Zhu Huizhi, Wang Kun, Yang Lei, Xu Qingwen, Ren Fengchun, Liu Xiangguo
Department of Respiratory Medicine, First Affiliated Hospital of Anhui University of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230031, China.
Graduate School, Anhui University of Chinese Medicine, Hefei 230031, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Dec 30;40(12):1703-1711. doi: 10.12122/j.issn.1673-4254.2020.12.02.
To observe the effect of (YHPC) granule on miR-139-5p, Notch1/Hes1 pathway and homing of bone marrow-derived mesenchymal stem cells (BMSCs) in asthmatic rats.
Fifty SD rats were randomized divided into normal control (NC) group, asthmatic model group, BMSCs transplantation group, BMSCs + dexamethasone (0.0625 mg/kg daily) group, and BMSCs+YHPC granule (3.5 g/kg daily) group. In all but the normal control group, asthmatic rat models were established by ovalbumin challenge, and BMSCs (1×10/mL) transplantation the tail vein was performed in the latter 3 groups on last day of ovalbumin challenge. In all the groups, lung pathologies of the rats were evaluated using HE staining after the treatments. Flow cytometry was employed to detect pulmonary expression of CXCR4 protein, and ELISA was used to determine the expressions of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in the lung tissue. The expressions of CXCR4, Notch1 and Hes1 in bronchial epithelial cells was examined using immunofluorescence assay. RT-PCR was used to detect the expressions of miR-139-5p, Notch1, Jagged1, RBP-J and Hes1 mRNAs, and the protein expressions of Notch1, Jagged1 and Hes1 were detected with Western blotting.
Compared with the normal control rats, the asthmatic rats exhibited significantly increased expressions of CXCR4, IL-4, Notch1, Jagged1, RBP-J, and Hes1 mRNA and Notch1, Jagged1, and Hes1 proteins and lowered expressions of INF-γ mRNA and miR-139-5p in the lung tissues ( < 0.05 or 0.01). Compared with those in the asthmatic model group, the mRNA expressions of CXCR4, IFN-γ, and miR-139-5p increased and the expressions of IL-4, Notch1, Jagged1, RBP-J, and Hes1 mRNA and Notch1, Jagged1, and Hes1 proteins decreased significantly in the 3 groups with BMSCs transplantation ( < 0.05 or 0.01). The rats in BMSCs+YHPC granule group showed significantly higher CXCR4, IFN-γ, and miR-139-5p mRNA expressions and lower IL-4 and Notch1 mRNA expressions than those in BMSCs transplantation group ( < 0.05).
YHPC granule can enhance the inhibitory effect of BMSCs homing on Th2 inflammatory response in asthmatic rats by up-regulating miR-139-5p and down-regulating Notch1/Hes1 pathway.
观察（药名未给出，暂称YHPC）颗粒对哮喘大鼠骨髓间充质干细胞（BMSCs）的miR-139-5p、Notch1/Hes1通路及归巢的影响。
将50只SD大鼠随机分为正常对照组（NC组）、哮喘模型组、BMSCs移植组、BMSCs+地塞米松（每日0.0625mg/kg）组和BMSCs+YHPC颗粒（每日3.5g/kg）组。除正常对照组外，其余各组通过卵清蛋白激发建立哮喘大鼠模型，后3组在卵清蛋白激发的最后一天经尾静脉进行BMSCs（1×10/mL）移植。处理后，采用HE染色评估各组大鼠的肺组织病理变化。运用流式细胞术检测肺组织中CXCR4蛋白的表达，采用ELISA法测定肺组织中干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）的表达。采用免疫荧光法检测支气管上皮细胞中CXCR4、Notch1和Hes1的表达。运用RT-PCR检测miR-139-5p、Notch1、Jagged1、RBP-J和Hes1 mRNA的表达，采用蛋白质印迹法检测Notch1、Jagged1和Hes1的蛋白表达。
与正常对照大鼠相比，哮喘大鼠肺组织中CXCR4、IL-4、Notch1、Jagged1、RBP-J和Hes1 mRNA以及Notch1、Jagged1和Hes1蛋白的表达显著升高，而INF-γ mRNA和miR-139-5p的表达降低（P<0.05或0.01）。与哮喘模型组相比，BMSCs移植的3组中CXCR4、IFN-γ和miR-139-5p的mRNA表达升高，IL-4、Notch1、Jagged1、RBP-J和Hes1 mRNA以及Notch1、Jagged1和Hes1蛋白的表达显著降低（P<0.05或0.01）。BMSCs+YHPC颗粒组大鼠的CXCR4、IFN-γ和miR-139-5p mRNA表达显著高于BMSCs移植组（P<0.05），IL-4和Notch1 mRNA表达低于BMSCs移植组。
YHPC颗粒可通过上调miR-139-5p和下调Notch1/Hes1通路，增强BMSCs归巢对哮喘大鼠Th2炎症反应的抑制作用。
