沉香醇提物对哮喘小鼠的平喘作用及其机制研究

Anti-asthmatic effect of agarwood alcohol extract in mice and its mechanism

沉香记载有"纳气平喘"功效,但其作用机制不清且相关研究报道稀少。为了探究沉香的平喘及其作用机制,该研究采用卵清蛋白(ovalbumin, OVA)+氢氧化铝(Al(OH)3)激发联合OVA滴鼻诱导小鼠哮喘模型,考察通体结香技术产沉香(简称"通体沉香")醇提物的平喘作用及机制,比较不同结香方式所产沉香平喘作用差异。首先,通过评价哮喘小鼠的哮喘次数、观察肺组织病理损伤程度及计数外周血炎性细胞白细胞和嗜酸性粒细胞数,评价通体沉香的抗炎平喘作用;其次,通过酶联免疫(enzyme-linked immunoassay, ELISA)法检测小鼠血清中白细胞介素1β(interleukin^(-1)β,IL^(-1)β)、白细胞介素17(interleukin 17,IL^(-1)7)和白细胞介素10(interleukin^(-1)0,IL^(-1)0)的分泌水平,并通过实时定量PCR(Real-time quantitative-PCR, RT-PCR)检测组织中炎症及凋亡相关基因的表达,初步探究其平喘机制。结果显示,通体沉香醇提物能显著减少小鼠的哮喘次数、缓解哮喘小鼠病理损伤程度并改善外周血白细胞数和嗜酸性粒细胞数,降低炎性细胞因子IL^(-1)β和IL^(-1)7水平,升高抗炎细胞因子IL^(-1)0水平,同时下调IL^(-1)R、tumor necrosis factor receptor R (TNFR)、nuclear transcription factor-kappaB (NF-κB)、Bax及caspase-3 mRNA的表达,但对high mobility group box 1 protein B1(HMGB1)、caspase-8和Bcl-2的表达无明显影响,同等剂量下通体沉香醇提物作用优于野生沉香和火烙沉香醇提物。研究表明,通体沉香可显著抗炎平喘,其作用机制可能与抑制炎症和抗凋亡作用有关。

As recorded,agarwood has the function of improving qi reception and relieving asthma,but the underlying mechanism is unclear and rarely reported.Therefore,this study explored the anti-asthmatic effect of the alcohol extract of agarwood produced by the whole-tree agarwood-inducing technique(Agar-Wit)in the asthma mouse model induced by intraperitoneal injection of ovalbumin(OVA)+Al(OH)3 combined with intranasal administration of OVA and the mechanism,and compared the anti-asthmatic effects of agarwood induced with different methods.Firstly,the anti-inflammatory and anti-asthmatic effects of Agar-Wit agarwood in mice were evaluated based on the asthma frequency,lung tissue injury,and peripheral inflammatory white blood cell(WBC)count and eosinophil count.Then,the levels of interleukin^(-1)β(IL^(-1)β),IL^(-1)7,and IL^(-1)0 in serum of mice were detected by enzyme-linked immunoassay(ELISA)and the expression of inflammation-and apoptosis-related genes in tissues was measured by reverse transcription polyme-rase chain reaction(RT-PCR)so as to preliminarily explore the anti-asthmatic mechanism.Results showed that the alcohol extract of Agar-Wit agarwood significantly reduced asthma frequency,relieved pathological injury,improved peripheral WBC count and eosinophil count,decreased the levels of inflammatory cytokines IL^(-1)βand IL^(-1)7,elevated the level of anti-inflammatory cytokine IL^(-1)0,and down-regulated the mRNA expression of IL^(-1) R,tumor necrosis factor receptor R(TNFR),nuclear transcription factor-kappa B(NF-κB),Bax,and caspase 3,but had no significant influence on the expression of high-mobility group box 1(HMGB1)protein,caspase 8,and Bcl-2.The effect of Agar-Wit agarwood alcohol extract was better than that of wild agarwood alcohol extract and alcohol extract of agarwood induced with the burning-chisel-drilling method at the same dose.In conclusion,Agar-Wit agarwood can significantly alleviate inflammation and asthma,which is related to its anti-inflammation and anti-apoptosis activity.