摘要
目的:探讨hFgl2凝血酶原酶(hunman Fibrinogen-like protein 2,人Fgl2凝血酶原酶)在肝癌及病毒性肝炎进展机制中的作用。研究hFgl2凝血酶原酶阳性表达的细胞种类及其与肝功能总胆红素(TBil)、丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)之间的相关性以判断其对肝细胞坏死程度反映的敏感性,从而了解hFgl2凝血酶原酶的表达测定的临床意义。
方法:将观察对象分为A、B、C、D、E、F六组,取健康体检对照组20例(A组)及慢性乙型肝炎患者37例(包括B组:慢性乙型肝炎轻度15例,C组:慢性乙型肝炎中度13例,D组:慢性乙型肝炎重度9例),慢性重型乙型肝炎20例(E组),肝癌患者21例(F组)的外周血提取单核细胞行hFgl2凝血酶原酶蛋白(简称hFgl2蛋白)免疫组化制片;并对配对肝癌及慢性乙型肝炎患者肝组织进行hFgl2蛋白免疫组化制片,观察采集到的hFgl2蛋白表达阳性单核细胞及肝细胞图像中阳性表达部位及强度,并使用MIAS医学图像分析软件测定反映hFgl2蛋白表达情况的灰度值,hFgl2蛋白表达高时,平均灰度值数值越低。即:hFgl2蛋白表达情况与所测定的灰度值呈负向关系。以SSPS16.0统计软件分析比较各组中hFgl2蛋白的表达差异,以及在慢性乙型肝炎患者组和慢性重型乙型肝炎组中其表达与ALT、AST、TBil相关关系。
结果:
1.hFgl2蛋白在六组外周血单核细胞中表达的平均灰度值分别为142.20±1.29(A组),147.43±2.64(B组),140.04±2.90(C组),135.22±10.36(D组),135.11±2.10(E组),136.30±11.70(F组)。hFgl2蛋白在外周血单核细胞的MIAS医学分析图像中,慢性乙型肝炎轻中重度各组与慢性重型乙型肝炎组比较差异有统计学意义(P<0.05),即慢性重型乙型肝炎组的平均灰度值低于慢性乙型肝炎轻中重度各组的平均灰度值,提示慢性重型乙型肝炎组外周血单核细胞中hFgl2蛋白表达高于慢性乙型肝炎轻中重度各组。
2.hFgl2蛋白在慢性乙型肝炎组(B组,C组,D组)和肝癌组(F组)四组肝组织肝细胞中表达的平均灰度值分别为182.38±0.99(B组),181.84±2.64(C组),174.14±1.39(D组),177.30±6.09(F组)。hFgl2蛋白在肝组织表达的MIAS医学分析图像中,慢性乙型肝炎中度组、重度组两两比较,差异均有统计学意义(P<0.05),即慢性乙型肝炎重度组的平均灰度值低于慢性乙型肝炎轻中度组的平均灰度值,提示慢性乙型肝炎重度组肝细胞中hFgl2蛋白表达高于慢性乙型肝炎轻中度组。
3.在外周血单核细胞中,慢性乙型肝炎重度组hFgl2蛋白表达的平均灰度值与ALT和AST呈负相关(rs分别为-0.83和-0.84,P<0.05),即hFgl2蛋白的平均灰度值与慢性乙型肝炎重度组的肝细胞损伤程度呈负相关,即慢性乙型肝炎重度组外周血单核细胞中hFgl2蛋白表达强度与ALT、AST成正相关。hFgl2蛋白的平均灰度值与TBil的相关性经检验差异无统计学意义(P>0.05)。
4.hFgl2蛋白在肝癌组外周血单核细胞中平均灰度值为136.3±11.70(F组),在组织中平均灰度值为177.3±6.09(F组)。肝癌组与慢性乙型肝炎各组外周血单核细胞中hFgl2蛋白的平均灰度值比较差异均有统计学意义,即外周血单核细胞中hFgl2蛋白在肝癌组中表达高于慢性肝炎组;在组织中,hFgl2蛋白的平均灰度值与慢性乙型肝炎轻中度组的平均灰度值比较差异均有统计学意义,即hFgl2蛋白在肝癌组肝细胞中表达强于慢性乙型肝炎轻中度组,但与慢性乙型肝炎重度组与比较差异无统计学意义。
结论:
hFgl2蛋白能够表达于人外周血单核细胞中及慢性乙型肝炎重度患者和肝癌患者肝组织中。且hFgl2蛋白随着慢性肝炎病情严重程度加重其表达升高,而肝癌患者中hFgl2蛋白的表达强于慢性乙型肝炎轻中度患者。但组织和单核细胞中hFgl2蛋白的表达呈现不同步性。hFgl2凝血酶原酶有望成为慢性乙型肝炎重度患者肝细胞损伤程度的另一评价指标。
Objective: To detect the protein expressions of hfg12 (human fibrinogen like protein 2) from peripheral blood monocytes as well as liver tissues in patients with different degrees of chronic hepatitis B and liver cancer and try to make clear the correlation and possible involvement between these diseases and the expression of hFgl2.Research hFgl2 prothrombin enzyme positive expression of mononuclear cells and liver total bilirubin (TBil), alanine amino shift enzyme (ALT), the usable acid amino converting enzyme (AST) to determine the correlation between the degree of necrosis of liver cells, and understand the sensitivity of the reflection hFgl2 prothrombin enzyme expression determination of the clinical significance.
Methods:
1.Acquisition with complete clinical data of 37 cases with chronic hepatitis B, 20 cases of chronic fulminant hepatitis B (group E) and 21 cases of liver cancer (F) peripheral blood smear as experimental, 20 cases of the healthy check-up peripheral blood smear as control group (A);37 cases of chronic hepatitis B according to the guideline of prevention and treatment for chronic hepatitis B (2000 version) [1] is divided into mild (B), medium (C) or severe (D) three groups, mild in 15 cases, moderate 13 cases, severe 9 cases.
2.Collected the above 37 cases of chronic hepatitis B with liver biopsy tissues and 21 cases of liver cancer patients surgical resection of the liver tissue.
3.Application immunohistochemistry methods were used in the treatment of chronic hepatitis B group, detect Fgl2 protein expression of chronic severe hepatitis B group, liver group and the normal group peripheral blood monocytes.And using the two-step immunohistochemistry methods detected to Fgl2 protein expression in the collected in the liver tissue.
4.The statistical data of expression was analysed by SPSS16.0.
Results:
1.Fgl2 proteins was detected in six groups of peripheral blood monocytes.Its expression of average gray level respectively was(Group A):142.20±1.29,(Group B):147.43±2.64,(Group C):140.04±2.90,(Group D):135.22±10.36,(Group E):135.11±2.10,(Group F):136.30±11. 70.In MIAS medical analysis image,the average gray level of Fgl2 protein in peripheral blood monocytes, the expression of chronic hepatitis B group and chronic fulminant hepatitis group compared ,differences was statistically significant (P < 0.05).The normal group with the different degree chronic hepatitis B groups Fgl2 protein expression comparative difference was no statistically significant (P > 0.05).
2.Fgl2 proteins was detected in four groups of liver tissues.Its expression of average gray level respectively was(Group B):182.38±0.99,(Group C):181.84±2.64,(Group D):174.14±1.39. In MIAS medical analysis image,the average gray level of Fgl2 proteins in the liver tissue expression, chronic hepatitis B moderate group and severe group binary comparison, differences was statistically significant difference (P < 0.05).The expression of Fgl2 proteins in chronic hepatitis B light and moderate group,differences was not statistically significant (P > 0.05).
3.In peripheral blood monocytes,the expression of Fgl2 proteins average gray level correlates with the level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST).(rs respectively was -0.83 and -0.84 ,P<0.05).This indecates that Fgl2 protein expression with chronic hepatitis B severely group of severity were positively correlated.The average gray level of Fgl2 proteins by inspection TBil correlation was not statistically significant difference (P > 0.05).
4.In liver cancer group(F group),the average gray level of Fgl2 proteins was respectively 136.3±11.70 and 177.30±6.09 in peripheral blood monocytes and tissue.In peripheral blood monocytes in F group,the average gray level of Fgl2 proteins was different of from A group to E group,differences was statistically significant difference (P< 0.05).But in liver tissue, the average gray level of Fgl2 proteins F group was similiar to the chronic hepatitis B severe groups group(D group).
Conclusions:
hFgl2 protein can be in the human peripheral blood monocytes and in the liver tissue of in Chronic hepatitis B severe group and liver cancer group.hFgl2 protein expression could be high with the severity degree of chronic liver disease.The expression of hFgl2 prothrombin enzyme in liver cancer group was higher than chronic hepatitis B groups in PBMC.About liver tissue,the expression of Fgl2 prothrombin enzyme was similiar to the chronic hepatitis B severe group.hFgl2 protein expression in organizations and monocytes presented non-synchronous.hFgl2 prothrombin may become monitoring indicators of the liver cell damage degree in Chronic hepatitis B severe group.
引文
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