不同中医证型非酒精性脂肪性肝病与外周血巨噬细胞表型的相关性研究

Correlation between different traditional Chinese medicine syndrome types and peripheral blood macrophage phenotype in non-alcoholic fatty liver disease

目的 探究非酒精性脂肪性肝病(NAFLD)患者中医证型与外周血巨噬细胞M1和M2表型的相关性。方法 选取2021年12月—2022年12月在黑龙江省中医医院肝胆脾胃二科门诊就诊并临床诊断为NAFLD的患者140例,根据中医辨证分型将患者分为湿浊内停组27例、肝郁脾虚组40例、湿热蕴结组32例、脾肾两虚组13例、痰瘀互结组28例。另从该院健康体检中心招募健康受试者15例作为健康受试组。检测各组受试者肝功能指标[血清谷丙转肽酶(ALT)、谷草转肽酶(AST)、γ-谷氨酰转肽酶(GGT)、碱性磷酸酶(ALP)]、血脂指标[三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)]、血清炎症因子指标[白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)和白细胞介素-4(IL-4)]、巨噬细胞表型、肝脏受控衰减参数(CAP),并经Pearson相关分析各组患者巨噬细胞表型与NAFLD病情的相关性。结果 与健康受试组相比,各中医证型组患者血清ALT、AST、GGT、ALP、TC、TG、LDL-C、IL-6、IL-8水平及M1型巨噬细胞百分比、M1/M2比值、CAP均显著增高(P均<0.05),血清HDL-C、IL-10、IL-4水平和M2型巨噬细胞百分比均显著降低(P均<0.05)。不同中医证型患者血清ALT、AST、GGT、ALP、TG、IL-6、IL-8水平及M1型巨噬细胞百分比、CAP由低到高分别为肝郁脾虚组、脾肾两虚组、湿浊内停组、痰瘀互结组、湿热蕴结组,血清HDL-C、IL-10、IL-4水平及M2型巨噬细胞百分比由高到低分别为肝郁脾虚组、脾肾两虚组、湿浊内停组、痰瘀互结组、湿热蕴结组,血清TC和LDL-C水平由低到高依次为肝郁脾虚组、湿浊内停组、湿热蕴结组、脾肾两虚组、痰瘀互结组。相关性分析发现,血清ALT、AST、GGT、ALP、TC、TG、LDL-C、IL-6、IL-8水平及CAP与M1型巨噬细胞百分比呈正相关(P均<0.05),与M2型巨噬细胞百分比呈负相关(P均<0.05);血清HDL-C、IL-10、IL-4水平与M1型巨噬细胞百分比呈负相关(P均<0.05),与M2型巨噬细胞百分比呈正相关(P均<0.05)。结论 NAFLD各中医证型中湿热蕴结型患者M1型巨噬细胞百分比最高,肝郁脾虚型患者M1型巨噬细胞百分比最低,而M2型巨噬细胞百分比规律与此相反,推测巨噬细胞极化参与NAFLD的发生和发展。

Objective It is to investigate the correlation between traditional Chinese medicine syndrome types and M1 and M2 phenotypes of peripheral blood macrophages in patients with nonalcoholic fatty liver disease(NAFLD).Methods One hundred and forty patients who were treated at the outpatient clinic of the Second Department of Liver,Gallbladder,Spleen and Stomach of Heilongjiang Provincial Hospital of Traditional Chinese Medicine from December 2021 to December 2022 and were clinically diagnosed with NAFLD were selected,and were divided into internal dampness and turbidity retention group(n=27),liver depressions and spleen deficiency group(n=40),dampness-heat accumulation group(n=32),spleen-kidney deficiency group(n=13),phlegm and stasis binding group(n=28).Another 15 healthy subjects were recruited from the hospital’s health checkup center as the healthy subject group.The liver function indexes[serum glutamate transpeptidase(ALT),glutamate transpeptidase(AST),gamma-glutamyl transpeptidase(GGT),alkaline phosphatase(ALP)],lipid indexes[triacylglycerol(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)],serum inflammation factor indexes[interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10(IL-10),and interleukin-4(IL-4)],macrophage phenotypes,hepatic controlled attenuation parameter(CAP)of subjects of each group were detected,and the correlation between macrophage phenotypes and NAFLD disease states in the patients of each group were analyzed by Pearson correlation analysis.Results Compared with the healthy subject group,the serum levels of ALT,AST,GGT,ALP,TC,TG,LDL-C,IL-6,IL-8 levels and the percentages of M1-type macrophages,M1/M2 ratio,and CAP were significantly increased,while the serum levels of HDL-C,IL-10,IL-4 and the percentages of M2-type macrophages were significantly decreased in the patients of each syndrome type group(all P<0.05).The serum levels of ALT,AST,GGT,ALP,TG,IL-6,IL-8 and percentage of M1-type macrophages and CAP of patients with different syndrome types from low to high were as follows,liver depressions and spleen deficiency group,spleen-kidney deficiency group,internal dampness and turbidity retention group,phlegm and stasis binding group,dampness-heat accumulation group,and the serum levels of HDL-C,IL-10,IL-4 and percentage of M2-type macrophages from high to low were as follows,liver depressions and spleen deficiency group,spleen-kidney deficiency group,internal dampness and turbidity retention group,phlegm and stasis binding group,dampness-heat accumulation group,and the serum levels of TC and LDL-C from low to high were as follows,liver depressions and spleen deficiency group,internal dampness and turbidity retention group,dampness-heat accumulation group,spleen-kidney deficiency group and phlegm and stasis binding group.Correlation analysis revealed that the serum levels of ALT,AST,GGT,ALP,TC,TG,LDL-C,IL-6,IL-8 and CAP were positively correlated with the percentage of M1-type macrophages(all P<0.05),and negatively correlated with the percentage of M2-type macrophages(all P<0.05);the serum levels of HDL-C,IL-10,IL-4 were negatively correlated with the percentage of M1-type macrophages(all P<0.05),and positively correlated with the percentage of M2 macrophages(all P<0.05).Conclusion Among the Chinese medicine syndrome types of NAFLD,the highest percentage of M1-type macrophages was found in patients with syndrome of damp-heat accumulation,the lowest percentage of M1-type macrophages was found in patients with syndrome of liver-depression and spleen-deficiency,and the rule of the percentage of M2-type macrophages was opposite to that of M1-type macrophages,which suggests that the macrophage polarization is involved in the occurrence and development of NAFLD.