目的 探讨青黛对湿热内蕴型溃疡性结肠炎大鼠抗炎修复和免疫功能的影响,并探讨其机制。方法 60只大鼠随机分为6组,分别为对照组、模型组、青黛低剂量组、青黛中剂量组、青黛高剂量组以及柳氮磺胺吡啶(SASP)组、每组10只。青黛低、中、高剂量组分别ig 0.6、1.2、2.4 g/kg青黛悬浊液,SASP组ig 500 mg/kg柳氮磺吡啶悬浊液,对照组、模型组ig等体积生理盐水。检测各组大鼠血清白细胞介素(IL)-4、IL-10和肿瘤坏死因子-α(TNF-α)含量;流式细胞术检测外周血单个核细胞(PBMC)中CD4⁺、CD8⁺T淋巴细胞百分率;放射免疫法检测超氧化物歧化酶(SOD)和丙二醛(MDA)水平;荧光实时定量PCR法检测蛋白激酶(ERK)、p38丝裂原活化蛋白激酶(p38 MAPK)和MEK mRNA相对表达;免疫印迹法检测ERK、p38 MAPK和MEK蛋白表达。结果 与对照组相比,模型组大鼠血清中IL-4、IL-10降低,TNF-α升高,PBMC中CD4⁺、CD8⁺ T淋巴细胞含量降低,大鼠结肠组织中ERK、p38 MAPK mRNA和蛋白表达升高、MEK mRNA和蛋白表达降低(P<0.05);与模型组相比,青黛低、中、高剂量组大鼠血清中IL-4、IL-10升高,TNF-α降低,PBMC中CD4⁺、CD8⁺ T淋巴细胞升高,ERK、p38 MAPK mRNA和蛋白表达降低,MEK mRNA和蛋白表达升高(P<0.05),青黛低、中、高剂量组间比较差异存在统计学意义(P<0.05)。对照组肠组织结构完好,模型组结肠肠壁糜烂、溃疡形成,炎症明显;青黛低剂量组仍伴随较大溃疡形成及炎症浸润;青黛中剂量组肠壁轻微增厚;青黛高剂量组与SASP组肠壁结构尚完整,有极少量炎症细胞浸润。结论 青黛能够降低湿热内蕴型溃疡性结肠炎大鼠机体内炎症因子的产生,抑制炎症反应,促进免疫功能的恢复,其机制与调节ERK/p38 MAPK/MEK信号通路有关。

Objective To explore the effect of Indigo Naturalis on anti-inflammatory repair and immune function in rats with ulcerative colitis induced by damp-heat, and to explore its mechanism.Methods Sixty rats were randomly divided into control group(n = 10),model group(n = 10), low dose group(n = 10), middle dose group(n = 10), high dose group(n = 10) and sulfasalazine(SASP) group(n = 10). The model rats in the Indigo Naturalis low, middle and high dose groups were given 0.6, 1.2 and 2.4 g/kg, respectively. The model rats in the SASP group were gavaged according to 500 mg/kg sulfasalazine suspension, and the control group and model group were given the same volume of normal saline. The contents of IL-4, IL-10 and TNF-α in serum, the percentage of CD4⁺ and CD8⁺ T lymphocytes in peripheral blood mononuclear cells(PBMC) were detected by flow cytometry, the levels of MDA and SOD were detected by radioimmunoassay, the mRNA expressions of ERK, p38 MAPK and MEK genes were detected by Real-time PCR method,and the expressions of ERK, p38 MAPK and MEK proteins were detected by western blotting.Results Compared with the control group, the serum IL-4 and IL-10 of the model group decreased, the TNF-α increased, the content of CD4⁺ and CD8⁺ T lymphocytes in PBMC decreased, and the mRNA expression and protein expression of ERK and p38 MAPK gene and MEK decreased in the colon tissue of rats in the model group. Compared with the model group, IL-4 and IL-10 in serum increased, TNF-α decreased, CD4⁺ and CD8⁺ T lymphocytes in PBMC increased, mRNA expression and protein expression of ERK and p38 MAPK decreased and MEK increased in serum of rats in low, middle and high Indigo Naturalis groups. The intestinal tissue structure of the control group was intact, and the intestinal wall erosion, ulcer formation and inflammation of the model group were obvious; the low dose group was still accompanied by large ulcer formation and inflammatory infiltration; the middle dose group of indigo slightly thickened; the high dose group and SASP group intestinal wall structure was still intact, with a very small amount of inflammatory cell infiltration.Conclusion Indigo Naturalis can reduce the production of inflammatory factors, inhibit the inflammatory reaction and promote the recovery of immune function in rats with ulcerative colitis of damp-heat type, and its mechanism is related to the regulation of ERK/p38 MAPK/MEK signaling pathway.

R285.5

河北省中医药管理局科研计划项目(2015012)