消痰化瘀中药对非酒精性脂肪肝模型大鼠PGC-1α mRNA表达及胰岛素抵抗的影响

Effects of phlegm-resolving and stasis-removing herbal drugs on PGC-1α mRNA expressions and insulin resistance in NAFLD rats

目的观察消痰化瘀中药对非酒精性脂肪肝(nonalcoholic fatty liverdisease, NAFLD)模型大鼠肝组织过氧化物酶体增殖激活受体(peroxisome proliferator-activated receptor, PPAR)γ辅助活化因子-1α (PPARγ coactivator-1α, PGC-1α)mRNA表达及胰岛素抵抗的影响,探讨其治疗NAFLD的作用机制。方法将60只SPF级SD大鼠按随机数字表法分为正常组、模型组、东宝肝泰组及消痰化瘀中药高、中、低剂量组,每组10只。通过饲养高脂饲料复制NAFLD大鼠模型。消痰化瘀中药高、中、低剂量组灌胃消痰化瘀药液43.34、32.50、21.67 g/kg,东宝肝泰组灌胃东宝肝泰片混悬液0.9 g/kg,正常组、模型组灌胃等体积蒸馏水,1次/d,连续8周。观察各组大鼠血清TC、TG、ALT、AST、FBG、游离脂肪酸(free fatty acid, FFA)、胰岛素(fasting insulin, FINS)、胰岛素抵抗指数(Insulin Resistance Index, HOMA-IR)和肝组织中TC、TG、FFA水平,采用PCR检测肝组织PGC-1α mRNA表达水平,采用HE染色观察肝组织病理形态学改变。结果与模型组比较,消瘀化痰中药高、中、低剂量组血清TG[(0.55±0.10)mmol/L、(0.58±0.09)mmol/L、(0.67±0.11)mmol/L比(1.18±0.15)mmol/L]、TC[(1.48±0.24)mmol/L、(1.69±0.27)mmol/L、(1.74±0.27)mmol/L比(3.29±0.26)mmol/L]、FFA[(251.08±48.18)μmol/L、(277.53±56.73)μmol/L、(291.82±48.67)μmol/L比(432.19±67.83)μmol/L]、ALT[(29.32±4.17)U/L、(31.26±4.74)U/L、(33.56±5.18)U/L比(47.21±8.67)U/L]、AST[(11.05±2.18)U/L、(12.15±2.67)U/L、(12.96±2.93)U/L比(19.43±3.68)U/L]、FBG[(5.68±1.22)mmol/L、(6.86±1.36)mmol/L、(7.94±1.82)mmol/L比(11.88±2.54)mmol/L]、FINS[(8.48±1.22)mmol/L、(9.55±1.95)mmol/L、(9.96±1.74)mmol/L比(12.96±2.67)mmol/L]水平降低,HOMA-IR[(1.91±0.26)、(2.91±0.65)、(3.52±0.58)比(6.89±1.21)]降低,肝组织中FFA[(242.19±35.13)μmol/L、(259.78±29.33)μmol/L、(277.62±34.29)μmol/L比(436.48±52.15)μmol/L]、TG[(23.65±3.28)mmol/L、(24.41±3.15)mmol/L、(25.37±3.59)mmol/L比(15.98±2.37)mmol/L]、TC[(7.15±0.82)mmol/L、(8.60±0.95)mmol/L、(8.86±1.04)mmol/L比(36.98±4.28)mmol/L]降低,肝组织PGC-1a mRNA[(1.24±0.06)、(1.02±0.07)、(0.99±0.08)比(0.43±0.06)]表达增高。结论消痰化瘀中药可调控PGC-1α基因表达,抑制糖异生、减少肝糖输出,纠正脂代谢紊乱,改善胰岛素抵抗,可能是其治疗NAFLD的作用机理。

Objective To explore the mechanism of phlegm-resolving and stasis- removing herbals on NAFLD by observing expressions of PGC1a mRNA and insulin resistance. Methods A total of 60 male SD rats were randomly divided into the normal group, model group, positive medication control group, high-dose, middle-dose and low-dose group. The rats were fed with high-fat forage for 8 weeks. The positive medication control group were gavaged with Dongbao-Gantai liquid (0.9 g/kg/day), the high-dose, middle-dose and low-dose group were gavaged with Xiaotan-Huayu liquid (43.34、32.50、21.67 g/kg/day), and normal group, model group were gavaged with equal volume of distilled water. The drugs were given by 1 ml/100 g and last for 8 weeks. The levels of TC, TG, FFA, ALT, AST, FBG, FINS, and HOMA-IR in serum, and levels of TC, TG, and PGC-1a mRNA and pathological morphological changes in hepatic tissue were observed after 8 weeks. Results The levels of TG (0.55 ± 0.10 mmol/L, 0.58 ± 0.09 mmol/L, 0.67 ± 0.11 mmol/L vs. 1.18 ± 0.15 mmol/L), TC (1.48 ± 0.24 mmol/L, 1.69 ± 0.27 mmol/L, 1.74 ± 0.27 mmol/L vs. 3.29 ± 0.26 mmol/L), FFA (251.08 ± 48.18 μmol/L, 277.53 ± 56.73 μmol/L, 291.82 ± 48.67 μmol/L vs. 432.19 ± 67.83 μmol/L), ALT (29.32 ± 4.17 U/L, 31.26 ± 4.74 U/L, 33.56 ± 5.18 U/L vs. 47.21 ± 8.67 U/L), AST (11.05 ± 2.18 U/L, 12.15 ± 2.67 U/L, 12.96 ± 2.93 U/L vs. 19.43 ± 3.68 U/L), FBG (5.68 ± 1.22 mmol/L, 6.86 ± 1.36 mmol/L, 7.94 ± 1.82 mmol/L vs. 11.88 ± 2.54 mmol/L), FINS (8.48 ± 1.22 mmol/L, 9.55 ± 1.95 mmol/L, 9.96 ± 1.74 mmol/L vs. 12.96 ± 2.67 mmol/L), HOMA-IR (1.91 ± 0.26, 2.91 ± 0.65, 3.52 ± 0.58 vs. 6.89 ± 1.21) in serum of high-dose, middle-dose and low-dose groups were decreased than model group. Levels of FFA (242.19 ± 35.13 μmol/L, 259.78 ± 29.33 μmol/L, 277.62 ± 34.29 μmol/L vs. 436.48 ± 52.15 μmol/L), TG (23.65 ± 3.28 mmol/L, 24.41 ± 3.15 mmol/L, 25.37 ± 3.59 mmol/L vs. 15.98 ± 2.37 mmol/L), TC (7.15 ± 0.82 mmol/L, 8.60 ± 0.95 mmol/L, 8.86 ± 1.04 mmol/L vs. 36.98 ± 4.28 mmol/L) were in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly lower than the model group. The levels of PGC-1a mRNA (1.24 ± 0.06, 1.02 ± 0.07, 0.99 ± 0.08 vs. 0.43 ± 0.06) in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly higher than model group. Conclusions The phlegm-resolving and stasis-removing herbals may improve lipid metabolism by regulating the expression of PGC-1α mRNA, inhibiting gluconeogenesis and liver sugar output, correcting disturbance of lipid metabolism and improving insulin resistance.