Department of Geratology, Yantai Yuhuangding Hospital, Yantai, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):3005-3011. doi: 10.26355/eurrev_201904_17582.
To explore the roles of interleukin-10 (IL-10), proNGF and p75NTR in apoptosis of brain tissues induced by intracerebral hemorrhage (ICH).
According to the time of sample collection after ICH, brain tissue samples were divided into < 6 h group, 6-24 h group (including 24 h), 24-72 h group (including 72 h) and > 72 h group. Meanwhile, 10 tissues that dropped from the beginning at the cortical stoma (distal part of the hematoma) were harvested as controls. AI in brain tissues around the hematoma after ICH was calculated based on TUNEL staining. Expression levels of IL-10, proNGF and p75NTR in brain tissues were determined by quantitative Real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Protein expressions of Bcl-2 and Bax were detected by Western blot. Rat cortical astrocytes were harvested and cultured in vitro. After transfection of IL-10 overexpression plasmid, expression levels of IL-10, proNGF and p75NTR were detected by Western blot.
AI increased in 6-24 h group, 24-72 h group and > 72 h group compared with < 6 h group and control group, which achieved the peak at 24-72 h. However, no significant difference in AI was observed between < 6 h group and control group. With the prolongation of ICH, IL-10 level gradually decreased and achieved the lowest level at 24-72 h. After 72 h, IL-10 level began to increase. Additionally, mRNA and protein levels of proNGF and p75NTR started to upregulate within 6 h of ICH, achieveing the peak at 24-72 h. Bcl-2 level gradually decreased after 6 h of ICH, while Bax level increased. We did not found significant difference in mRNA and protein levels of IL-10 in brain tissues around hematoma between < 6 h group and control group. With the prolongation of ICH, IL-10 level gradually decreased and achieved the lowest level at 24-72 h. After 72 h, IL-10 level began to increase. Transfection with IL-10 overexpression plasmid in rat astrocytes markedly downregulated protein levels of proNGF and p75NTR compared with those of controls.
IL-10 expression is downregulated in brain tissues around the hematoma after ICH. IL-10 alleviates inflammation and apoptosis by inhibiting levels of proNGF, p75NTR and Bax/Bcl-2, thus protecting brain tissue after ICH.
探讨白细胞介素 10(IL-10)、前神经生长因子(proNGF)和 p75NTR 在脑出血(ICH)诱导的脑组织细胞凋亡中的作用。
根据 ICH 后样本采集时间,将脑组织样本分为<6 h 组、6-24 h 组(包括 24 h)、24-72 h 组(包括 72 h)和>72 h 组。同时,从皮质造口(血肿远端)起始处采集 10 个组织作为对照。根据 TUNEL 染色计算 ICH 后血肿周围的 AI。通过定量实时聚合酶链反应(qRT-PCR)和 Western blot 分别测定脑组织中 IL-10、proNGF 和 p75NTR 的表达水平。通过 Western blot 检测 Bcl-2 和 Bax 的蛋白表达。采集大鼠皮质星形胶质细胞并进行体外培养。转染 IL-10 过表达质粒后,通过 Western blot 检测 IL-10、proNGF 和 p75NTR 的表达水平。
与<6 h 组和对照组相比,6-24 h 组、24-72 h 组和>72 h 组的 AI 在 6-24 h 时增加,在 24-72 h 时达到峰值,但<6 h 组与对照组之间的 AI 无显著差异。随着 ICH 的延长,IL-10 水平逐渐降低,并在 24-72 h 时达到最低水平。72 h 后,IL-10 水平开始增加。ICH 后 6 h 内,proNGF 和 p75NTR 的 mRNA 和蛋白水平开始上调,在 24-72 h 时达到峰值。ICH 后 6 h,Bcl-2 水平逐渐降低,而 Bax 水平升高。我们没有发现<6 h 组和对照组血肿周围脑组织中 IL-10 的 mRNA 和蛋白水平有显著差异。随着 ICH 的延长,IL-10 水平逐渐降低,并在 24-72 h 时达到最低水平。72 h 后,IL-10 水平开始增加。转染大鼠星形胶质细胞的 IL-10 过表达质粒后,与对照组相比,proNGF 和 p75NTR 的蛋白水平明显下调。
ICH 后血肿周围脑组织中 IL-10 表达下调。IL-10 通过抑制 proNGF、p75NTR 和 Bax/Bcl-2 的水平来减轻炎症和细胞凋亡,从而保护 ICH 后的脑组织。