Neuroprotective effect of Naochuxue prescription on intracerebral hemorrhage: inhibition of autophagy downregulating high mobility group box-1.

Jin Hong, Wang Xinna, Wang Ruonan, L I Jinjian, Y U Junchao, Zhao Dexi, Zhai Lu
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Abstract

Objective: To determine the molecular mechanisms underlying the neuroprotective effects of Naochuxue prescription (,NCXP) in rats with intracerebral hemorrhage (ICH).

Methods: Sprague-Dawley rats were injected with collagenase to generate ICH models, which were then randomly divided into six groups, including control, sham, model, and three intervention groups. The intervention groups received different doses of NCXP (0.13, 0.26, and 0.52 g/kg) daily for 10 d. High-performance liquid chromatography (HPLC) was used to analyze the chemical characteristics of NCXP. The neurobehavioral outcomes of the rats were evaluated using neurological deficit scores (Zea Longa 5) and the corner turn test. Pathomorphological changes in perihematomal tissues after ICH were observed using hematoxylin and eosin staining. Immunohistochemistry (IHC) was used to detect the inflammation expression of interleukin 6 (IL-6) and toll-like receptor 4 (TLR4). High mobility group box-1 (HMGB1), Beclin1, microtubule-associated protein 1 light chain 3 beta (LC3), and sequestosome 1 (p62) were detected using real-time quantitative polymerase chain reaction and Western blotting in perihematomal tissues.

Results: HPLC showed that the NCXP had good stability. Rats with ICH had severe neurological function deficits compared to the control group. IHC results showed that NCXP significantly downregulated the expression of the inflammatory proteins IL-6 and TLR4. ICH rats treated with NCXP showed less neurological injury than the model group, accompanied by a significantly decreased expression of HMGB1, Beclin1, and LC3 and an increased expression of p62.

Conclusions: The neuroprotective effect of NCXP alleviated inflammation and autophagy possibly by downregulating HMGB1 expression. However, further research on the signaling pathways is required to verify this hypothesis.

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野菊花方对脑出血的神经保护作用:抑制自噬下调高迁移率基团框-1
目的方法:向Sprague-Dawley大鼠注射胶原酶,制成ICH模型,然后随机分为6组,包括对照组、假组、模型组和3个干预组:方法:给Sprague-Dawley大鼠注射胶原酶以产生ICH模型,然后将其随机分为六组,包括对照组、假组、模型组和三个干预组。采用高效液相色谱法(HPLC)分析 NCXP 的化学特性。使用神经功能缺损评分(Zea Longa 5)和转角测试评估大鼠的神经行为结果。使用苏木精和伊红染色观察 ICH 后血肿周围组织的病理形态学变化。免疫组化(IHC)用于检测白细胞介素6(IL-6)和类收费受体4(TLR4)的炎症表达。采用实时定量聚合酶链反应和 Western 印迹法检测血细胞周围组织中的高迁移率基团盒-1(HMGB1)、Beclin1、微管相关蛋白 1 轻链 3 beta(LC3)和序列体 1(p62):高效液相色谱法显示,NCXP 具有良好的稳定性。与对照组相比,ICH 大鼠有严重的神经功能缺损。IHC 结果显示,NCXP 能显著降低炎症蛋白 IL-6 和 TLR4 的表达。与模型组相比,接受 NCXP 治疗的 ICH 大鼠神经损伤较轻,同时 HMGB1、Beclin1 和 LC3 的表达明显减少,p62 的表达增加:NCXP的神经保护作用可能通过下调HMGB1的表达缓解了炎症和自噬。然而,要验证这一假设,还需要进一步研究信号通路。
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