【摘要】 目的 探讨茶多酚对盲肠结扎穿孔术(CLP)所致大鼠脓毒症急性肾损伤(AKI)的保护作用。方法 130只雄性SD大鼠随机分为:对照组(10只大鼠),假CLP组(假手术组,40只大鼠),脓毒症组(CLP组,40只大鼠),茶多酚组(40只大鼠)。采用CLP复制脓毒症模型。茶多酚组在造模后即予茶多酚400 mg/(kg·d)灌胃。假手术组、CLP组和茶多酚组按腹壁缝合后2、12、24、48 h时点分成4个亚组(每组8只大鼠),分别在各时点随机处死8只大鼠,采用酶联免疫(ELISA)法分析测定血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、诱导型一氧化氮合酶(iNOS)、高迁移率族蛋白B-1(HMGB-1)的水平;同时检测血清肌酐(Cr)、尿素氮(BUN)的水平;免疫组化法检测大鼠肾组织核转录因子(NF-KB)蛋白表达;采用光镜和透射电镜观察肾组织病理变化。结果 与脓毒症组比较[Cr 12 h:(57.6±3.45)μmol/L,24 h:(79.4±5.79)μmol/L,48 h:(78.4±5.52)μmol/L;BUN 12 h:(11.82±3.83)mmol/L,24 h:(23.09±3.75)mmol/L,48 h:(20.85±3.54)mmol/L],茶多酚组大鼠的肾功能[Cr 12 h:(52.8±3.92)μmol/L,24 h:(65.9±4.13)μmol/L,48 h:(60.8±5.97)μmol/L;BUN 12 h:(8.47±2.58)mmol/L,24 h:(15.20±3.69)mmol/L,48 h:(13.40±3.41)mmol/L]明显改善,差异有统计学意义(P<0.05)。与脓毒症组比较[TNF-α2 h:(25.70±3.67)μg/mL,12 h:(65.16±4.57)pg/mL,24h:(78.95±7.83)pg/mL,48 h:(74.71±6.92)pg/mL;IL-10 2 h:(28.70±3.67)μg/L,12 h:(74.16±4.57)μg/L,24 h:(96.95±7.83)μg/L,48 h:(85.71±5.72)μg/L;iNOS 2 h:(46.58±4.72)U/L,12 h:(64.31±6.28)U/L,24 h:(59.05±7.54)U/L,48 h:(47.83±6.08)U/L;HMGB-1 12 h:(27.83±6.38)ng/mL,24 h:(64.31±8.28)ng/mL,48 h:(49.05±8.54)ng/mL],茶多酚组大鼠的炎症反应水平[TNF-α2 h:(20.56±3.29)pg/mL,12 h:(43.62±5.83)pg/mL,24 h:(56.32±7.56)pg/mL,48 h:(50.96±7.28)pg/mL;IL-10 2 h:(25.39±3.29)μg/L,12 h:(58.72±6.83)μg/L,24 h:(71.94±7.52)μg/L,48 h:(62.36±7.24)μg/L;iNOS 2 h:(35.49±3.96)U/L,12 h:(54.87±7.25)U/L,24h:(48.23±6.24)U/L,48 h:(37.16±7.51)U/L;HMGB-1 12 h:(24.57±7.25)ng/mL,24 h:(38.23±6.24)ng/mL,48 h:(37.16±7.51)ng/mL]明显下降,差异有统计学意义(P<0.05)。与脓毒症组比较(18 709.382±1550.263),茶多酚组大鼠的肾组织NF-κB表达水平(9274.416±1356.397)明显降低,差异有统计学意义(P<0.01)。脓毒症组大鼠肾脏病理损伤严重,茶多酚组大鼠肾脏的病理改变明显减轻。结论茶多酚能够抑制炎症反应,进而改善肾功能,对脓毒症大鼠AKI具有一定的保护作用。更多还原

【Abstract】 Objective To investigate the protective effect of tea polyphenol on caecal ligation and puncture( CLP)-induced acute kidney injury( AKI). Methods 130 healthy male SpragueDawley rats(250 ～300 g) were randomly divided into 4 groups: normal control group,Sham group,CLP model group( sepsis group), tea polyphenol group. The rat models of sepsis were established by caecal ligation and puncture. In treatment group, rats were infused with tea polyphenols once a day with a dose of 400 mg/kg body weight after CLP surgery. Sham group, CLP model group and tea polyphenol group were divided into 2, 12, 24, 48 h subgroups with 10 rats in each. Rats were sacrificed and the tissue samples including kidney and blood samples were collected. The kidney function, plasma cytokines interleukin-10( IL-10) and tumor necrosis factor-alpha(TNF-α), inducible nitric oxide synthase(iNOS) and high mobility group boxlprotein(HMGB-1) were analyzed. Cytokines were determined by the ELISA method. The expression of nuclear factor-kappaB(NF-κB) protein in kidney was detected by immnunohistochemistry method. Pathological changes of kidney tissues were observed by light and transmission electron microscopy. Results Compared with the sepsis group [ Cr(μmol/L) : 12 h:(57.6 ±3.45), 24 h:(79.4 ±5.79), 48 h:(78.4 ±5.52); BUN(mmol/L):12 h:(11. 82 ±3. 83),24 h:(23.09 ±3.75),48 h:(20. 85 ±3.54) ],the renal function of the tea polyphenol group rats [Cr(μmol/L): 12 h:(52. 8 ±3. 92),24 h:(65. 9 ±4. 13),48 h:(60. 8 ±5.97); BUN(mmol/L): 12 h:(8.47 ±2.58), 24 h:(15.20 ±3.69), 48 h:(13.40 ±3.41) ] was significantly improved, and the differences are significantly(P < 0.05). Compared with the sepsis group[TNF-α(pg/mL): 2 h:(25.70±3.67), 12 h:(65.16±4.57),24 h:(78.95±7.83),48 h:(74.71 ±6.92); IL-10(μg/L) : 2 h:(28. 70 ±3. 67), 12 h:(74.16 ±4.57), 24 h:(96.95 ±7.83),48 h:(85.71 ±5.72); iNOS(U/L) : 2 h:(46.58 ±4.72),12 h:(64.31 ±6.28),24 h:(59.05 ±7.54),48 h:(47.83 ±6.08); HMGB-1(ng/mL): 12 h:(27.83 ±6.08),24 h:(49.05 ±8.54),48 h:(64.31 ±8.28) ],the levels of inflammation in the tea polyphenol group rats[TNF-α(pg/mL):2 h:(20.56±3.29),12 h:(43.62±5.83),24 h:(56.32±7.56),48 h:(50.96±7.28);IL-10(μg/L) 2 h:(25. 39 ±3.29),12 h:(58.72 ±6.83),24 h:(71.94±7.52),48 h:(62.36±7.24)); iNOS(U/L) 2 h:(35.49±3.96),12 h:(54.87±7.25),24 h:(48.23 ± 6.24), 48 h:(37. 16±7.51); HMGB-1(ng/mL): 12 h:(24.57 ±7.25), 24 h:(38.23 ± 6. 24),48 h:(37. 16 ± 7. 51)] were significantly reduced,and the differences are significantly(P <0. 05). Compared with the sepsis group(18 709. 382 ± 1550. 263), the expression level of NF-κB was significantly decreased in the tea polyphenol group rats(9274.416 ± 1356. 397),and the difference was significant(P < 0. 01). Renal pathological damage was severe in sepsis group,but tea polyphenol lessened the kidney histological changes. Conclusion Tea polyphenol can inhibit the levels of TNF-α, IL-10, iNOS and HMGB-1 in serum, thus modify the inflammatory response and provided protective effects against acute kidney injury induced by sepsis in rats.更多还原