度洛西汀对炎性疼痛大鼠海马5-羟色胺5-羟色胺2A受体基因和脑源性神经营养因子基因表达的干预作用

doi:10.3877/cma.j.issn.2095-655X.2017.02.015

目的

探讨度洛西汀对炎性疼痛大鼠海马5-羟色胺(5-HT)水平、5-HT2A受体mRNA和脑源性神经营养因子(BDNF)mRNA表达的干预作用。

方法

54只成年雄性SD大鼠随机数字表法分为A、B、和C组，每组各18只，各组处理：A组(足底注射生理盐水)、B组(炎性疼痛模型)和C组(炎性疼痛模型＋度洛西汀灌胃)。炎性疼痛模型是大鼠左侧足底皮下注射完全弗氏佐剂(CFA)和生理盐水(1∶1)混合溶剂100 μL，建立慢性炎症疼痛模型。度洛西汀[10 mg/(kg·d)]灌胃2周。在建模前及处理2周后采用电子测痛仪检测大鼠左侧足底50%缩足阈值(PWT)的变化，用糖水偏爱实验检测大鼠抑郁样行为，采用RT-PCR检测不同组间大鼠海马内5-HT2A受体mRNA和BDNF mRNA的表达，ELISA的方法检测海马中5-HT的变化。

结果

各组大鼠在处理前，50%PWT分别为[(52.60±1.60)g]、[(52.93±1.95)g]、[(51.47±2.59)g]，差异无统计学意义(F＝0.81，P＝0.46)。2周后B组的50%PWT[(41.97±3.88)g]以及糖水消耗率[(80.12±2.91)%]与A组[(49.30±3.20)g，(87.33±5.22)%]比较，差异有统计学意义(LSD-t＝2.46，3.21；P＝0.03，0.01)。C组50%PWT[(49.83±7.39)g]和糖水消耗率[(85.52±3.13)%]与B组相比升高(LSD-t＝2.64，2.40；P＝0.02，0.03)，C组与A组的痛阈(LSD-t＝0.17，P＝0.86)及糖水消耗率(LSD-t＝0.44，P＝0.80)相比较，差异无统计学意义。B组与A组相比较，海马中的5-HT[(1.69±0.26)ng/mg；(2.27±0.30)ng/mg]降低(LSD-t＝2.38，P＝0.03)，海马中5-HT2A受体mRNA(0.62±0.28；1.20±0.30)差异无统计学意义(LSD-t＝1.57，P＝0.14)，BDNF mRNA(0.68±0.29；1.58±0.60)的表达降低(LSD-t＝2.46，P＝0.03)。C组海马中的5-HT[(2.82±0.62)ng/mg]与B组比较上升(LSD-t＝4.61，P＝0.00)，C组与A组相比较海马中的5-HT上升(LSD-t＝2.23，P＝0.04)。同时C组海马中5-HT2A受体mRNA(1.70±1.04)和BDNF mRNA(1.75±0.88)的表达都是升高的(LSD-t＝2.91，2.91；P＝0.01，0.01)，C组与A组的5-HT2A受体(LSD-t＝1.34，P＝0.200)及BDNF mRNA(LSD-t＝0.45，P＝0.65)表达相比较，差异无统计学意义。

结论

慢性炎性疼痛大鼠会出现痛阈下降和抑郁样行为变化，同时海马内5-HT含量和BDNF mRNA的表达下降；度洛西汀可以提高慢性炎性疼痛大鼠的痛阈，改善抑郁样行为，其机制可能通过海马中5-HT含量、5-HT2A受体mRNA和BDNF mRNA的表达升高发挥作用。

关键词: 炎性疼痛, 海马, 5-羟色胺, 脑源性神经营养因子, 痛阈

Objective

To explore the changes of depressive-like behavior, pain threshold and the expression of 5-HT and 5-HT2A receptor mRNA and BDNF mRNA in hippocampus of rats with inflammatory pain and the intervention of duloxetine, and study the relationship between nerve material change in hippocampus and inflammatory pain.

Methods

Fifty-four adult male SD rats were divided into A, B and C group by random number table, 18 rats in each group.Processing of each groups: group A (plantar injection of saline solution), B group (inflammatory pain model), C group(inflammatory pain model+ duloxetine by intragastric administration). The inflammatory pain models were established by subcutaneously injecting mixture of CFA and saline into the left plantar of the rats.Duloxetine [10 mg/(kg·d)] was delivered to rats by intragastric administration for 2 weeks.The alterations of 50% paw withdrawal threshold (PWT) were measured before and 2 weeks after CFA injection with Von Frey Fibres.The depression-like behaviors tested with sucrose preference test.While the levels of 5-HT2A receptor mRNA and BDNF mRNA in the hippocampus were detected by RT-PCR and the expression of 5-HT in the hippocampus was detected by ELISA.

Results

Before treatment, the 50% PWT of the three groups were [(52.60±1.60)g], [(52.93±1.95)g], [(51.47±2.59)g], the difference had no statistical significance (F=0.81, P=0.46). Two weeks later, 50%PWT [(41.97±3.88)g] and sugar consumption rate [(80.12±2.91)%] of the group B lower than those of the group A [(49.30±3.20)g, (87.33±5.22)%], the differences had statistical significance (LSD-t=2.46, 3.21; P=0.03, 0.01). Compared with the group B, the 50%PWT [(49.83±7.39)g] and sugar consumption rate [(85.52±3.13)%] in the group C were higher [LSD-t=2.64, 2.40; P=0.02, 0.03]. Compared with A group, 5-HT of B group [(1.69±0.26)ng/mg; (2.27±0.30)ng/mg] in hippocampus was decreased (LSD-t=2.38, P=0.03), the expressions of 5-HT2A receptor mRNA of the two groups (0.62±0.28; 1.20±0.30) in the hippocampus had no statistical significance (LSD-t=1.57, P=0.14), the expression of BDNF mRNA of group B (0.68±0.29; 1.58±0.60) was lower (LSD-t=2.46, P=0.03). Compared with group B, the 5-HT of group C [(2.82±0.62)ng/mg] in hippocampus was increased (LSD-t=4.61, P=0.00), the expressions of 5-HT2A receptor mRNA (1.70±1.04) and BDNF mRNA (1.75±0.88) of group C in hippocampus were higher (LSD-t=2.91, 2.91; P=0.01, 0.01).

Conclusions

The pain threshold decrease and depression-like behavior appears, and the expression of 5-HT, 5-HT2A receptor mRNA and BDNF mRNA in hippocampus decrease in the rat with chronic inflammatory pain.Duloxetine can improve pain threshold and depression-like behavior of the rat with chronic inflammatory pain by increasing the expression of 5-HT, 5-HT2A receptor mRNA and the BDNF mRNA in hippocampus.

Key words: Inflammatory pain, Hippocampus, 5-hydroxytryptamine, Brain-derived neurotrophic factor, Pain threshold

表1 各组大鼠50%缩足痛阈值和糖水消耗率(±s)

组别	鼠数(只)	造模2周后痛阈(g)	糖水消耗率(%)
A组	6	49.30±3.20	87.33±5.22
B组	6	41.97±3.88^a	80.12±2.91^a
C组	6	49.83±7.39^b	85.52±3.13^b
F值	?	4.36	5.57
P值	?	0.03	0.02

表1 各组大鼠50%缩足痛阈值和糖水消耗率(±s)

组别	鼠数(只)	造模2周后痛阈(g)	糖水消耗率(%)
A组	6	49.30±3.20	87.33±5.22
B组	6	41.97±3.88^a	80.12±2.91^a
C组	6	49.83±7.39^b	85.52±3.13^b
F值	?	4.36	5.57
P值	?	0.03	0.02

表2 各组大鼠海马中5-HT2A mRNA及BDNF mRNA的表达(±s)

组别	鼠数(只)	5-HT2A mRNA	BDNF mRNA
A组	6	1.20±0.3	1.58±0.60
B组	6	0.62±0.28	0.68±0.29^a
C组	6	1.70±1.04^b	1.75±0.88^b
F值	?	4.25	4.91
P值	?	0.04	0.02

表2 各组大鼠海马中5-HT2A mRNA及BDNF mRNA的表达(±s)

组别	鼠数(只)	5-HT2A mRNA	BDNF mRNA
A组	6	1.20±0.3	1.58±0.60
B组	6	0.62±0.28	0.68±0.29^a
C组	6	1.70±1.04^b	1.75±0.88^b
F值	?	4.25	4.91
P值	?	0.04	0.02

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Intervention of duloxetine in the expression of 5-HT and 5-HT2A receptor mRNA and BDNF mRNA in hippocampus of rats with inflammatory pain

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Intervention of duloxetine in the expression of 5-HT and 5-HT2A receptor mRNA and BDNF mRNA in hippocampus of rats with inflammatory pain