外源性硫化氢通过Nrf2/ARE/HO-1通路对精神分裂症大鼠认知功能及肠道屏障功能的影响 #br#

目的 探讨外源性硫化氢（H2S）通过核因子E2相关因子2（Nrf2）/抗氧化反应元件（ARE）/血红素氧化酶1（HO-1）通路对精神分裂症大鼠认知功能及肠道屏障功能的影响。方法 40只SD大鼠按照随机数字表法分为对照组、模型组、H2S干预组（腹腔注射10 mg/kg的H2S供体GYY4137）、Nrf2抑制剂组（腹腔注射10 mg/kg的Nrf2抑制剂ATRA）、H2S+Nrf2抑制剂组（腹腔注射GYY4137和ATRA），每组8只。除对照组外，其余各组通过腹腔注射0.2 mg/kg的MK-801构建精神分裂症大鼠模型，模型构建成功后按照各组给药方式进行干预，模型组和对照组以生理盐水替代，连续干预7 d。Morris水迷宫实验测定大鼠的认知功能；HE染色观察大鼠海马组织及肠组织病理学变化；TUNEL检测大鼠肠黏膜细胞凋亡情况；酶联免疫吸附试验（ELISA）试剂盒测定大鼠血浆中H2S、D-乳酸含量及肠组织超氧化物歧化酶（SOD）、丙二醛（MDA）、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6水平；Western blot检测大鼠肠组织凋亡、Nrf2/ARE/HO-1通路相关蛋白表达情况。结果 与对照组相比，模型组大鼠海马组织、肠组织损伤加重，逃避潜伏期、血浆D-乳酸、肠黏膜细胞凋亡率、肠组织Caspase-3、Bax蛋白、MDA、TNF-α、IL-6、Nrf2、HO-1蛋白水平升高（P＜0.05），穿台次数、血浆H2S水平、小肠绒毛高度、隐窝深度、肠组织Bcl-2蛋白、SOD水平降低（P＜0.05）；与模型组相比，H2S干预组大鼠海马组织、肠组织损伤减弱，逃避潜伏期、血浆D-乳酸、肠黏膜细胞凋亡率、肠组织Caspase-3、Bax蛋白、MDA、TNF-α、IL-6水平降低（P＜0.05），穿台次数、血浆H2S水平、小肠绒毛高度、隐窝深度、肠组织Bcl-2蛋白、Nrf2、HO-1、SOD水平升高（P＜0.05）；Nrf2抑制剂可部分逆转H2S对模型大鼠认知功能、肠道屏障、海马组织的有益作用。结论 外源性H2S可明显改善精神分裂症大鼠的认知功能和肠道屏障功能，可能与激活Nrf2/ARE/HO-1通路有关。

The effects of exogenous hydrogen sulfide on cognitive function and intestinal barrier functionthrough Nrf2/ARE/HO-1 pathway in schizophrenic rats

Objective To investigate the effects of exogenous hydrogen sulfide (H2S) on cognitive function andintestinal barrier function through nuclear factor E2 related factor 2/antioxidant response element/heme oxidase 1 (Nrf2/ARE/HO-1) pathway in schizophrenic rats. Methods Forty SD rats were randomly divided into the control group, the modelgroup, the H2S intervention group (intraperitoneal injection of 10 mg/kg H2S donor GYY4137), the Nrf2 inhibitor group(intraperitoneal injection of 10 mg/kg Nrf2 inhibitor ATRA) and the H2S+Nrf2 inhibitor group (intraperitoneal injection ofGYY4137, ATRA), with eight in each group. Except for the control group, the other groups were injected intraperitoneallywith 0.2 mg/kg of MK-801 to construct the schizophrenic rat model. After the model was successfully constructed,intervention was carried out according to the administration method of each group. The model group and the control groupwere replaced with normal saline, continuous intervention for 7 days. Morris water maze test was used to measure thecognitive function of rats. Hematoxylin-eosin staining (HE) was used to observe the pathological changes of hippocampusand intestine tissue. TUNEL was used to detect the apoptosis of intestinal mucosal cells in rats. Enzyme-linkedimmunosorbent (ELISA) kit was used to determine the levels of H2S, D-lactic acid, intestinal tissue superoxide dismutase(SOD), MDA, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 in rat plasma. Western blot assay was used to detect the apoptosis of intestinal tissue and Nrf2/ARE/HO-1 pathway related proteins of rats. Results Compared with the controlgroup, the injury of hippocampal tissue and intestinal tissue were aggravated in the model group. The incubation period,plasma D-lactic acid, intestinal mucosal cell apoptosis rate, intestinal Caspase-3, Bax protein, MDA, TNF-α, IL-6, Nrf2and HO-1 were increased (P＜0.05). The number of puncture, the level of plasma H2S, the height of small intestinal villi, thedepth of recess, the level of Bcl-2 protein and SOD decreased in intestinal tissue (P＜0.05). Compared with the model group,the injury of hippocampus and intestine was alleviated in the H2S intervention group. The incubation period, plasma D-lacticacid, intestinal mucosal cell apoptosis rate, intestinal tissue Caspase-3, Bax protein, MDA, TNF-α and IL-6 decreased (P＜0.05). The number of puncture, the level of plasma H2S, the height of small intestinal villi, the depth of recess, the levels ofBcl-2, Nrf2, HO-1 protein and SOD in intestinal tissue increased (P＜0.05). Nrf2 inhibitor can partially reverse thebeneficial effects of H2S on cognitive function, intestinal barrier and hippocampus of model rats. Conclusion ExogenousH2S can significantly improve the cognitive function and intestinal barrier function of schizophrenic rats, which may beachieved by activating the Nrf2/ARE/HO-1 pathway.