGF-β1信号通路中蛋白和基因表达异常有关。而hsa-miR-29b-3p、hsa-miR-21-5p可能可以作为血瘀证的判别指标。
[Abstract]:Objective: to explore the mechanism of progression from CHB to HBC and HCC in patients with blood stasis syndrome after hepatitis B virus infection based on TGF- 尾 1 signaling pathway. Methods: TGF- 尾 1 signaling pathway was selected as the target pathway based on literature study and previous studies. The serum of 2 cases (1 case of blood stasis syndrome and 1 case of spleen deficiency syndrome) developed to the same disease stage of HCC patients were treated with protein-cytokine microarray and 20 pairs of serum metabolism of CHB HBC and HCC blood stasis syndrome and spleen deficiency syndrome respectively. Group screening check comparison, The target protein and candidate Micro were detected. The result is 1: 1. The cytokine screening of protein chip showed that TGF- 尾 1 up-regulated the expression of TGF- 尾 1 in HCC patients with blood stasis syndrome compared with their HBC stage, and the expression of TGF- 尾 1 was the first in the list of cytokines expression. This suggests that TGF- 尾 1 may not only play an important role in the progression of the disease, but also be closely related to blood stasis syndrome. TGF- 尾 1 in the target pathway can be regarded as a targeted target protein and can regulate the expression of INFy. Metabolic studies showed that the differential substances of blood stasis syndrome matched with the abnormal expression of TGF- 尾 1, and the target pathway was confirmed by further review. The expression of TGF- 尾 1 in serum and the 1/INFy value of TGF- 尾 in serum were confirmed by the expression of protein cytokines. The protein expression of INFy was down-regulated by HCCHBCCHBNormal p0.01. Because of the variance of the data, the results of rank sum test. TGF- 尾 1/INFy showed the same trend as that of TGF- 尾 1, and the LSD-t test showed a better tendency to up-regulate the expression of P0.01a through the test of homogeneity of variance. As for TGF- 尾 1 inf 纬 TGF- 尾 1/INFy in patients with blood stasis syndrome and spleen deficiency syndrome, TGF- 尾 1/INFy matched the progression of the disease, among which the total blood stasis syndrome was compared with the total spleen deficiency syndrome. There was significant difference between blood stasis syndrome and spleen deficiency syndrome in each disease. 4. Micro detection: the selected Micro was also closely related to TGF- 尾 1 signaling pathway. There was a significant difference in the expression of hsa-miR-21-5p between the three stages of the disease and the normal controls. There was also a significant difference in the upregulation of the expression of hsa-miR-21-5p between the patients with HBC and the patients with HCC, and the patients at the stage of P0.01hsa-miR-146a-5pCHB and the patients with HBC compared with the normal subjects. There were significant differences in the expression of P0.05 and P0.05in patients with CHB compared with those in patients with HCC, and in patients with HBC and HCC, there was a significant difference in the expression of P0.01and the expression of P0.01C in hsa-miR-29b-3pHCC and normal controls. The expression of hsa-miR-29b-3pHCC was down-regulated and significantly lower than that of HBC (P0.01) and HBC was also different (P0.05A). There was no difference between blood stasis syndrome and spleen deficiency syndrome CHB and HCC stage. In HCC stage, there was significant difference between hsa-miR-21-5p blood stasis syndrome and spleen deficiency syndrome. There was also a significant difference between patients with hsa-miR-146a-5p blood stasis syndrome and spleen deficiency syndrome. Conclusion: based on protein chip, metabonomics review and site-specific protein and Micro techniques, we found that the progression of blood stasis syndrome after hepatitis B virus infection is related to the abnormal expression of protein and gene in TGF- 尾 1 signaling pathway. Hsa-miR-29b-29b-3psa-miR-21-5p may be used as a discriminant index of blood stasis syndrome.
【学位授予单位】：福建中医药大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R512.62

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