目的 探討青黛對濕熱內(nèi)蘊(yùn)型潰瘍性結(jié)腸炎大鼠抗炎修復(fù)和免疫功能的影響,并探討其機(jī)制。方法 60只大鼠隨機(jī)分為6組,分別為對照組、模型組、青黛低劑量組、青黛中劑量組、青黛高劑量組以及柳氮磺胺吡啶(SASP)組、每組10只。青黛低、中、高劑量組分別ig 0.6、1.2、2.4 g/kg青黛懸濁液,SASP組ig 500 mg/kg柳氮磺吡啶懸濁液,對照組、模型組ig等體積生理鹽水。檢測各組大鼠血清白細(xì)胞介素(IL)-4、IL-10和腫瘤壞死因子-α(TNF-α)含量;流式細(xì)胞術(shù)檢測外周血單個核細(xì)胞(PBMC)中CD4⁺、CD8⁺T淋巴細(xì)胞百分率;放射免疫法檢測超氧化物歧化酶(SOD)和丙二醛(MDA)水平;熒光實時定量PCR法檢測蛋白激酶(ERK)、p38絲裂原活化蛋白激酶(p38 MAPK)和MEK mRNA相對表達(dá);免疫印跡法檢測ERK、p38 MAPK和MEK蛋白表達(dá)。結(jié)果 與對照組相比,模型組大鼠血清中IL-4、IL-10降低,TNF-α升高,PBMC中CD4⁺、CD8⁺ T淋巴細(xì)胞含量降低,大鼠結(jié)腸組織中ERK、p38 MAPK mRNA和蛋白表達(dá)升高、MEK mRNA和蛋白表達(dá)降低(P<0.05);與模型組相比,青黛低、中、高劑量組大鼠血清中IL-4、IL-10升高,TNF-α降低,PBMC中CD4⁺、CD8⁺ T淋巴細(xì)胞升高,ERK、p38 MAPK mRNA和蛋白表達(dá)降低,MEK mRNA和蛋白表達(dá)升高(P<0.05),青黛低、中、高劑量組間比較差異存在統(tǒng)計學(xué)意義(P<0.05)。對照組腸組織結(jié)構(gòu)完好,模型組結(jié)腸腸壁糜爛、潰瘍形成,炎癥明顯;青黛低劑量組仍伴隨較大潰瘍形成及炎癥浸潤;青黛中劑量組腸壁輕微增厚;青黛高劑量組與SASP組腸壁結(jié)構(gòu)尚完整,有極少量炎癥細(xì)胞浸潤。結(jié)論 青黛能夠降低濕熱內(nèi)蘊(yùn)型潰瘍性結(jié)腸炎大鼠機(jī)體內(nèi)炎癥因子的產(chǎn)生,抑制炎癥反應(yīng),促進(jìn)免疫功能的恢復(fù),其機(jī)制與調(diào)節(jié)ERK/p38 MAPK/MEK信號通路有關(guān)。

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

河北省中醫(yī)藥管理局科研計劃項目(2015012)