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目的研究非酒精性脂肪性肝病(nonalcoholic fatty liver disease,NAFLD)兔脂肪肝程度与肝组织硫化氢(H2S)浓度的相关性。方法 40只日本大耳白兔按随机数字法分为重度NAFLD组(重度组)、轻度NAFLD组(轻度组)、对照组。重度组给予高脂饲料160g/d,轻度组给予高脂饲料80g/d+普通饲料80g/d,对照组给予普通饲料160g/d。饲养至10周开始,重度组每周行肝脏CT检测了解NAFLD程度,至13周重度组达到重度NAFLD,终止实验。饲养前耳中动脉采血5ml于肝素管中,离心后保存于-20℃的冰柜中待检。实验结束腹主动脉抽血5ml于肝素管中,离心后保存于-20℃的冰箱中待检。全部血浆按照设备、试剂操作常规检测实验前后甘油三酯(triglyceride,TG)、胆固醇(totol cholesteral,TC)。肝组织匀浆检测肝组织H2S浓度。胱硫醚-γ-裂解酶(cystathionine-γ-lyase,CSE)免疫组织化学染色、HE染色,光镜下观察肝脏组织形态学改变。结果①TC、TG:饲养前重度组、轻度组、对照组TC、TG比较无统计学差异(P>0.05)。饲养后重度组TC、TG分别为(32.12±1.25)、(6.02±2.12)mmol/L,轻度组TC、TG分别为(18.34±2.10)、(4.39±1.93)mmol/L,与饲养前比较有统计学差异(P<0.01)。饲养后重度组TG、TC高于轻度组(P<0.01)。②肝组织H2S浓度:重度组、轻度组肝组织H2S明显下降,重度组、轻度组与对照组比较均有统计学差异(P<0.01);重度组与轻度组比较也有统计学差异(P<0.05)。③肝脏病理学:重度组重度NAFLD,轻度组轻～中度NAFLD,对照组正常肝脏。轻度组CSE免疫组织化学染色CSE阳性细胞强于正常组,重度组强于轻度组。结论 NAFLD兔脂肪肝程度与肝组织H2S浓度负相关,可以通过脂肪肝程度负向推测肝组织H2S浓度。肝组织存在CSE表达代偿性增强。
Objective To investigate the correlation between the degree of fatty liver and the concentration of hydrogen sulfide (H2S) in nonalcoholic fatty liver disease (NAFLD) rabbits. Methods Forty Japanese white rabbits were divided into severe NAFLD group (mild group), mild NAFLD group (mild group) and control group according to random number method. Severe group given high fat diet 160g / d, mild group given high fat diet 80g / d + ordinary feed 80g / d, control group given general feed 160g / d. Feeding to the beginning of 10 weeks, the severe group every week liver CT scan to understand the extent of NAFLD to 13 weeks severe group reached severe NAFLD, termination of the experiment. Ear feeding ear artery 5ml blood heparin tube, centrifuged and stored in -20 ℃ freezer to be tested. The end of the experiment abdominal aorta blood 5ml in heparin tube, centrifuged and stored at -20 ℃ in the refrigerator to be tested. All plasma samples were tested for triglyceride (TG) and total cholesterol (TC) before and after the experiment according to equipment and reagents. Liver tissue homogenate detection of liver tissue H2S concentration. Cystathionine-γ-lyase (CSE) immunohistochemical staining, HE staining, light microscopic observation of liver tissue morphological changes. Results ①TC, TG: There was no significant difference in TC, TG between the control group and the severe group before feeding (P> 0.05). After rearing, the TC and TG were (32.12 ± 1.25) and (6.02 ± 2.12) mmol / L respectively in the severe group, while those in mild group were (18.34 ± 2.10 and 4.39 ± 1.93) mmol / L, There was a statistically significant difference (P <0.01). After rearing, TG and TC were higher in the severe group than in the mild group (P <0.01). (2) The concentration of H2S in liver tissue: The H2S level of liver tissue in severe group and mild group decreased significantly (P <0.01), there was significant difference between severe group and mild group (P <0.05). ③ liver pathology: severe group of severe NAFLD mild to moderate NAFLD group, the control group of normal liver. CSE immunohistochemical staining of mild CSE-positive cells was stronger than the normal group, severe group was stronger than mild group. Conclusions The level of fatty liver in NAFLD rabbit is negatively correlated with the concentration of H2S in liver tissue, and the concentration of H2S in liver tissue can be negatively estimated by the degree of fatty liver. There is compensatory enhancement of CSE expression in liver tissue.