Susu Lin1, Qiaoqiao Li1, Shanshan Jiang1, Zijin Xu1, Yu Jiang1, Ling Liu1, Jinyan Jiang1, Yingpeng Tong2, Ping Wang3. 1. College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. 2. Institute of Natural Medicine and Health Products, School of Advanced Study, Taizhou University, Taizhou, 318000, People's Republic of China. 3. College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. Electronic address: wangping45@aliyun.com.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: This study aimed at determining the effects of saffron on depression as well as its neuroprotective and pharmacological effects on the intestinal function of crocetin in mice exposed to chronic restraint stress. MATERIALS AND METHODS: Chronic stress was induced in two-week-old ICR mice by immobilizing them for 6 h per day for 28 days. The mice were orally administered with crocetin (20, 40, 80 mg/kg), fluoxetine (20 mg/kg) or distilled water. The treatments were administered daily and open field and tail suspension tests were performed. Immunofluorescent and Western-bolt (WB) assays were conducted to determine the expression of mitogen-activated protein kinase phosphatase-1 (MKP-1), the precursor of brain-derived neurotrophic factor (proBDNF), extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated cAMP response element-binding (CREB) protein in the hippocampus. Serum levels of dopamine (DA), proBDNF, MKP-1 and CREB were measured by Elisa kits. High-throughput sequencing was carried out to analyze the composition of intestinal microbiota. RESULTS: Crocetin ameliorated depressive-like behaviors caused by chronic restraint stress-induced depressive mice. It significantly attenuated the elevated levels of MKP-1, proBDNF, alanine transaminase, aspartate transaminase and increased the serum levels of DA as well as CREB. Histopathological analysis showed that crocetin suppressed hippocampus injury in restraint stress mice by protecting neuronal cells. Immunofluorescent and WB analysis showed elevated expression levels of ERK1/2, CREB and inhibited expression levels of MKP-1, proBDNF in the hippocampus. The intestinal ecosystem of the crocetin group partially recovered and was close to the control group. CONCLUSIONS: Crocetin has neuroprotective properties and ameliorates the effects of stress-associated brain damage by regulating the MKP-1-ERK1/2-CREB signaling and intestinal ecosystem.
ETHNOPHARMACOLOGICAL RELEVANCE: This study aimed at determining the effects of saffron on depression as well as its neuroprotective and pharmacological effects on the intestinal function of crocetin in mice exposed to chronic restraint stress. MATERIALS AND METHODS: Chronic stress was induced in two-week-old ICR mice by immobilizing them for 6 h per day for 28 days. The mice were orally administered with crocetin (20, 40, 80 mg/kg), fluoxetine (20 mg/kg) or distilled water. The treatments were administered daily and open field and tail suspension tests were performed. Immunofluorescent and Western-bolt (WB) assays were conducted to determine the expression of mitogen-activated protein kinase phosphatase-1 (MKP-1), the precursor of brain-derived neurotrophic factor (proBDNF), extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated cAMP response element-binding (CREB) protein in the hippocampus. Serum levels of dopamine (DA), proBDNF, MKP-1 and CREB were measured by Elisa kits. High-throughput sequencing was carried out to analyze the composition of intestinal microbiota. RESULTS:Crocetin ameliorated depressive-like behaviors caused by chronic restraint stress-induced depressivemice. It significantly attenuated the elevated levels of MKP-1, proBDNF, alanine transaminase, aspartate transaminase and increased the serum levels of DA as well as CREB. Histopathological analysis showed that crocetin suppressed hippocampus injury in restraint stressmice by protecting neuronal cells. Immunofluorescent and WB analysis showed elevated expression levels of ERK1/2, CREB and inhibited expression levels of MKP-1, proBDNF in the hippocampus. The intestinal ecosystem of the crocetin group partially recovered and was close to the control group. CONCLUSIONS:Crocetin has neuroprotective properties and ameliorates the effects of stress-associated brain damage by regulating the MKP-1-ERK1/2-CREB signaling and intestinal ecosystem.