Apigenin Alleviates Allodynia and Hyperalgesia in a Mouse Model of Chemotherapy-Induced Peripheral Neuropathy via Regulating Microglia Activation and Polarization.

BACKGROUND: Apigenin has been reported to exhibit anti-inflammatory and anti-oxidative activities. This study aimed to investigate the protective role of Apigenin on chemotherapy-induced peripheral neuropathy (CIPN). METHODS: CIPN mouse model was established using Paclitaxel treatment. Hot plate and tail prick latency tests were performed to examine the allodynia and hyperalgesia behaviors. Anti-inflammatory and anti-oxidative effects of Apigenin on CIPN were determined by enzyme-linked immunosorbent (ELISA) assay, Western blot, and qRT-PCR. Nuclear recruitment of nuclear factor erythroid 2-related factor 2 (NRF2) was analyzed to evaluate the underlying mechanisms of the protective effects of Apigenin. RESULTS: Apigenin significantly alleviated CIPN-induced nociceptive behaviors of CIPN mice. It also decreased the TNF-α and IL-1ß levels, suppressed oxidative stress and inflammation in the surgical spinal cord tissues. Mechanistically, Apigenin altered the pro-inflammatory and anti-inflammatory phenotypes ratio of microglia through promoting the nuclear recruitment of NRF2 and activating the NRF2/Antioxidant Response Element (ARE) signaling pathway. CONCLUSIONS: In summary, Apigenin relieves CIPN by regulating microglia activation and polarization, which provides a potential therapeutic strategy for CIPN treatment.

Energy intake restriction significantly improves POCD after internal fixation of tibial fractures in mice.

OBJECTIVE: To investigate the effect of energy intake restriction on postoperative cognitive dysfunction (POCD) after internal fixation of tibial fractures in mice. METHODS: Thirty mice were divided into model groups of internal fixation of tibial fractures with 0%, 20%, 30% and 40% energy intake restriction and sham operation group (n = 6). Novel object recognition task and elevated plus maze test were used to assess the ability of recognition memory and anxiety-related behavior before and one week after surgery. The blood samples were collected from mice on days 1, 3 and 7 after surgery, and the mice were euthanized on the 8th day after surgery. RT-PCR and Western blot were employed to detect the expression of AMPK-SIRT1 pathway-related genes and proteins in the hippocampus. ELISA was used to detect the levels of inflammatory factors in the peripheral blood of mice. Hematoxylin-eosin (H&E) staining and immunofluorescence (IF) staining were used to detect the proliferation, differentiation and injury of hippocampal cells. RESULTS: The results showed that 20% and 30% energy intake restriction significantly improved the POCD after internal fixation of tibial fractures in mice. Significantly, 30% energy intake restriction reduced the expression of AP-1, NF-κB, CD45, IBA-1, and inflammatory factors IL-1ß, IL-6, IL-8 and TNF-α, and increased the expression of AMPK and SIRT1 after the operation. H&E and IF staining showed that 30% energy intake restriction reduced postoperative hippocampal neuronal damage. CONCLUSION: Energy intake restriction can significantly improve POCD after internal fixation of tibial fractures in mice and may provide a new treatment paradigm for POCD patients.

Differential effects of remifentanil and sufentanil anesthesia on post-operative pain and cognitive functions.

This study aimed to investigate the differential effects of remifentanil and sufentanil anesthesia on post-operative pain and recovery of cognitive functions following surgical resection of human colon cancer orthotopically transplanted in rats. Human colon cancer cells HT-29 were used to establish a rat model of orthotopically transplanted colon cancer on to the cecal wall of rats. The transplanted tumors were then surgically removed after 5 weeks, using different doses of remifentanil and sufentanil anesthesia. At 24 h after the surgery, von Frey test, hot plate test and voluntary wheel running test were used to evaluated post-operative pain in the rats. Morris water maze test and fear conditioning test were employed to assess cognitive functions. Serum and colon tissues of the rats were also subjected to ELISA to measure levels of stress response factors, while colon tissues were analyzed by RT-PCR and Western blot to measure expression of inflammation response factors and NF-κB pathway-related factors. Sufentanil showed better effect in reducing post-operative pain, while remifentanil showed better recovery of cognitive functions after surgery. In addition, remifentanil resulted in less stress and inflammation response, caused milder activation of NF-κB pathway-related factors after surgery. Remifentanil and sufentanil exhibited differential effects on post-operative pain and recovery of cognitive function. Specifically, remifentanil caused lower stress and inflammation response, associated with dampened activation of the NF-κB pathway. Our results could provide theoretical basis for adopting appropriate analgesic strategy and agents according to the characteristics of individual patients.

The Anti-inflammatory Effect of Dexmedetomidine Administration on Patients Undergoing Intestinal Surgery: A Randomized Study.

BACKGROUND AND OBJECTIVE: Dexmedetomidine is a highly selective α2-adrenergic receptor agonist with sedative, analgesic, anti-sympathetic and stress-reducing effects. It has been widely used as an adjunct for general anesthesia of multiple surgeries. However, the relationship between the utilization of dexmedetomidine in intestinal surgery and the postoperative inflammatory response of patients remains unclear. METHODS: A randomized, controlled, single-blinded clinical trial was performed. Eighty-six patients assigned for intestinal surgery were recruited and were randomly divided into two groups (dexmedetomidine group, n = 40; control group, n = 40) [six participants were excluded due to multiple reasons, such as allergy and drug use history]. The clinical characteristics and physiological outcomes of participants who received different treatments (dexmedetomidine and 0.9% sodium chloride) were collected and analyzed. Blood samples of the two groups were collected before administration (T0), 10 min after pumping dexmedetomidine/saline solution (T1), immediately after the operation started (T2), 30 min after the operation started (T3), and immediately after the operation ended (T4). Enzyme-linked immunosorbent assay (ELISA) was performed to evaluate the proinflammatory factors. RESULTS: Intravenous injection of dexmedetomidine before intestinal surgery decreased a variety of circulating proinflammatory factors. Dexmedetomidine alleviated the stress response and promoted the recovery of cognitive ability among patients undergoing intestinal surgery. CONCLUSION: Dexmedetomidine administration in patients undergoing intestinal surgery inhibited the surgery-induced inflammatory reactions.

Ultrasound-guided stellate ganglion block alleviates stress responses and promotes recovery of gastrointestinal function in patients.

BACKGROUND: This study aimed to investigate the effect of preoperative ultrasound-guided stellate ganglion block (SGB) on the perioperative stress responses and gastrointestinal functions of patients undergoing laparoscopic colorectal cancer surgery. METHODS: A total of 60 colorectal cancer patients were enrolled in study and were randomized to be treated with or without SGB therapy. In the SGB group, patients were injected with 7 mL 0.5% ropivacaine in stellate ganglion under ultrasound guidance before anesthesia. Mean artery pressure (MAP), heart rate (HR), recovery of bowel sound and first exhaust, as well as levels of motilin, gastrin, norepinephrine, cortisol, interleukin-6 (IL-6) and C-reactive protein (CRP) were recorded at various time points. RESULTS: 26 patients in the SGB group and 27 patients in the control group were analyzed. No significant differences in MAP or HR were observed between the two groups before, during and after the surgery. SGB promoted recovery of gastrointestinal functions, as evidenced by earlier recovery of bowel sound and first exhaust, as well as increased motilin and gastrin levels. SGB also attenuated stress responses, as shown in reduced norepinephrine, cortisol, IL-6 and CRP levels. CONCLUSIONS: SGB promotes the recovery of gastrointestinal functions and reduces stress responses of colorectal patients undergoing laparoscopic colorectal cancer surgery.

Effects of Depth of Anesthesia Monitored by IoC on Patients Undergoing Laparoscopic Radical Resection of Colorectal Cancer.

Index of consciousness (IoC) consisting of IoC1 and IoC2, is a new analgesia monitoring indicator in anesthesia evaluation in the laparoscopic radical resection of colorectal cancer. Although the precise anesthetic dosage adjusted by IoC1 has been confirmed to enhance the recovery and reduce the complications of anesthesia, the most appropriate range of IoC2 during anesthesia remains unclear. To investigate the correlation between IoC2 and peri-operative indicators of patients during laparoscopic radical resection of colorectal cancer, the current randomized, controlled, and single-blinded clinical trial was performed. Participants were divided randomly into three groups with different anesthesia depth monitored by IoC2 during their laparoscopic radical resections. Primary outcomes included the dosage of remifentanil. Secondary outcomes included other physiological indexes and complications. The remifentanil dosage and the awakening time increased as IoC2 decreased. The incidences of hypotension and hypoxemia decreased with the elevated IoC2, but the risk of intra-operative awareness also increased. The impact caused by anesthesia to the immune system and health-related life quality of the patients descended with reduced anesthetic level. The IoC2 range of 35-45 could represent the most appropriate anesthetic depth during laparoscopic radical resection, which provides a new perspective for the clinical treatment of colon cancer.

Long noncoding RNA LINC00630 promotes radio-resistance by regulating BEX1 gene methylation in colorectal cancer cells.

This study aimed to analyze the relation between long noncoding RNA (lncRNA) LINCE00630 and radio-resistance and elucidate the underlying mechanism. Relative expression of LINC00630, BEX1, and DNMT3B in colorectal cancer (CRC) cells and clinical samples was determined by real-time PCR. Prognosis in respect of LINC00630 expression was analyzed by Kaplan-Meier survival curve. LINC00630 and BEX1 were specifically silenced by shRNAs. Cell viability and growth were analyzed by MTT and clonogenic assays, respectively. Cell apoptosis was measure by both caspase-3 activity and flow cytometry. Association between EZH2 with LINC00630 and BEX1 promoter was determined by RNA immunoprecipitation and chromatin immunoprecipitation. BEX1 and DNMT3B proteins were quantified by Western blot. We demonstrated the elevated LINC00630 correlated with radio-resistance and poorer prognosis in CRC. Knockdown of LINC00630 significantly improved the sensitivity of CRC cells to irradiation. Mechanistically, LINC00630 in complex with EZH2 negatively regulated BEX1 through promoter DNA methylation. BEX1 silencing greatly restored the cell viability and suppressed cell apoptosis, which were elicited by LINC00630 deficiency in response to irradiation. Our data uncovered the contribution of elevated LINC00630 to radio-resistance in CRC, which was predominately mediated by epigenetically repressed BEX1.

miR-101 down-regulates mTOR expression and attenuates neuropathic pain in chronic constriction injury rat models.

We aimed to demonstrate the effects of microRNA (miR)-101 on neuropathic pain and explore the underlying mechanisms. Rat spinal microglia cells were isolated and inflammatory condition was stimulated by 24-h incubation with lipopolysaccharide (LPS). Rats were divided into 4 groups: sham, chronic constriction injury (CCI), CCI + miR-negative control (miR-NC) and CCI + miR-101 mimics. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) tests were conducted. The mRNA levels of key genes were determined by quantitative real-time polymerase chain reaction. Mammalian target of rapamycin (mTOR) protein level was detected by Western blot. Concentrations of interleukin (IL)-6, IL-1ß and tumor necrosis factor (TNF)-α were examined by ELISA. MiR-101 was downregulated and mTOR was upregulated in lumbar spinal dorsal horns from CCI rats. Targetscan and luciferase reporter assay confirmed that mTOR was direct target of miR101. MiR-101 mimics inhibited LPS-stimulated increase in the levels of IL-6, IL-1ß and TNF-α in primary microglial cells in vitro. In the rat CCI model, miR-101 mimics also suppressed CCI-induced decrease in PWT and PWL and inhibited CCI-induced increase in mRNA and protein levels of IL-6, IL-1ß and TNF-α. In addition, miR-101 downregulated mTOR mRNA and protein expressions in CCI rats. Besides, CCI operation resulted in miR-101 downregulation and mTOR mRNA upregulation in spinal microglia cells in a time-dependent manner. In conclusion, miR-101 had neuropathic pain-attenuating activity through targeting mTOR.

Long noncoding RNA LINC00342 promotes growth of infantile hemangioma by sponging miR-3619-5p from HDGF.

Infantile hemangiomas (IH) are a type of benign vascular neoplasm that may cause permanent scarring. Hemangioma-derived endothelial cells (HemECs) are commonly used as an in vitro model to study IH. Long noncoding RNA is a type of RNA transcript longer than 200 nucleotides that does not encode any protein. LINC00342 was discovered to regulate proliferation and apoptosis in nonsmall cell lung cancer. However, the role of LINC00342 in IH has never been reported before. Expressions of LINC00342 and miR-3619-5p were detected in proliferating versus normal skin tissues. Colony formation and Cell-Couting Kit 8 assays were carried out to study the effects on cell proliferation after knockdown and overexpression of LINC00342, respectively. Meanwhile caspase-3 activity and nucleosomal fragmentation assay were applied to detect cell apoptosis. Micro-RNA binding sites on LINC00342 and hepatoma-derived growth factor (HDGF) were predicted and confirmed via dual-luciferase reporter assay. Biotin RNA pulldown assay was used to verify the direct binding between RNA molecules. LINC00342 enhanced proliferation and inhibited apoptosis in HemECs. MiR-3619-5p targeted both LINC00342 and HDGF, where LINC00342 sponged miR-3619-5p and positively regulated HDGF. HDGF knockdown rescued the effects of LINC00342 on HemECs. The LINC00342-miR-3619-5p-HDGF signaling pathway could regulate cell proliferation and apoptosis in HemECs.NEW & NOTEWORTHY The role of LINC00342 in infantile hemangiomas has not yet been elucidated. This paper highlights the regulatory role of LINC00342 in cell proliferation and apoptosis in hemangioma-derived endothelial cells and the underlying molecular mechanisms. The findings would provide potential target for treatment of infantile hemangiomas.

Administration of Curcumin Alleviates Neuropathic Pain in a Rat Model of Brachial Plexus Avulsion.

BACKGROUND/AIMS: Brachial plexus avulsion (BPA) generally causes a chronic persistent pain that lacks efficacious treatment. Curcumin has been found to possess anti-inflammatory abilities. However, little is known about the mechanisms and effects of curcumin in an animal model of BPA. METHODS: Mechanical withdrawal thresholds (MWT) were examined by von Frey filaments. Cold allodynia was tested by the acetone spray test. The levels of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in rat spinal cords were analyzed by the enzyme-linked immunosorbent assay, and the expression levels of c-Fos and nerve growth factor (NGF) were measured by Western blot. The expression level of glial fibrillary acidic protein (GFAP) was observed by immunofluorescence and Western blot. RESULTS: After curcumin treatment, the MWT showed a significant increase when compared to the BPA group on both hind paws. A remarkable decrease of paw-withdrawal response frequency was observed compared with the BPA group. In addition, curcumin treatment significantly decreased the levels of TNF-α and IL-6 in rat spinal cords that were exceedingly upregulated in the BPA group. The protein levels of c-Fos and NGF were decreased by treatment with curcumin compared with the corresponding protein levels in the BPA group. Besides, curcumin reduced the number of GFAP positive cells and GFAP expression. CONCLUSIONS: Our findings suggest that curcumin significantly extenuates the BPA-induced pain and inflammation by reducing the expression level of proinflammatory cytokines and pain-associated proteins and inhibiting the activity of astrocytes.