本文選題：血瘀證 + TGF-β1��； 參考：《福建中醫(yī)藥大學(xué)》2014年博士論文

【摘要】：目的：基于TGF-β1信號(hào)通路探究感染乙肝病毒以后血瘀證患者病情由CHB發(fā)展至HBC乃至HCC的疾病進(jìn)展機(jī)制。 方法：本課題在文獻(xiàn)研習(xí)以及既往研究的基礎(chǔ)上,選擇TGF-β1信號(hào)通路作為目標(biāo)通路,通過(guò)對(duì)2例(血瘀證和脾虛證各1例)HBC發(fā)展至HCC患者的同一患者二個(gè)疾病階段的血清進(jìn)行蛋白細(xì)胞因子芯片以及各20對(duì)CHB、HBC和HCC血瘀證和脾虛證患者血清進(jìn)行代謝組學(xué)篩選復(fù)核比對(duì),再進(jìn)行定點(diǎn)蛋白和候選Micro檢測(cè)。 結(jié)果：1.蛋白芯片的細(xì)胞因子篩選發(fā)現(xiàn)了TGF-β1在血瘀證HCC患者較其HBC階段呈92.56的上調(diào)表達(dá),列細(xì)胞因子表達(dá)的第一位,遠(yuǎn)遠(yuǎn)高于脾虛證患者的20.1,提示TGF-β1可能不僅在疾病進(jìn)展中扮演重要角色,同時(shí)與血瘀證密切相關(guān),證實(shí)目標(biāo)通路的TGF-β1可以作為定點(diǎn)目標(biāo)蛋白,且能調(diào)控INFy的表達(dá)。2.代謝組學(xué)研究篩選出血瘀證的差異性物質(zhì)也與TGF-β1的異常表達(dá)相匹配,進(jìn)一步對(duì)目標(biāo)通路進(jìn)行復(fù)核證實(shí)；3.蛋白細(xì)胞因子表達(dá)驗(yàn)證提示血清中TGF-β1的蛋白表達(dá)以及TGF-β1/INFy值,HCCHBCCHBNormal (p0.01)。 INFy的蛋白表達(dá)則下調(diào)HCCHBCCHBNormal (p0.01)。以上結(jié)果由于數(shù)據(jù)方差不齊,均采用秩和檢驗(yàn)。TGF-β1/INFy結(jié)果趨勢(shì)同TGF-β1,由于通過(guò)方差齊性檢驗(yàn)采用LSD-t檢驗(yàn)表現(xiàn)出更好的趨向性上調(diào)表達(dá)(p0.01)。至于血瘀證與脾虛證患者的TGF-β1、INFγ、TGF-β1/INFy都匹配疾病進(jìn)展(p0.01),其中總血瘀證與總脾虛證比較,以及每個(gè)疾病間的血瘀證與脾虛證之間的比較都有顯著性差(p0.01)。4.Micro檢測(cè)：選擇的Micro也是與TGF-β1信號(hào)通路密切相關(guān),其中hsa-miR-21-5p在疾病三個(gè)階段患者較正常組都具有顯著差異(P0.01)；CHB患者表達(dá)上調(diào)最明顯,與HBC和HCC患者相比,也有顯著差異(P0.01)。hsa-miR-146a-5pCHB和HBC階段患者與正常人相比,有差異(P0.05), CHB患者表達(dá)上調(diào)與HCC患者相比也有差異(P0.05),同時(shí)HBC與HCC患者相比,差異顯著(P0.01)。 hsa-miR-29b-3pHCC與正常組有顯著差異(P0.01),表達(dá)明顯下調(diào)且與HBC有顯著差異(P0.01)和HBC亦有差異(P0.05)。至于血瘀證與脾虛證CHB與HCC階段無(wú)差異,在HCC階段hsa-miR-21-5p血瘀證和脾虛證具有顯著差異(P0.01)；hsa-miR-146a-5p血瘀證和脾虛證患者之間也有顯著差異(P0.01)。 結(jié)論：基于蛋白芯片、代謝組學(xué)復(fù)核以及定點(diǎn)蛋白和Micro技術(shù)驗(yàn)證我們發(fā)現(xiàn)乙肝病毒感染后血瘀證患者病情進(jìn)展與TGF-β1信號(hào)通路中蛋白和基因表達(dá)異常有關(guān)。而hsa-miR-29b-3p、hsa-miR-21-5p可能可以作為血瘀證的判別指標(biāo)。
[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 緯 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.
【學(xué)位授予單位】：福建中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R512.62

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