健脾活血方对Lieber-DeCarli酒精性脂肪肝大鼠肠道菌群的影响

作者	单位
汪海慧	上海高校中医内科学E-研究院
平键	肝肾疾病病证教育部重点实验室
彭景华	上海高校中医内科学E-研究院
胡义扬	上海中医药大学附属曙光医院肝病研究所
成扬	上海中医药大学附属曙光医院肝病研究所
陈高峰	上海高校中医内科学E-研究院

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中文摘要:

目的研究健脾活血方对Lieber-DeCarli酒精性脂肪肝大鼠肠道菌群的影响。方法 40只SD大鼠随机分为正常组、无酒精液体饲料组、酒精液体饲料组和健脾活血方干预组,每组10只。干预组按照1.0mL/100g每天1次灌胃给予健脾活血方,其他组灌服等量生理盐水,连续8周。实验结束处死大鼠,采集血标本和肝脏组织,检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)活性以及门静脉血浆内毒素LPS水平。肝组织行HE染色和油红O染色检测肝脏病理改变。收集大鼠粪便,Bead-beating法从大鼠粪便中抽提肠道菌群总DNA,进行ERIC-PCR指纹图谱检测,对图谱作相似性聚类分析。结果与正常组(U/L,ALT:31.15±7.04,AST:53.23±10.28)及无酒精饲料组(ALT:26.96±8.12,AST:52.09±8.62)比较,酒精饲料组ALT(92.72±25.83),AST(72.60±23.31)显著升高(P<0.01),干预组ALT(65.28±20.36)和AST(59.11±10.32)活性均显著下降(P<0.01,P<0.05)。病毒学检查显示酒精饲料组大鼠肝组织内脂肪沉积明显,干预组显著减轻。正常组和无酒精饲料组大鼠门静脉LPS水平(EU/mL,0.033±0.010,0.043±0.018)极低,而酒精液体饲料组(0.541±0.085)显著升高(P<0.01),干预组显著降低LPS水平(0.349±0.098,P<0.01)。ERIC-PCR指纹图谱的聚类分析发现酒精液体饲料组大鼠肠道细菌群落发生了明显改变,而健脾活血方能够部分恢复肠道细菌群落的正常构成。结论健脾活血方对酒精性脂肪肝大鼠有良好的保护作用,并能在一定程度上调节肠道菌群的正常构成。

英文摘要:

Objective To study the effect of Jianpi Huoxue Recipe(JPHXR) on the gut flora in rats with alcoholic fatty liver (AFL) induced by Lieber-DeCarli liquid diet. Methods Forty Sprague-Dawley rats were divided into 4 groups:A:normal rats,B:rats fed with non-alcoholic liquid diet,C:rats fed with ethanol liquid diet to make AFL model,and D:AFL model rats intervened by gastrogavage of JPHXR 1.0 mL/100 g per day for 8 successive weeks,10 rats in each group. Except those in Group D (to them an equal volume normal saline was given for Successive instead),JPXHR was administered to rats in other three groups. At the end of experiment,rats were sacrificed,their blood and liver tissue samples were collected for determining serum activities of alanine transaminase (ALT) and aspartate aminotransferase (AST),endotoxin level in portal vein (expressed by lipopolysacchrides content,abbr. as LPS),and pathological examination of liver with HE staining and oil O red staining. Moreover,total DNA of gut flora were extracted from fresh rat fecal samples by Bead-beating method for determining the ERIC-PCR fingerprint,and a cluster analysis on the fingerprint was performed. Results Compared with the levels of ALT and AST in Group A (31.15±7.04 U/L,and 53.23±10.28 U/L respectively) and Group B (26.96±8.12 and 52.09±8.62),the corresponding levels in Group C (92.72±25.83 and 72.60±23.31) significantly increased (P<0.01),while the increments in Group D (65.28±20.36 and 59.11±10.32) were decroased(P<0.01,P<0.05). Pathological examination showed marked fat deposition in Group C,but which was significantly reduced in Group D. Endotoxin level in the portal vein was (0.033±0.010,EU/mL) in Group A and 0.043±0.018 in Group B,which was increased significantly in Group C (0.541±0.085,P<0.01) and Group D (0.349±0.098 EU/mL,P<0.01),but the increase in Group C was more significant (P<0.01). The cluster analysis of ERIC-PCR fingerprint showed significant changes in gut flora of Group C and D,which was in Group D partially recovered. Conclusions JPHXR had good preventive effect against alcoholic fatty liver in rats,and could modify the structure of gut flora to some extent.

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