| Object:In this study, our aim was to investigate the expression of CXCL6and its receptors (CXCR1and CXCR2) in peripheral blood and liver biopsies of chronic hepatitis B (CHB) and cirrhosis patients, analysis the relationship between CXCL6and HBV-DNA contains, AST and ALT level. Explore the role of CXCL6, CXCR1and CXCR2in HBV-induced chronic hepatitis and cirrhosis in the molecular mechanisms.Methods:The serum CXCL6level of37patients with typical CHB,17patients with cirrhosis and21control (normal blood donors) was detected by ELISA; RT-PCR was used to investigate the mRNA expression of CXCL6, CXCRl and CXCR2, individual values of mRNA were normalized to GAPDH mRNA and the final mRNA level was represent as lgcDNA/1gGAPDH. SP immunohistochemistry stain was used to observe the expression of CXCL6, CXCR1and CXCR2in liver biopsy from9cases of CHB,14cases of cirrhosis (9compensated and5decompensated) and12cases of patients with Hepatic carcinoma.17cases of normal liver tissue from liver trauma surgery were used as normal control.Results:Expression of GCP-2in serum of patients with CHB and cirrhosis were (673.11±199.57)pg/ml and (736.59±181.87)pg/ml, respectively, significantly higher than normal control (P<0.01). The serum level of GCP-2of HBV-DNA (+) group and HBV-DNA (-) group were (810.40±170.45)pg/ml and (546.46±101.83) pg/ml, the difference between the two groups was statistically significant (P<0.01). The expression of CXCL6in peripheral blood of patients with CHB and cirrhosis were (0.616±0.279)1gcDNA/1gGAPDH and (0.871±0.209)1gcDNA/1gGAPDH, respectively, significantly higher than normal control (P<0.01). The expression of CXCL6was related to the HBV loading. HBV-DNA (+) group ((0.774±0.276)1gcDNA4gGAPDH) was significantly higher than HBV-DNA (-) group (P=0.0329). The expression of CXCL6in patients with cirrhosis was significantly higher than which in patients with CHB. The CXCR1mRNA level in peripheral blood of patients with CHB and cirrhosis were (0.770±0.219)lgcDNA/1gGAPDH and (0.808±0.299)lgcDNA4gGAPDH respectively, and have significantly differences with the control group (P<0.01). There’s no significantly difference between group CHB, cirrhosis, HBV-DNA (+) and HBV-DNA (-)(P>0.05). The difference of the expression of CXCR2mRNA between the patients with CHB, cirrhosis and normal control was not statistically significant (P>0.05). The ALT and AST level in serum of patients with CHB were (63.17±21.46) U/L and (54.78±26.15) U/L, respectively. The ALT and AST level in serurn of patients with cirrhosis were (69.44±24.51) U/L and (72.19±22.64) U/L, respectively. The correlation analysis showed that the HBV-DNA loading in patients with CHB and cirrhosis was significantly correlated with serum CXCL6levels (r=0.774), peripheral blood CXCL6mRNA content (r=0.558) and peripheral blood CXCR1mRNA content (r=0.627) when P<0.01, and was significantly correlated with CXCR2mRNA content (r=0.315) when P<0.05. The level of ALT and AST in serum of patients with CHB and cirrhosis were significantly correlated with serum CXCL6levels (r=0.757, r=0.745), peripheral blood CXCL6mRNA content (r=0.577, r=0.565) and peripheral blood CXCRlmRNA content (r=0.633, r=0.637) when P<0.01. There’s no significant correlation between ALT nor AST and CXCR2mRNA content (r=0.256, r=0.250) even when P<0.05. The SP immunohistochemical showed that the positive rate of the expression of CXCL6in patients with CHB, cirrhosis and Hepatic carcinoma were76.47%,78.57%and83.33%, respectively. The positive rate of the expression of CXCL6in patients with CHB, cirrhosis and Hepatic carcinoma were significantly higher than that of the control group, the difference was statistically significant (P<0.01). The positive rate of the expression of CXCR1in patients with CHB, cirrhosis and Hepatic carcinoma were82.35%,78.57%and91.67%, respectively, which were obviously higher than the control(P<0.01). The positive rate of the expression of CXCR2in patients with CHB, cirrhosis and Hepatic carcinoma were52.94%,50%and58.33%, respectively, there’s no statistically difference between the positive rate of the expression of CXCR2in patients with CHB, cirrhosis, Hepatic carcinoma and control(P>0.05). Liver tissue RT-PCR showed that the CXCL6mRNA level of patients with CHB, cirrhosis and Hepatic carcinoma were (0.675±0.282)1gcDNA/1gGAPDH,(0.883±0.230)1gcDNA4gGAPDH and (1.055±0.175)1gcDNA4gGAPDH, respectively, as compared with the normal control, the difference were statistically significant (P<0.01). The CXCL6mRNA level of liver tissue from Hepatic carcinoma was significantly higher than that from cirrhosis and CHB (P<0.05). The CXCR1mRNA expression level of liver tissue from CHB, cirrhosis and Hepatic carcinoma were (0.685±0.160)1gcDNA/1gGAPDH, (0.854±0.232)1gcDNA/1gGAPDH and (1.050±0.183)1gcDNA4gGAPDH respectively, as compared with the normal control, the difference were statistically significant (P<0.01). The CXCR1mRNA expression level of liver tissue from Hepatic carcinoma was significantly higher than that of cirrhosis and CHB(P<0.05). The CXCR2mRNA expression level of liver tissue from CHB, cirrhosis and Hepatic carcinoma were (0.481±0.159)1gcDNA/1gGAPDH,(0.502±0.116)1gcDNA4gGAPDH and (0.490±0.144)1gcDNA4gGAPDH, the difference was not statistically significant (P>0.05).Conclusion:The serum GCP-2level of patients with chronic hepatitis B and cirrhosis was significantly increased while compared with normal blood donors. The expression of CXCL6mRNA and CXCR1mRNA in peripheral blood and liver tissue from patients with chronic hepatitis B and cirrhosis were higher than which from control. The expression of CXCL6mRNA and CXCR1mRNA was positively correlated with HBV-DNA loading and the level of ALT and AST. There’s no significant difference between chronic hepatitis, cirrhosis and normal control on the expression of CXCR2mRNA. All these result demonstrate that CXCL6and CXCR1may be associated with the development of chronic hepatitis and cirrhosis. SP immunohistochemical showed CXCL6expression was significantly higher in patients with Hepatic carcinoma, indicate that CXCL6likely contribute to the occurrence of liver cancer. |
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