[Improvement effect of cinnamaldehyde on reserpine-induced Parkinson's disease rat model].

In order to study the neuroprotective mechanism of cinnamaldehyde on reserpine-induced Parkinson's disease(PD) rat models, 72 male Wistar rats were randomly divided into blank group, model group, Madopar group, and cinnamaldehyde high-, medium-, and low-dose groups. Except for the blank group, the other groups were intraperitoneally injected with reserpine of 0.1 mg·kg~(-1) once every other morning, and cinnamaldehyde and Madopar solutions were gavaged every afternoon. Open field test, rotarod test, and oral chewing movement evaluation were carried out in the experiment. The brain was taken and fixed. The positive expression of dopamine receptor D1(DRD1) was detected by TSA, and the changes in neurotransmitters such as dopamine(DA) and 3,4-dihydroxyphenylacetic acid(DOPAC) in the brain were detected by enzyme-linked immunosorbent assay(ELISA). The protein and mRNA expression levels of tyrosine hydroxylase(TH) and α-synuclein(α-Syn) in substantia nigra(SN) were detected by RT-PCR and Western blot. The results showed that after the injection of reserpine, the hair color of the model group became yellow and dirty; the arrest behavior was weakened, and the body weight was reduced. The spontaneous movement and exploration behavior were reduced, and the coordination exercise ability was decreased. The number of oral chewing was increased, but the cognitive ability was decreased, and the proportion of DRD1 positive expression area in SN was decreased. The expression of TH protein and mRNA was down-regulated, and that of α-Syn protein and mRNA was up-regulated. After cinnamaldehyde intervention, it had an obvious curative effect on PD model animals. The spontaneous movement behavior, the time of staying in the rod, the time of movement, the distance of movement, and the number of standing times increased, and the number of oral chewing decreased. The proportion of DRD1 positive expression area in SN increased, and the protein and mRNA expression levels of α-Syn were down-regulated. The protein and mRNA expression levels of TH were up-regulated. In addition, the levels of DA, DOPAC, and homovanillic acid(HVA) neurotransmitters in the brain were up-regulated. This study can provide a new experimental basis for clinical treatment and prevention of PD.

Ginseng promotes the function of intestinal stem cells through the Wnt/ß-catenin signaling pathway in D-galactose-induced aging mice.

BACKGROUND: Intestinal stem cells (ISCs) are the reservoir source of various types of intestinal cells, and the decline of stem cell function in the gut may be a potential factor for aging-related disease. The present study aimed to explore the regulatory mechanisms of Panax ginseng C.A.Meyer (Araliaceae, Panax genus) that could restore gut aging by enhancing intestinal function and regulating ISCs in aging mice based on the Wnt/ß-catenin signaling pathway. METHODS: A total of 60 ICR male mice were randomly divided into control, model, metformin, and ginseng water decoction (GWD) 3.6, 1.8, and 0.9 g/kg groups. The aging model was induced by 1 % D-galactose (s.c. 0.1 mL/10 g) for 28 days. Moreover, GWD was given to aging mice intragastrically (i.g.) once a day for 28 successive days. The learning memory ability, pathological status, and function in the ileum tissue, the activity of digestive enzymes, and short-chain fatty acid (SCFA) content in the colon were evaluated, and the related mechanism was investigated. RESULTS: Ginseng can decrease the escape latency time and increase the swimming speed and the number of crossing platforms in aging mice. Moreover, the pathology of ileum tissue improved, the length of the intestinal villi increased, and the width of the villi and the depth of the crypts decreased. The activities of trypsin, α-amylase, and lipase increased in duodenal content and intestinal mucosa. In the colon, the content of SCFA, such as acetic acid, propionic acid and butyric acid, increased, indicating that ginseng significantly improves intestinal function impairment. The mRNA expressions and protein levels of ß-catenin, C-myc, GSK-3ß, Lgr5, and Olfm4 were upregulated in the ginseng group. CONCLUSIONS: Ginseng improves intestinal function and regulates the function of ISCs in order to protect intestinal health by activating the Wnt/ß-catenin signaling pathway in aging mice.

Prophylactic Effects of Hemp Seed Oil on Perimenopausal Depression: A Role of HPA Axis.

Hemp seed, the dried fruit of Cannabis sativa L. (Moraceae), has been extensively documented as a folk source of food due to its nutritional and functional value. This study evaluated the antidepressant effect of hemp seed oil (HSO) during its estrogen-like effect in Perimenopausal depression (PMD) rats induced by ovariectomy combined with chronic unpredictable mild stress (OVX-CUMS). Female SD rats (SPF, 10 weeks, sham operated group, ovariectomy (OVX) model group, ovariectomy - chronic unpredictable mild stress (OVX-CUMS) group, HSO + OVX-CUMS group, fluoxetine (FLU) + OVX-CUMS group, n=8) were subjected to treatment with HSO (4.32 g/kg) or fluoxetine (10 mg/kg) for 28 days (20 mL/kg by ig). Sucrose preference test (SPT), forced swimming test (FST), open field test (OFT), estrogen receptor α (ERα) and estrogen receptor ß (ERß) expression, estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), cortisol (CORT), adrenocorticotropic hormone (ACTH), corticotropin releasing hormone (CRH), norepinephrine (NE), 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5HIAA) levels are measured to evaluate the function of the hypothalamic-pituitary-ovarian (HPO) and hypothalamic-pituitary-adrenal (HPA) axis. The results showed that OVX-CUMS significantly decrease sucrose preference rate in SPT, increase immobility time in FST and OFT, and decrease movement distance and stand-up times in OFT. HSO treatment significantly improves depression-like behaviors, upregulates the expression of ERα and ERß, improves HPO axis function by increasing E2 levels and decreasing FSH and LH levels, reverses HPA axis hyperactivation by decreasing CORT, ACTH, and CRH levels, and upregulates NE, 5-HT, and 5HIAA levels in model rats. The findings suggested that HSO could improve depression-like behavior in OVX-CUMS rats by regulating HPO/HPA axis function and neurotransmitter disturbance.

Neuroprotective effect of 20 (S) - Protopanaxadiol (PPD) attenuates NLRP3 inflammasome-mediated microglial pyroptosis in vascular dementia rats.

20(S)-protopanaxadiol (PPD), one of the ginsenosides from Panax ginseng, has been reported to improve performance with dementia. This study aimed to investigate the neuroprotective effect of PPD attenuating NLRP3 inflammasome-mediated microglial pyroptosis in vascular dementia (VD) rats induced by bilateral common carotid artery ligation (2-VO). Male Sprague-Dawley rats (SPF, 150-180 g, n = 10/group) were randomly divided into PPD (20, 10, 5 mg/kg, subcutaneous injection once per day for 3 weeks), model, and vehicle-sham group. It was found that PPD significantly reversed 2-VO-induced cognitive impairment by decreasing escape latency and spontaneous alternation and increasing the number of crossing platforms, showing memory-improving effects. PPD improved the pathological morphology of brain tissue in VD rats. PPD significantly reduced the cerebral infarction area and the activation of microglia in the cortex and hippocampal DG, CA1, and CA3 area. Moreover, PPD could attenuate NLRP3 inflammasome-mediated microglial pyroptosis, inhibit the positive expression of NLRP3, decrease IL-1ß, and IL-18 levels, and increase IL-10 levels in the brain cortex. PPD also significantly alleviated the neurotoxicity by decreasing the Aß and p-Tau in hippocampal DG, CA1, and CA3 areas. In addition, the levels of NLRP3, ASC, and IL-1ß in the cortex, APP, BACE1, and p-Tau in the hippocampus were significantly reduced by PPD. These results suggested that PPD hinders microglial activation to alleviate neuroinflammation of NLRP3 inflammasome and inhibits neurotoxicity of Aß deposition and Tau phosphorylation in 2-VO-induced VD rats.

[Neuroprotective effect of cinnamaldehyde in MPTP-induced mouse model of subacute Parkinson's disease].

This paper aims to explore the neuroprotective effect of cinnamaldehyde(CA) in mice with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced subacute Parkinson's disease(PD) and the mechanism. To be specific, male C57 BL/6 mice(n=72, SPF) were randomized into control group, model group, positive control(madopar 0.1 mg·g~(-1)) group, and low-dose, me-dium-dose, and high-dose CA groups(0.15, 0.30, 0.60 mg·g~(-1)). MPTP(intraperitoneal injection, 0.03 mg·g~(-1), once a day for 5 days) was used to induce subacute PD in mice except for the control group. The administration began from the day of modeling and lasted 19 days. On the 0 th, 12 th, and 19 th day, the open field test, pole test, and rotarod test were carried out. After the tests, the mice were killed and brains were separated. In addition, the organ index was measured. The number of cells in substantia nigra(SN) in the midbrain of MPTP-induced PD model mice was detected based on hematoxylin and eosin(HE) staining. The levels of tyrosine hydroxylase(TH)-and α-synuclein(α-Syn)-positive cells in SN were determined by immunohistochemical staining, and the protein levels of TH and α-Syn in SN by Western blot. The results showed that the MPTP-stimulated mice had abnormal behaviors such as erect hair, arched back, rigidity of the tail, slow movement, and tremor, decreased number of crossings and rearing, increased frequency of urination and defecation, longer time of pole climbing, and shorter time of staying on the rotating rod. In addition, the mice showed obvious damage of neurons in the SN and reduced neuron cells in irregular arrangement with some shrinking. In addition, the average optical density of TH in SN decreased and that of α-Syn increased. All these suggested the successful modeling. CA displayed obvious therapeutic effect on the PD mice, as manifested by the increased number of crossings and rearing, decreased frequency of urination and defecation, shorter time of climbing pole, longer time of staying on the rotating rod, and more neuron cells in the SN with a few pykno-tic cells. Moreover, CA significantly alleviated the decrease of TH and the overexpression of α-Syn in SN. As a result, the MPTP-induced injury of dopaminergic neurons was alleviated. The performance of 0.3 mg·g~(-1) CA was the best. This study is expected to lay a scientific basis for the development of CA products.

Panax ginseng root, not leaf, can enhance thermogenic capacity and mitochondrial function in mice.

Context: Panax ginseng C. A. Meyer (Araliaceae) root and leaf have always been considered in the traditional theory as hot and cold properties, respectively.Objective: To clarify the hot and cold properties of ginseng root and leaf from a thermodynamic viewpoint.Materials and methods: Thirty ICR male mice were randomly assigned to control (water), ginseng root group (GRP) and ginseng leaf group (GLP) with a concentration of 0.075 g/mL; the volume was 0.1 mL/10 g (body mass) per day by intragastric administration for 20 days. Ultra-Performance Liquid Chromatography (UPLC) was used to determine quality control through seven ginsenosides contained in ginseng root and leaf. Rest metabolic rate (RMR) and energy expenditure were monitored every 9 days by TSE System. At the 20th day, serum T3 or T4, liver or brown adipose tissue (BAT) mitochondrial respiration were investigated.Results: The quality control of GRP and GLP were within requirements of 2015 China Pharmacopoeia. The RMR (mLO2/h) in GLP (47.95 ± 4.20) was significantly lower than control (52.10 ± 4.79) and GRP (55.35 ± 4.48). Mitochondrial protein concentration and respiration were significantly increased in GRP (BAT, 79.12 ± 2 .08 mg/g, 239.89 ± 10.24 nmol O2/min/g tissue; Liver, 201.02 ± 10.89, 202.44 ± 3.24) and decreased in GLP (BAT, 53.42 ± 3.48, 153.49 ± 5.58; Liver, 138.69 ± 5.69, 104.50 ± 6.25) compared with control.Conclusions: The hot and cold properties of ginseng root and leaf are correlated with thermogenic capacity and mitochondrial function of BAT and liver, which deserve to further research.

Metabolite identification of gut microflora-cassia seed interactions using UPLC-QTOF/MS.

Cassia seed is the dried ripe seed of Cassia obtusifolia L. or Cassia tora L., which is widely used as a food or traditional Chinese medicine. The aim of the present study was to detect the components and metabolites in the culture of human or rat intestinal microflora suspension with the water decoction of cassia seed in vitro, using an ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry system equipped with a negative ion scan mode. Initially, ellagic acid was identified in the cassia seed decoction. Subsequently, six different metabolites, including urolithin (uro)-A, uro-B, uro-D, uro-M6, uro-M7 and uro-B-glucuronide (glur), were detected after co-culture of the cassia seed decoction with intestinal microflora, but not in the cassia seed decoction alone. Uro-M6, uro-M7, uro-A and uro-B were common metabolites in the culture of human or rat intestinal microflora suspension with the water decoction of cassia seed. However, uro-D was only detected in the culture of rat intestinal microflora suspension with the water decoction of cassia seed, and uro-B-glur was only detected in the culture of human intestinal microflora with the water decoction of cassia seed. The uro and intermediate metabolites were produced by ellagic acid in the cassia seed decoction under the action of the intestinal microflora. The production of metabolites might be related to the abundance and diversity of the intestinal microflora in humans and rats. The present study provided rationale for further pharmacological and clinical studies on the mechanisms of action of cassia seeds.

Elevation of synaptic protein is associated with the antidepressant-like effects of ferulic acid in a chronic model of depression.

Ferulic acid is a hydroxycinnamic acid that widely presents in plant cell wall components. It has been demonstrated that ferulic acid can reverse depressive-like behaviors in both forced swimming test and tail suspension test. However, it is unclear whether chronic ferulic acid treatment can ameliorate the depressive-like behaviors in chronic unpredictable mild stress (CUMS). Because of the putative relationship between neurotrophic system and antidepressant-like activity, we also investigated the effects of chronic ferulic acid on the brain-derived neurotrophic factor (BDNF), postsynaptic protein PSD95, presynaptic protein synapsin I in both prefrontal cortex and hippocampus. The results showed that ferulic acid significantly alleviated CUMS-induced depressive-like behaviors in sucrose preference test and forced swimming test. In addition, ferulic acid significantly up-regulated the levels of BDNF, PSD95 and synapsin I in the prefrontal cortex and hippocampus. The present data indicated that ferulic acid exerted the antidepressant-like effects on behaviors by increasing neurotrophin-related synaptic protein levels in CUMS mice.

Limits to sustained energy intake. XI. A test of the heat dissipation limitation hypothesis in lactating Brandt's voles (Lasiopodomys brandtii).

The maximum rate of sustained energy intake (SusEI) may limit reproductive effort, thermoregulatory capability and other aspects of an animal's energy expenditure. Consequently, factors that limit SusEI are of interest. The ;heat dissipation limitation hypothesis' suggests that maximum SusEI during lactation is limited by the capacity to dissipate body heat generated as a by-product of processing food and producing milk. In the present study, we tested the heat dissipation limitation hypothesis in lactating Brandt's voles (Lasiopodomys brandtii). Female voles were mated and pregnant at 21(+/-1) degrees C. A random sample of animals was transferred into a hot room 30(+/-1) degrees C on the day of parturition. The energy intake of lactating voles at 30 degrees C was always lower than that at 21 degrees C. At peak lactation food intake was 3.3 g day(-1) lower at 30 degrees C than at 21 degrees C. There was no significant difference in digestibility. With similar mean litter sizes (7.26+/-0.46 pups at 21 degrees C and 7.78+/-0.39 pups at 30 degrees C at the beginning of parturition, 6.83+/-0.51 pups at 21 degrees C and 7.73+/-0.50 pups at 30 degrees C at weaning), the milk energy output of mothers, evaluated from the difference between metabolizable energy intake and daily energy expenditure measured by doubly labelled water, at 30 degrees C was 23.3 kJ day(-1) lower than that at 21 degrees C on days 14-16 of lactation. As for reproductive performance, there was a difference in the response to the higher temperature between mothers raising large and those raising small litters. For small litters (<7) there was no significant change in litter mass, but for large litters (

[Expression of ETV5 and MMP-7 in early stage cervical squamous cell carcinoma and its role in lymphatic metastasis].

BACKGROUND & OBJECTIVE: Transcription factor Ets variant gene 5 (ETV5) participates in tumor metastasis, and matrix metalloproteinase-7 (MMP-7) also plays an important role in tumor invasion and metastasis. This study was to investigate the expression of ETV5 and MMP-7 in cervical squamous cell carcinoma, and explore the roles of ETV5 and MMP-7 in invasion and metastasis of early stage cervical squamous cell carcinoma. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of ETV5 and MMP-7 mRNA in 86 samples of cervical squamous cell carcinoma and 21 samples of normal cervix, and immunohistochemistry was used to detect the expression of MMP-7 protein in these samples. RESULTS: The mRNA levels of ETV5 and MMP-7 were significantly higher in cervical cancer tissues than in normal cervix tissues (0.507+/-0.073 vs. 0.201+/-0.057, P<0.01; 0.488+/-0.078 vs. 0.305+/-0.064, P<0.01), and were significantly higher in nodal metastasis group than in no nodal metastasis group (0.538+/-0.063 vs. 0.496+/-0.073, P<0.05; 0.542+/-0.066 vs. 0.468+/-0.072, P<0.01). The protein level of MMP-7 was significantly higher in cervical cancer tissues than in normal cervix tissues (3.826+/-1.427 vs. 1.714+/-0.383, P<0.01), and was significantly higher in nodal metastasis group than in no nodal metastasis group (4.935+/-1.614 vs. 2.623+/-1.086, P<0.01). The positive rate of MMP-7 protein was significantly higher in lymph node metastasis group than in no lymph node metastasis group (65.21% vs. 36.51%, P<0.01). ETV5 expression was positively correlated to MMP-7 expression (r=0.221, P<0.05). CONCLUSION: ETV5 and MMP-7 are highly expressed in metastatic cervical squamous cell carcinoma, and may be served as markers in estimating the invasive and metastatic potential of cervical squamous cell carcinoma.

[Effect of compound recipe Gengniankang on senile sexual hormone and expression of estrogen receptor in bone of climacteric female rats].

OBJECTIVE: To compare the therapeutic effect of Compound Recipe Gengniankang ( GNK) with that of hormone replacement treatment (HRT) on climacteric female rats with osteoporosis, and to investigate the roles of estrogen and estrogen receptors in the mechanism of osteoporosis. METHODS: Climacteric female rats with osteoporosis were chosen and divided into three groups (GNK group, HRT group and control group). Apoptosis of ovarian granulose cells was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay. Serum level of estradiol (E(2)), follicle stimulating hormone (FSH), luteinizing hormone (LH) were determined by the method of radioimmunoassay (RIA). Reverse transcriptase polymerase chain reaction (RT-PCT) technology was used to evaluate the expression of estrogen receptor (ER) in bone. Bone mineral density (BMD) was measured by double energy X-ray absorption (DEXA). RESULTS: In the climacteric rats, BMD, serum E(2), ER mRNA expression in bone decreased remarkably, and serum FSH, LH and apoptosis of ovarian granulose cells increased obviously. After treating with GNK, all the indexes were reversed except serum E(2). The increase of E(2) was not significant. CONCLUSION: GNK is effective on climacteric osteoporosis female rats. Its role is performed not by increasing serum E(2) but by enhancing ER in the bone and inhibiting apoptosis of ovarian granulose cells. GNK can deter further exhaustion of ovarian function.

[Experimental study on regulatory effect of compound gengniankang on endocrine and immune functions in aged female rats with osteoporosis].

OBJECTIVE: To observe the effect of compound gengniankang (GNK) in regulating the endocrine and immune functions in aged female rats. METHODS: Aged female rats with osteoporosis were selected as the object for observation and healthy young rats were taken for control. Animals were administered by GNK and nilestriol respectively, through gastric perfusion, for 3 months to observe the therapeutic effect of drug treatment on osteoporosis and the regulatory effect on endocrine and immune function. Bone mineral density (BMD) was measured by double energy X-ray absorption technique, serum levels of estradiol (E2), follicule-stimulating hormone (FSH) and luteinizing hormone (LH) were determined by RIA, T-cell subsets and apoptosis in spleen were detected by flow cytometry. RESULTS: In aged rats with osteoporosis, the BMD decreased, serum level of E2 lowered, FSH and LH levels raised, splenic CD4+, CD4+/CD8+ significantly decreased and T-cell apoptosis rate significantly elevated. GNK could increase the BMD, lower the FSH and LH levels, but showed no significant effect on E2 level. It could increase the CD4+ and CD4+/CD8+ ratio to nearby the normal range, and reduce the apoptosis of T-cells. CONCLUSION: GNK has therapeutic effect on osteoporosis in aged rats, and is able to regulate the endocrine and enhance the immune function in organism.