摘要
目的:探讨柴胡桂枝汤(CHGZT)减轻三硝基苯磺酸(TNBS)诱导的大鼠慢性胰腺炎(CP)神经性腹痛的初步机制。方法:50只雄性SD大鼠随机均分为5组,分别为假手术组、模型组、CHGZT低、中、高剂量组(4、8、16 g·kg^(-1))。假手术组轻轻翻动胰腺后即关腹,模型组采用向胰管内逆行注射2%TNBS-10%乙醇。CHGZT在造模后4周开始灌胃给药,连续2周。5组大鼠分别于术后6周进行Von Frey疼痛阈值检测;苏木素-伊红(HE)染色评估胰腺组织慢性炎症及纤维化病变;免疫组化法(IHC)检测PGP9.5蛋白表达,同时观察神经周围炎性改变;IHC及免疫荧光(IF)染色观察胰腺相关胸段脊髓后角离子钙结合蛋白-1(Iba-1)、嘌呤能受体P2X7(P2RX7)的定位及P2RX7和小胶质细胞的共表达。结果:与假手术组比较,模型大鼠胰腺组织腺泡萎缩评分、炎性浸润评分、纤维化评分显著增高(P<0.01),大鼠在不同力值疼痛反应值均明显升高(P<0.05,P<0.01),PGP9.5周围炎性浸润评分显著升高(P<0.01),模型组大鼠脊髓后角Iba-1与P2RX7表达均显著增强(P<0.01);与模型组比较,CHGZT高、中剂量组明显降低大鼠胰腺组织腺泡萎缩评分、炎性浸润评分、纤维化评分,明显降低疼痛反应值,明显降低PGP9.5周围炎性浸润评分(P<0.05,P<0.01),CHGZT高、中、低剂量组显著降低Iba-1及P2RX7表达(P<0.01)。结论:CHGZT可明显减轻CP模型大鼠腹痛程度,可能和通过抑制胰腺内神经炎症、降低相应脊髓后角小胶质细胞P2RX7活化有关。
Objective:To explore the mechanism of Chaihu Guizhitang(CHGZT)in alleviating neuropathic abdominal pain induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS)in rats with chronic pancreatitis(CP).Method:Fifty male SD rats were randomly assigned into five groups:sham operation,CP model,and low-,medium-,and high-dose(4,8,and 16 g·kg^(-1),respectively)CHGZT groups.In the sham operation group,the abdomen was closed after the pancreas was gently stirred.The rat model of CP was established by retrograde injection of 2%TNBS-10%ethanol into the pancreatic duct.The oral administration of CHGZT started 4 weeks after modeling and lasted for 2 weeks.Pain threshold was measured by Von Frey fibers 6 weeks after surgery.Hematoxylin-eosin(HE)staining was employed to reveal the chronic inflammation and fibrosis of the pancreatic tissue.Immunohistochemmistry(IHC)was employed to detect the expression of PGP9.5(a marker of pancreatic nerves)and reveal the inflammatory changes around the nerves.IHC and immunofluorescence(IF)were used to determine the location of ionized calcium-binding adaptor molecule-1(Iba-1,microglia marker)and purinergic receptor P2X7(P2RX7)and the co-expression of P2RX7 and Iba-1 in the thoracic spinal dorsal horn.Result:Compared with the sham operation group,the modeling increased the scores of pancreatic gland atrophy,inflammatory infiltration,and fibrosis(P<0.01),the abdominal pain response under different force values(P<0.05,P<0.01),and the score of peripancreatic inflammation.Moreover,the modeling up-regulated the expression of Iba-1 and P2RX7 in the thoracic spinal dorsal horn(P<0.01).Compared with the model group,the high-and medium-dose CHGZT lowered the scores of pancreatic gland atrophy,inflammatory infiltration,and fibrosis,the abdominal pain response,and the score of peripancreatic inflammation(P<0.05,P<0.01).The high-,medium-,and low-dose CHGZT all down-regulated the expression of Iba-1 and P2RX7(P<0.01).Conclusion:CHGZT can significantly relieve abdominal pain in CP rat by suppressing the inflammation around nerves in the pancreas and the P2RX7 activation of microglia in the spinal dorsal horn.
作者
李赛楠
张桂贤
沈洪昇
王曼雪
李锡晶
李霞
李文畅
肖懿
刘洪斌
LI Sainan;ZHANG Guixian;SHEN Hongsheng;WANG Manxue;LI Xijing;LI Xia;LI Wenchang;XIAO Yi;LIU Hongbin(Graduate School,Tianjin Medical University,Tianjin 300070,China;Tianjin Institute of Medical and Pharmaceutical Sciences,Tianjin 300020,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第22期40-46,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
天津市卫健委、天津市中医药管理局中医中西医结合科研课题(2021165)
天津市科技计划项目(21JCZDJC01220)。
作者简介
第一作者:李赛楠,在读硕士,从事慢性胰腺炎药理学研究,E-mail:1015740473@qq.com;通信作者:刘洪斌,博士,研究员,从事中西医结合治疗肝胆胰疾病的药理学研究,E-mail:jtss@sina.com;通信作者:张桂贤,助理研究员,从事急、慢性胰腺炎的药理学研究,E-mail:zhangguixian2007@aliyun.com。
