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目的研究西罗莫司对非酒精性脂肪肝(NAFLD)大鼠调节性T细胞(Treg)/辅助性T细胞17(Th17)平衡的影响。方法 30只SD大鼠,按照体重随机分为3组:模型组、实验组、空白组,各10只。用高脂饮食制备NAFLD模型。造模成功后,实验组灌胃高脂饲料+西罗莫司1 mg·kg~(-1)·d~(-1),连续2周,空白组始终予基础饲料。用酶联免疫吸附法检测血清总胆固醇(TC)、三酰甘油(TG)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、白细胞介素(IL)-10、IL-17水平。以流式细胞仪检测外周血Treg细胞和Th17细胞比例。结果血清指标:空白组的TC、TG分别为(1.75±0.16),(0.84±0.05)mmol·L~(-1),空白组的ALT、AST(95.54±16.85),(38.9±11.87)U·L~(-1),空白组的IL-10、IL-17分别为(40.21±20.17),(6.23±1.02)pg·mL~(-1);模型组的TC、TG分别为(6.34±1.77),(1.29±0.13)mmol·L~(-1),模型组的ALT、AST分别为(355.38±24.33),(197.45±18.69)U·L~(-1),模型组的IL-10、IL-17分别为(73.26±45.34),(27.63±2.35)pg·mL~(-1)。肝指标:空白组的肝TC、TG分别为(0.82±0.05)(0.78±0.04)mmol·L~(-1);模型组的肝TC、TG水平分别为(1.26±0.15),(1.89±0.23)mmol·L~(-1),与空白组比较,模型组明显升高,差异有统计学意义(均P<0.05)。实验组上述指标:血清中,TC、TG分别为(4.65±1.63),(0.86±0.07)mmol·L~(-1),ALT、AST分别为(175.4±15.74),(85.25±12.26)U·L~(-1),IL-10、IL-17分别为(56.48±24.94),(11.04±1.62)pg·mL~(-1);肝中,TC、TG水平分别为(0.94±0.14),(1.21±0.15)mmol·L~(-1),与模型组比较,明显降低,差异均有统计学意义(均P<0.05)。模型组Treg/Th17比例为1.06±0.32,较空白组的3.12±0.65明显下降;实验组Treg/Th17比例为2.75±0.58,较模型组明显升高,组间差异均有统计学意义(均P<0.05)。结论高脂饮食诱导的NAFLD大鼠中,存在Treg/Th17失衡,西罗莫司可通过调节Treg/Th17的平衡而缓解NAFLD的炎症反应。
Objective To investigate the effect of sirolimus on the balance of regulatory T cells (Treg) / T helper 17 (Th17) in non-alcoholic fatty liver disease (NAFLD) rats. Methods Thirty SD rats were randomly divided into three groups according to body weight: model group, experimental group and blank group. NAFLD model was prepared by high fat diet. After successful modeling, the experimental group was given high-fat diet + sirolimus 1 mg · kg ~ (-1) · d ~ (-1) for 2 weeks, and the blank group was given basic feed. Serum total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin (IL) -10 and IL-17 levels were detected by enzyme-linked immunosorbent assay. Flow cytometry was used to detect the percentage of Treg cells and Th17 cells in peripheral blood. Results Serum levels of TC: TG in the blank group were (1.75 ± 0.16), (0.84 ± 0.05) mmol·L -1, respectively. The ALT, AST (95.54 ± 16.85), (38.9 ± 11.87) U · The levels of IL-10 and IL-17 in the blank group were (40.21 ± 20.17) and (6.23 ± 1.02) pg · mL -1, respectively. The TC and TG of the model group were (6.34 ± 1.77 and 1.29 ± 0.13 mmol·L -1 .The ALT and AST in the model group were (355.38 ± 24.33) and (197.45 ± 18.69) U · L -1 -10 and IL-17 were (73.26 ± 45.34) and (27.63 ± 2.35) pg · mL -1, respectively. Liver indexes: The levels of TC and TG in the blank group were (0.82 ± 0.05) and (0.78 ± 0.04) mmol·L -1, respectively. The levels of liver TC and TG in the model group were (1.26 ± 0.15) and (1.89 ± 0.23) mmol·L -1. Compared with the blank group, the model group was significantly increased, the difference was statistically significant (P <0.05). In the experimental group, the levels of TC and TG were (4.65 ± 1.63) and (0.86 ± 0.07) mmol·L -1, respectively, and the levels of ALT and AST were (175.4 ± 15.74) and (85.25 ± 12.26) U · The levels of L-1, IL-10 and IL-17 were (56.48 ± 24.94) and (11.04 ± 1.62) pg · mL -1, respectively. The levels of TC and TG were (0.94 ± 0.14 ), (1.21 ± 0.15) mmol·L -1, which were significantly lower than those in the model group (all P <0.05). The proportion of Treg / Th17 in the model group was 1.06 ± 0.32, which was significantly lower than that in the blank group (3.12 ± 0.65). The Treg / Th17 ratio in the experimental group was 2.75 ± 0.58, which was significantly higher than that in the model group <0.05). Conclusions Treg / Th17 imbalance exists in high fat diet induced NAFLD rats. Sirolimus can relieve the inflammatory response of NAFLD by regulating the balance of Treg / Th17.