The role of Src-family kinases in long-term potentiation in rat spinal dorsal horn

AIM: To investigate the role of Src-family kinases (SFKs) in long-term potentiation (LTP) of C-fiber-evoked potentials in rat spinal dorsal horn and the underlying mechanism. METHODS: The effect of SFKs inhibitors on spinal LTP induced by high-frequency stimulation of sciatic nerve was determined by electrophysiological method. The levels of phosphorylated SFKs (p-SFKs) in spinal dorsal horn were measured by Western blotting at different time points after LTP induction. The cell types that expressed p-SFKs following LTP induction were determined by double-labeled immunofluorescence staining. RESULTS: Electrophysiological data revealed that pretreatment with SFKs inhibitors (PP2 or SU6656) enabled high-frequency stimulation to induce long-term depression (LTD) rather than LTP in spinal dorsal horn. Western blotting analysis showed that the level of p-SFKs in ipsilateral spinal dorsal horn increased at 15 min after LTP induction. Double-labeled immunofluorescence staining demonstrated that p-SFKs were highly restricted to spinal microglia. CONCLUSION: SFKs in microglia play a critical role in the induction of LTP in spinal dorsal horn. Inhibition of SFKs and the downstream molecules may be helpful for curing neuropathic pain.     

Role of ERKⅠ/Ⅱ in the induction and maintenance of the spinal LTP

AIM: The role of external-signal regulated kinaseⅠ/Ⅱ(ERKⅠ/Ⅱ) in the induction and maintenance of spinal long-term potentiation (LTP) is evaluated.METHODS: The C-fiber evoked field potentials were recorded at the superficial layers of spinal dorsal horn at the lumbar enlargement.RESULTS: (1) PD98059 (100 μmol/L) or SL327 (200 μmol/L), a selective MEK inhibitor, completely blocked LTP induction when applied at 30 min prior to tetanic stimulation.(2) PD98059 or SL327 reversed spinal LTP in a time-dependent manner.At 15 min after LTP induction, PD98059 (100 μmol/L) or SL327 (200 μmol/L) reversed LTP completely, and at 30 min after LTP induction, the inhibitory rate of spinal LTP inhibited by PD98059 or SL327 was 62.5% and 75.0%， respectively.At 1 h after LTP induction, however, the same concentration of PD98059 or SL327 did not affect the spinal LTP.CONCLUSION: Activation of ERKⅠ/Ⅱ in spinal dorsal horn may be crucial for the induction and the early-phase maintenance of LTP of C-fiber evoked field potentials.     

Protein kinase C is involved in the induction and early-phase maintenance of long-term potentiation of C-fiber evoked field potentials in spinal dorsal horn of adult rats

AIM: The role of protein kinase C (PKC) in the induction and maintenance of spinal long-term potentiation (LTP) was evaluated. METHODS: The C-fiber evoked field potentials were recorded at the superficial layers of spinal dorsal horn at the lumbar enlargement. RESULTS: (i) Chelerythrine （200 μmol/L） or G 6983 (100 μmol/L), a selective PKC inhibitor, completely blocked LTP induction. (ii) Chelerythrine or G 6983 reversed spinal LTP in a time-dependent manner. 15 min after LTP induction, chelerythrine （200 μmol/L） and G 6983 (100 μmol/L) depressed LTP to baseline in all tested rats. The same concentration of chelerythrine and G 6983, applied at 3 h after LTP induction, did not affect LTP. CONCLUSION: PKC in spinal dorsal horn may be crucial for the induction and the early-phase maintenance of LTP of C-fiber evoked field potentials.     

PKA is required in the induction and early-phase maintenance of LTP of C-fiber evoked field potentials in spinal dorsal horn of adult rats

AIM: The role of cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) in the induction and maintenance of spinal long-term potentiation (LTP) was evaluated. METHODS: The C-fiber evoked field potentials were recorded at the superficial layers of spinal dorsal horn at the lumbar enlargement. RESULTS: (1) 8-Br-cAMP induced LTP of C-fiber evoked field potentials and 8-Br-cAMP-induced LTP occludes tetanus-induced LTP. (2) Rp-CPT-cAMPS, an inhibitor of PKA, blocked the induction of spinal LTP and reversed established LTP of C-fiber evoked field potentials in a time-dependent manner. (3) In the presence of anisomycin, a protein synthesis inhibitor, the potentiation induced by 8-Br-cAMP was completely blocked. (4) PD98059, a selective inhibitor of mitogen-activated protein kinase (MAPK),completely blocked the 8-Br-cAMP-induced LTP.CONCLUSION: Activation of PKA signal pathway in spinal dorsal horn may be crucial for the induction and the early-phase maintenance of LTP of C-fiber evoked field potentials.     

Role of interleukin-1β in spinal LTP in rats with neuropathic pain

AIM: To investigate the role of interleukin-1β (IL-1β) in the long-term potentiation (LTP) of C-fiber-evoked field potentials in rats with neuropathic pain. METHODS: The rat model of neuropathic pain was produced by spared nerve injury (SNI) of sciatic nerve or the method of lumbar 5 ventral root transection (L5 VRT). The effect of exogenous IL-1β on C-fiber-evoked field potentials of spinal dorsal horn was tested in both intact rats and the rats with neuropathic pain. The roles of p38 MAPK and NF-κB in the process were also evaluated. RESULTS: IL-1β at concentration of 500 μg/L affected neither basal synaptic transmission mediated by C-fiber nor spinal LTP induced by high frequency stimulation in intact rats. However, low concentration (5 μg/L) of IL-1β induced LTP of C-fiber-evoked field potentials in the rats with neuropathic pain. Pretreatment with either p38 MAPK inhibitor (SB203580) or NF-κB inhibitor (PDTC) completely blocked LTP induced by IL-1β. CONCLUSION: Exogeneous IL-1β might induce spinal LTP of C-fiber-evoked field potentials in the rats with neuropathic pain. p38 MAPK and NF-κB may be involved in the process.     

Effects of P2X4 receptor in spinal microglia on rrTNF-induced pathological pain

AIM: To investigate the effects of P2X₄ receptor on peri-sciatic administration of recombinant rat TNF-α (rrTNF)-induced mechanical allodynia. METHODS: Male Sprague-Dawley rats (180~200 g) were used in the experiments. The levels of P2X₄ receptor on day 3, day 7 and day 14 after peri-sciatic administration of rrTNF were examined by Western blot, and the location of P2X₄ receptor in the spinal dorsal horn was observed by double immunofluorescence staining. The changes of 50% paw-withdrawal thresholds of the rat were detected by behavioral test, and the level of TNF-α in the spinal dorsal horn was also examined by Western blot when TNP-ATP was intrathecally injected before the administration of rrTNF. RESULTS: Compared with control group, the expression of P2X₄ receptor in the spinal dorsal horn on the ipsilateral side significantly increased on day 3, day 7 and day 14 (P<0.01) after rrTNF (100 ng/L) administration. P2X₄ receptor was co-localized only with microglia, but not with neurons or astrocytes. Intrathecal injection of TNP-ATP before rrTNF administration prevented mechanical allodynia induced by rrTNF and inhibited the upregulation of TNF-α in the spinal dorsal horn. CONCLUSION: P2X₄ receptors in microglia may be involved in rrTNF-induced mechanical allodynia by the upregulation of TNF-α in the spinal dorsal horn.     

Effects of mibefradil administered intrathecally on the expression of nNOS in spinal cord of the rats following chronic compression of dorsal root ganglion

AIM: To investigate the effect of mibefradil administered intrathecally on the pain threshold and the expression of neuron nitric oxide synthase (nNOS) in the spinal cord of the rats following chronic compression of dorsal root ganglion. METHODS: The chronic compression of dorsal root ganglion (CCD) model was adopted. Three different doses of mibefradil, a T-type calcium channel blocker, were administered singly intrathecally to the rats after CCD 5 days. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were determined with von Frey hair and thermal radiation apparatus before mibefradil administered and after mibefradil administered 30 min, 60 min, 120 min, 240 min, and 480 min. Changes of nNOS expression in neurons of the spinal dorsal horn were detected by immunohistochemistry method, and the number of the nNOS positive neurons was counted under microscope. RESULTS: MWT and TWL declined significantly from the first day to the 21st day after CCD. Mibefradil administered intrathecally attenuated significantly both mechanical and thermal hyperagesia of the rats, and markedly inhibited the increase in the nNOS expression. CONCLUSION: These data show that T type calcium channel plays a key role in the CCD-induced mechanical and thermal hyperalgesia and possibly involves in the nNOS expression in the spinal.     

Effect of curcumin on p-ERK1/2-AP-1 cascade and diabetic neuropathic pain in rats

AIM: To evaluate the role of p-ERK1/2-AP-1 cascade in the process of curcumin against diabetic neuropathic pain (DNP) in rats.METHODS: Ninety-six male Sprague-Dawley rats were randomly divided into 4 groups (n=24): normal control group, DNP group, DNP with solvent group and DNP with curcumin (100 mg/kg) group. The rat model of diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ, 75 mg/kg). Mechanical allodynia and thermal hyperalgesia were tested by mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) 2 weeks after induction,respectively. The diabetic rats were treated with curcumin (100 mg·kg^-1·d^-1, ip) for 2 weeks. The conditions of hyperalgesia and allodynia were determined 2 d before STZ injection, 14 d after STZ injection, and 3 d, 7 d, 14 d after administered with curcumin. The change of p-ERK1/2 was measured by the methods of Western blotting and immunohistochemistry. The expression of AP-1 in spinal cord dorsal horn and dorsal root ganglion (DRG) was detected by electromobility shift assay (EMSA).RESULTS: Compared with normal control group, the rats in DNP group developed hyperglycemia and a decrease in MWT and TWL associated with an increase in the activity of p-ERK1/2 and AP-1 in dorsal horn and DRG(P<0.05). Compared with DNP group, 7-day treatment with curcumin significantly attenuated mechanical allodynia and thermal hyperalgesia, and these effects were correlated with inhibiting the hyper-activation of p-ERK1/2 and AP-1 14 days after treatment with curcumin (P<0.05).CONCLUSION: Curcumin has beneficial effects on hyperalgesia in STZ-induced peripheral neuropathic pain. Activation of p-ERK1/2 and AP-1 may be the key mechanism of DNP in spinal cord and DRG.     

Roles of Akt pathway and cell apoptosis in leptin-mediated chronic morphine antinociceptive tolerance

AIM: To explore the roles of Akt (also called protein kinase B) and active caspase-3 in the leptin-mediated chronic morphine antinociceptive tolerance in rats. METHODS: A model of chronic morphine antinociceptive tolerance was established in the SD rats. The protein levels of spinal Akt and cleaved caspase-3 were tested by Western blotting. The technique of immunohistochemical staining was used to detect the immunoreactivity positive cells of phosphorylated (p)-Akt and cleaved caspase-3 in the spinal cord. Double staining of immunohistochemistry was used to examine the cellular location of the p-Akt and cleaved caspase-3 positive cells. RESULTS: The chronic intrathecal injection of morphine (15 μg) for 7 d markedly upregulated the spinal protein levels of p-Akt and cleaved caspase-3 in the rats. Thirty min before injection of morphine, intrathecal injection of leptin antagonist (3 μg) for 7 d significantly attenuated the upregulation of the protein levels of p-Akt and cleaved caspase-3 induced by chronic morphine treatment. The p-Akt was exclusively observed in the spinal neurons. The cleaved caspase-3 was only localized with the spinal astrocytes. Intrathecal injecting the inhibitors of leptin, Akt and caspase-3 ameliorated the chronic antinociceptive tolerance. CONCLUSION: The spinal Akt pathway and active caspase-3 are involved in the leptin-mediated chronic morphine antinociceptive tolerance in rats.     

Effect of HO-1 expression on neuronal apoptosis in spinal cord of rats with formaldehyde inflammatory pain

AIM: To observe the change of heme oxygenase(HO-1)expression in the spinal cord during formaldehyde-induced pathological pain and investigate the effect of HO-1 on neuronal apoptosis in the spinal cord induced by formaldehyde inflammatory pain.METHODS: The protein expression of HO-1 in the left and the right spinal dorsal horn was detected by Western blotting. The neuronal apoptosis rate of the spinal cord was determined by flow cytometry.RESULTS: Compared to control group, the protein expression of HO-1 was significantly up-regulated at different time points after the injection of formaldehyde, which was most obviously 24 h after the injection of formaldehyde and was still higher than that in control group at 72 h. Compared to control group, the neuronal apoptosis rate of spinal cord increased in the rats with formaldehyde inflammatory pain. No significant difference of the neuronal apoptosis rate was observed between formaldehyde group and solvent control group . Intrathecal injection of 100 μg ZnppIX, an inhibitor of HO-1, attenuated the degree of spontaneous pain response, but induced an increase in the rate of neuronal apoptosis in spinal cord in the rats with formaldehyde inflammatory pain. CONCLUSION: Formaldehyde inflammatory pain induces the increases in HO-1 expression and neuronal apoptosis in the rat spinal cord. HO-1 promotes the response of spontaneous pain and inhibits the process of neuronal apoptosis in spinal cord induced by formaldehyde inflammatory pain.     

Effect of endoplasmic reticulum stress protein BiP on type 2 diabetic neuropathic pain in rats treated with curcumin

AIM: To investigate the effects of immunoglobulin heavy chain-binding protein (BiP),an endoplasmic reticulum stress protein, on mechanical withdrawal threshold (MWT), thermal withdrawal latency (TWL), spinal dorsal horn and dorsal root ganglion (DRG) in type Ⅱ diabetic neuropathic pain rats treated with curcumin. METHODS: The rats were fed with a high-fat and high-fructose diet for 8 weeks to induce insulin resistance, and then were intraperitoneally injected with streptozotocin (STZ, 35 mg/kg). Eighty-one rats were selected into experimental design as their blood glucose ≥ 16.7 mmol/L 3 d after STZ injection and their MWT and TWL were decreased to 85% of the baseline values 14 d after STZ injection. The rats were divided into 3 groups (n=27 each): DNP group: type 2 diabetic neuropathic pain; DCur group: type 2 diabetic neuropathic pain and intraperitonal injection of curcumin at a dose of 100 mg·kg^-1·d^-1; DSC group: type 2 diabetic neuropathic pain and intraperitonal injection of corn oil at a dose of 4 mL/kg. Another 27 normal SD male rats fed with normal forage were adopted as control group (C group). MWT and TWL were measured at the time points of 3 d, 7 d and 14 d after curcumin injection. The lumbar segment 4~6 of the spinal cord and the corresponding DRG were removed at the same time. The expression of BiP was determined by immunohistochemical staining and Western blotting. RESULTS: Compared with C group, the rats in DNP group developed hyperglycemia and a decrease in MWT and TWL, as well as an increase in the activity of BiP in spinal dorsal horn and DRG (P<0.05). Compared with DNP group, the rats in DCur group at the time point of 7 d significantly attenuated mechanical allodynia and thermal hyperalgesia, and these effects were correlated with the inhibition of BiP hyper-activation at the time point of 14 d after treatment with curcumin (P<0.05). No significant difference of MWT, TWL and the expression of BiP between DNP group and SC group was observed. CONCLUSION: BiP participates in the pathogenesis of type Ⅱ diabetic neuropathic pain. Curcumin attenuates the MWT and TWL in type 2 diabetic neuropathic pain rats. The mechanism may be involved in the inhibition of BiP expression by curcumin.     

Effect of progesterone on SH-SY5Y cells injured by ATP

AIM: To investigate the neuroprotective effect of progesterone against adenosine triphosphate (ATP)-injured human neuroblastoma SH-SY5Y cells.METHODS: The SH-SY5Y cells in the logarithmic phase were divided into different groups according to the progesterone and ATP concentrations. The cell viability was measured by CCK-8 assay. The membrane permeability was detected using fluorescent dye YO-PRO-1. Cytosolic Ca²⁺ concentration was measured with fluorescent dye Fluo-3/AM. The expression of purinergic P2X₇ receptor was assessed by Western blot.RESULTS: The viability of the SH-SY5Y cells was significantly decreased (P<0.05) and YO-PRO-1 uptake was obviously increased (P<0.05) in a concentration-dependent manner compared with control group when SH-SY5Y cells were treated with ATP at 1, 3, 5 and 7 mmol/L for 2 h. The viability reduction of the SH-SY5Y cells induced by ATP was obviously counteracted by treatment with progesterone at 3, 10 and 30 nmol/L for 30 min (P<0.05) as compared with ATP group. YO-PRO-1 fluorescence enhancement induced by ATP in SH-SY5Y cells was significantly reduced (P<0.05) by progesterone (30 nmol/L) or P2X₇ receptor antagonist KN-62 (500 nmol/L) pretreatment for 30 min, and no obvious difference between treatments with progesterone and KN-62 was observed. Cytosolic Ca²⁺ fluorescence intensity in normal group was a little, but that in ATP group was increased (P<0.05). Progesterone or KN-62 pretreatment significantly decreased the cytosolic fluorescence intensity of Ca²⁺ induced by ATP (P<0.05). However, no obvious difference between treatments with progesterone and KN-62 was found. The expression of P2X₇ receptor in ATP group was significantly higher than that in control group (P<0.05), and progesterone inhibited ATP-induced P2X₇ receptor expression (P<0.05).CONCLUSION: Progesterone inhibits P2X₇ receptor expression, membrane pore formation, intracellular Ca²⁺ increase and cell death induced by ATP, so progesterone may protect SH-SY5Y cells against ATP-induced injuries.     

Expression of nuclear factor kappa B in the spinal ganglions of adjuvant arthritis rats

AIM: To study the expression of nuclear factor kappa B (NF-κB) in the L1-L4 spinal ganglions of adjuvant arthritis (AA) rats, to explore the neuroendocrinoimmunological mechanism of rheumatoid arthritis (RA). METHODS: The expression of the NF-κB protein was detected by immunohistochemistry method and the alteration of the active NF-κB protein (nuclear protein) was investigated by Western blotting analysis. RESULTS: The expression of NF-κB protein in L1-L4 spinal ganglions of AA rats was much higher than that in the healthy controls (P<001), and the active NF-κB content in L1-L4 spinal ganglions in AA rats was much higher than that in the healthy controls (P<001). CONCLUSION: The active NF-κB protein in the spinal ganglions may be involved in the pathogenesis of the rheumatoid arthritis.     

Effects of low-glucose on long-term potentiation in the hippocampal slices of immature and adult rats

AIM: The effects of low-glucose on long-term potentiation (LTP) in the CA1 region of hippocapal slices of immature (15-16 days old) and adult (56-63 days old) rats were examined. METHODS: The technique of electrophysiology was used, and the slopes of the field excitatory postsynaptic potentials (S-EPSP) were measured. RESULTS: When slices were exposed to glucose medium at concentrations of 3 or 1.5 mmol/L, S-EPSP decreased significantly. In the slices from adult rats, only short-term potentiation was elicited by high frequency stimulation in the medium of 3 or 1.5 mmol/L glucose. However, in the slices from immature rats, LTP was still induced in the medium of 3 mmol/L glucose. CONCLUSION: Low-glucose medium depressed the synaptic transmission. In terms of the synaptic plasticity, the low-glucose endurance in immature rats was stronger than that in adult rats.     

Role of autophagy in intervertebral disc degeneration in diabetic rats

AIM:To explore the levels of apoptosis and autophagy in the nucleus pulposus tissues of intervertebral discs in diabetic rats. METHODS:Sixteen weeks after injection of streptozocin (STZ), the lumbar intervertebral discs were obtained from the rats. The histological changes were observed by hematoxylin-eosin (HE) staining and alcian blue staining. The apoptosis of the nucleus pulposus cells was measured by the methods of terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL), immunohistochemistry, and Western blotting. The level of autophagy in the nucleus pulposus cells was detected by Western blotting and immunohistochemistry. RESULTS:Compared with normal group, HE and alcian blue staining suggested that the intervertebral discs of the diabetic rats became degenerate. The expression of caspase-3 and the apoptotic rate were increased in intervertebral disc nucleus pulposus of the diabetic rats. The results of immunohistochemistry and Western blotting showed that the expression levels of LC3Ⅱ/LC3Ⅰand beclin-1 in the diabetic rats were higher than those in normal group. CONCLUSION: The STZ-induced diabetes accelerates degeneration of the intervertebral discs. In addition, the apoptosis and autophagy are increased in the intervertebral discs of diabetic rats.     

Upregulation of neurokinin A levels by nerve growth factor in the experimental asthmatic guinea pig

AIM: To explore the regulatory mechanism of nerve growth factor (NGF) on neurokinin A in the experimental asthmatic guinea pig. METHODS: Radioimmunoassay was used to determine the alteration of neurokinin A levels in the lower respiratory tract and visceral sensory afferent sites while NGF was absent (inhalation of NGF antibody through nasal cavity) in the asthmatic guinea pig. RESULTS: The contents of neurokinin A in the trachea, bronchus, lung, C₇-T₅ spinal ganglia and the correspondent spinal dorsal horn, nodose ganglia and solitary nucleus area in the experimental asthmatic guinea pig with the absent of NGF in the respiratory tract were much lower than those in the asthmatic and control groups (P＜0.01). CONCLUSION: NGF upregulated the contents of neurokinin A in the lower respiratory tract and visceral sensory sites of the experimental asthmatic guinea pig, and both might be involved in the pathogenesis of asthma.     

Up-regulatory effect of nerve growth factor on substance P in the experimental asthma

AIM: To explore the regulatory mechanism of nerve growth factor (NGF) on substance P in the experimental asthmatic guinea pig. METHODS: Radioimmunoassay was used to determine the alteration of substance P levels in the lower respiratory tract and visceral sensory afferent sites while NGF was absent (inhalation of NGF antibody through nasal cavity) in the asthmatic guinea pig. RESULTS: The contents of substance P in the trachea, bronchus, lung, C7-T5 spinal ganglia and the correspondent spinal dorsal horn, nodose ganglia and solitary nucleus area in the experimental asthmatic guinea pig with the absent of NGF in the respiratory tract, were much lower than those in the asthmatic and control groups (P<0.01). CONCLUSION: NGF upregulates the contents of substance P in the lower respiratory tract and visceral sensory sites of the experimental asthmatic guinea pig, which might be involved in the pathogenesis of asthma.     

Exogenous ATP induces formation of membrane pore in PC12 cells

AIM：To investigate the formation of membrane pore in PC12 cells induced by exogenous adenosine triphosphate (ATP) and to identify the key molecular targets. METHODS：PC12 cells were treated with different concentrations of ATP to establish the injury model. The morphological change was observed under an inverted phase-contrast microscope. The viability of the PC12 cells was measured by CCK-8 assay. Fluorescent dye YO-PRO-1 was used to detect the membrane permeability. The expression of P2X7 receptor and pannexin 1 (Panx1) at mRNA and protein levels was assessed by real-time PCR and Western blotting. RESULTS：After exposed to ATP (1 mmol/L, 3 mmol/L and 5 mmol/L) for 3 h, the PC12 cells became edematous, and the number of adherent cells decreased gradually in a dose-dependent manner. The cell viabilities in 3 mmol/L ATP group and 5 mmol/L ATP group were significantly decreased compared with control group (P<0.05). YO-PRO-1 uptake in the PC12 cells exposed to ATP (0, 1, 3 and 5 mmol/L) for 15 min, 30 min and 60 min increased in a dose-dependent and time-dependent manner. The cell viability increased and the intracellular fluorescence intensity induced by ATP were significantly antagonized in brilliant blue G (a P2X7 receptor inhibitor) pretreatment group (P<0.05), whereas it did not change in carbenoxolone (a Panx1 inhibitor) pretreatment group (P>0.05). The expression of P2X7 receptor at mRNA and protein levels was significantly increased (P<0.05), but the expression of Panx1 was not changed (P>0.05) when PC12 cells were exposed to ATP for 3 h. CONCLUSION：Extracellular ATP at high concentration may induce membrane pore formation with the expression and activation of P2X7 receptor in PC12 cells.     

龙眼‘蜀冠’ב大乌圆’杂交后代果实品质性状多样性分析及综合评价

以龙眼‘蜀冠’ב大乌圆’的F1代65个单株为材料，对其果实的27个品质性状进行了连续3年的观测，应用SPSS19.0软件对各性状进行变异系数、Shannon-Weinner多样性指数和主成分分析。结果表明：65个单株果实品质性状存在着极显著差异，13个数量性状（单果质量、果实纵径、果实横径、可溶性固形物、可食率、果皮厚度、果皮质量、果肉厚、种子质量、维生素C含量、蔗糖含量、葡萄糖含量和果糖含量）变异系数在6.95% ~ 41.64%之间；多样性指数在1.33 ~ 2.02之间；14个描述性状（成熟期、果皮颜色、果形、果肩、果顶、龟裂纹、疣状突起、果肉颜色、果肉质地、果肉透明度、果肉离核难易度、果汁、化渣程度和果肉风味）多样性指数在0.56 ~ 1.28之间；主成分分析发现前5个性状（单果质量、可食率、可溶性固形物、成熟期和果实风味）的累积贡献率为87.50%；建立了‘蜀冠’ב大乌圆’杂交后代果实品质性状综合评价的方法，利用该方法在65个杂交后代中筛选出了5个优异的单株（SD7、SD11、SD36、SD53和SD120），结果与实际观测结果高度一致。     

Effect of JNK/MCP-1 signaling pathway on anti-diabetic neuropathic pain by curcumin in type 2 diabetic rats

AIM:To investigate the effect of JNK/MCP-1 signaling pathway on anti-diabetic neuropathic pain by curcumin in type 2 diabetic rats. METHODS:The male Sprague-Dawley rats were induced as the model of the type 2 diabetic neuropathic pain rats, they were randomly divided into 6 groups (n=27): type 2 diabetic neuropathic pain (DNP) group, type 2 diabetic neuropathic pain and intraperitoneal injection of curcumin (Cur) group, type 2 diabetic neuropathic pain and solvent control (DSC) group, type 2 diabetic neuropathic pain and JNK inhibitor (DJ) group, type 2 diabetic neuropathic pain and JNK inhibitor solvent control (DJS) group, type 2 diabetic neuropathic pain and monocyte chemoattractant protein 1 (MCP-1) agonist (DM) group. Another 27 normal SD rats were selected as control group. Mechanical withdrawal threshod and thermal withdrawal latency were measured at 3rd d, 7th d and 14th d after dosing, then the lumbar segment 4~6 of the spinal cord and L4~6 DRG were removed at the same time. ELISA was used to measure MCP-1 level. The expression of p-JNK was determined by Western blotting. RESULTS:Compared with DNP group, p-JNK was significantly decreased at 7th d and 14th d in Cur group, DJ group and DM group after treatment (P<0.05). Compared with C group, the MCP-1 was significantly declined in other 6 group after streptozotocin injection (P<0.05). Compared with DNP group, MCP-1 were significantly increased at 7th d and 14th d in Cur group and DJ group after treatment (P<0.05), and that in DM group was greatly decreased (P<0.05). CONCLUSION: The expression of p-JNK and MCP-1 was increased in DNP rats with spinal cord and dorsal root ganglion. The mechanism of curcumin reducing the neuropathic pain in type 2 diabetic rats might be through regulating the JNK/MCP-1 pathway.