Mechanism Of TGF-β1Signal Transduction Pathway Of Hepatitis B Virus Infection Disease In Patients With Blood Stasis Syndrome

Objective:To explore the disease progression mechanism (infected by HBV virus, from CHB to HBC even to HCC) caused by blood stasis syndrome from the molecular biology perspective, verifying the assumption that blood stasis is the main factor leading to the disease progression.Methods:Based on previous researches, this paper adopted two sample cases whose diseases have developed to HCC from HBC (Choose TGF-β1signal transduction pathway as the target transduction pathway). One of the cases is a spleen deficiency syndrome patient, and the other one is a blood stasis syndrome case. Protein chip detections were carried out due to the serums of each patient. The serums are collected both in their HBC stage and HCC stage. Besides, metabonomics screenings are also implemented based on3groups of patients based on twenty pairs of spleen deficiency syndrome and blood stasis patients. They were divided into three groups which are CHB, HBC and HCC as well. Make the result comparison and revision between two experiments, figuring out the cytokines relating to blood stasis in previous research. Finally, fixed-point protein and candidate Micro detection are made.Results:1. Protein chip cytokines screening detected that, compared to HBC blood stasis syndrome patients, TGF-β1, ranking first in cytokine expression, shows92.56up-regulation expression in HCC blood stasis syndrome patients. This expression is much higher than that of HBC spleen deficiency patients (The expression for HBC spleen deficiency patients is20.1). This suggests that TGF-β1may not only play a leading role in disease progression but also have close relationship with blood stasis syndrome, show how blood stasis affect the disease progression and adjust the INFy expression.2. Different materials of blood stasis syndrome, screened by metabolomics, matched the abnormal expression of TGF-β13. Protein cytokines suggested that the protein expression of TGF-β1in serum and the value of TGF-β1/INFy is patterned as HCC>HBC>CHB>Normal(p<0.01). The protein expression of INFy is down regulation.(HCC<HBC<CHB<Normal (p<0.01)). As the variance is non-homogenous, the outcomes above are based on rank sum test. The trend of the result of TGF-β1/INFy is similar to that of TGF-β1. As the variance for TGF-β1/INFγ is homogenous, T test is carried out, which perfectly showed the tendency of increase expression(p<0.01). TGF-β1, INFy, TGF-β1/INFγ for both blood stasis syndrome and spleen deficiency syndrome patients all go with the disease progress (p<0.01).4. In Micro Test:hsa-miR-21-5p has a significant difference between patients of three different stages and the normal people. The CHB patient has the most apparent up-regulation expression, which also have significant variation compared to HBC and HCC patients (P<0.01). As for hsa-miR-146a-5p, apart from normal people, patients from CHB and HBC stage are variant (P<0.05). The up-regulation expressions of the CHB patients have the difference from HCC patients (P<0.05) as well. Furthermore, the difference of expression between HBC and HCC is significant (P<0.01). hsa-miR-29b-3p of HCC group is significantly different from the normal group (P<0.1). The hsa-miR-29b-3p expression of HCC is evidently down-regulation, which has significant variant with HBC (P<0.01) and different from HBC (P<0.05). There is not difference between blood stasis syndrome and spleen deficiency syndrome during CHB and HCC stages. In the HCC stage, hsa-miR-21-5p for blood stasis syndrome group and spleen deficiency syndrome group has significant variance (P<0.01). hsa-miR-146a-5p also have significant difference between blood stasis syndrome group and spleen deficiency syndrome group (P<0.01).Conclusion:Based on molecular biological techniques, the conclusion can be drawn that after the infection of HBV virus, blood stasis is the main factor that leads to the disease progression. Mechanism of blood stasis is related to the TGF-β1protein and Abnormal gene expression, and may be related to the TGF-βsignal path.Based on protein chip, metabonomics review and fixed-point protein and Micro technology validation we found that the progress of hepatitis B virus infection disease patients with blood stasis syndrome and TGF-β1protein and gene expression of signal pathway abnormity. Hsa-miR-29b-3p、hsa-miR-21-5p can be regard as the evaluation index of blood stasis syndrome.