| 邵深深,张蔷蓉,林希,等.苍耳子水提液对过敏性鼻炎小鼠的治疗作用及机制研究[J].浙江中医药大学学报,2023,47(2):111-117. |
| 苍耳子水提液对过敏性鼻炎小鼠的治疗作用及机制研究 |
| Study on the Therapeutic Effect and Mechanism of Xanthi Fructus Water Extract on Mice with Allergic Rhinitis |
| DOI：10.16466/j.issn1005-5509.2023.02.001 |
| 中文关键词: 苍耳子水提液 过敏性鼻炎 肥大细胞脱颗粒 组胺 LAT/PLCγ1/PKC信号通路 作用机制 |
| 英文关键词: Xanthii Fructus water extract allergic rhinitis mast cell degranulation histamine LAT/PLCγ/PKC signaling pathway functional mechanism |
| 基金项目:温州市科技计划项目(Y20180817) |
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| 摘要点击次数: 1979 |
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| 中文摘要: |
| [目的] 探讨苍耳子(Xanthii Fructus，XF)水提液对过敏性鼻炎(allergic rhinitis，AR)小鼠的治疗作用及机制。[方法] 36只BALB/c小鼠随机分为正常对照(normal control，NC)组、AR组、低剂量XF水提液(low-dose XF，L-XF)组(4 mg·kg-1)、中剂量XF水提液(medium-dose XF，M-XF)组(8 mg·kg-1)、高剂量XF水提液(high-dose XF，H-XF)组(16 mg·kg-1)和地塞米松(dexamethasone group，DXM)组(5 mg·kg-1)，每组6只。除NC组外，余下各组小鼠均用卵清白蛋白(ovalbumin，OVA)致敏进行AR造模，随后治疗14 d。治疗结束后，记录小鼠搔鼻和喷嚏次数，并进行鼻炎症状评分；取小鼠鼻腔组织，行苏木精-伊红(hematoxylin-eosin，HE)染色和糖原(periodic acid-schiff，PAS)染色观察小鼠鼻腔组织学形态变化并评分，中性红染色观察肥大细胞脱颗粒情况，并计算各组药物脱颗粒抑制率；酶联免疫吸附试验检测小鼠血清白三烯C4、组胺及β-氨基己糖苷酶A水平；免疫印迹法检测鼻腔组织T细胞激活连接蛋白(linker for activation of T cell，LAT)、磷酸化LAT(phospho-LAT，p-LAT)、磷脂酶Cγ1(phospholipase Cγ1，PLCγ1)、磷酸化PLCγ1(phospho-PLCγ1，p-PLCγ1)、蛋白激酶C(protein kinase C，PKC)、磷酸化PKC(phospho-PKC，p-PKC)蛋白的表达。 [结果] 与NC组比较，AR组小鼠的搔鼻、打喷嚏次数显著增加，鼻炎评分显著升高，HE评分以及PAS评分均显著升高，血清白三烯C4、组胺、β-氨基己糖苷酶A水平显著升高，肥大细胞脱颗粒率显著升高，p-LAT/LAT、p-PLCγ1/PLCγ1、p-PKC/PKC的蛋白表达显著升高(P＜0.01)。与AR组比较，各组小鼠搔鼻、打喷嚏次数，以及肥大细胞脱颗粒率显著减少(P＜0.05，P＜0.01)；M-XF组、H-XF组、DXM组的鼻炎评分显著下降，HE评分以及PAS评分显著降低，血清白三烯C4、组胺、β-氨基己糖苷酶A水平显著下降(P＜0.05，P＜0.01)；M-XF组、H-XF组、DXM组的药物脱颗粒抑制率显著高于L-XF组(P＜0.05，P＜0.01)。免疫印迹显示，与AR组比较，各组小鼠鼻腔组织的p-PKC/PKC蛋白表达显著下降(P＜0.05，P＜0.01)，M-XF组、H-XF组、DXM组小鼠鼻腔组织的p-LAT/LAT、p-PLCγ1/PLCγ1的蛋白表达显著下降(P＜0.05，P＜0.01)。 [结论] XF水提液可能通过抑制LAT/PLCγ1/PKC信号通路的表达，抑制肥大细胞脱颗粒，从而改善小鼠AR症状。 |
| 英文摘要: |
| [Objective] To investigate the therapeutic effect and mechanism of Xanthii Fructus(XF) water extract on allergic rhinitis(AR) mice. [Methods] Thirty-six BALB/c mice were randomly divided into normal control(NC) group， AR model(AR) group， low dose XF water extract(L-XF) group(4 mg·kg-1)， medium dose XF water extract(M-XF) group(8 mg·kg-1)， high dose XF water extract(H-XF) group(16 mg·kg-1)， and dexamethasone(DXM) group(5 mg·kg-1)， six in each group. Except the NC group， mice were sensitized with ovalbumin(OVA) to establish AR model， then treated for 14 days. After treatment， the times of nasal scratching and sneezing were recorded and the symptoms of rhinitis were scored. Mice nasal tissues were collected， hematoxylin-eosin(HE) staining and periodic acid-schiff(PAS) staining were used to observe the histological changes and scoring， neutral red staining was used to count the number of degranulation of mast cells and calculate the drug degranulation inhibition rate； enzyme linked immunosorbent assay(ELISA) was used to detect serum leukotriene C4(LTC4)， histamine and β-hexosaminidase A levels； Western blot was used to detect protein expression of linker for activation of T cell(LAT)， phospho-LAT(p-LAT)， phospholipase Cγ1(PLCγ1)， phospho-PLCγ1(p-PLCγ1)， protein kinase C(PKC)， phospho-PKC(p-PKC) in nasal tissue. [Results] Compared with NC group， the times of scratching nose and sneezing， rhinitis score， HE and PAS scores， serum LTC4， histamine and β-hexosaminidase A levels， rate of degranulation mast cells， and protein expression of p-LAT/LAT， p-PLCγ1/PLCγ1， p-PKC/PKC in AR group were significantly increased(P＜0.01). Compared with AR group， the times of scratching nose and sneezing and rate of degranulation mast cells in each group were significantly reduced(P＜0.05， P＜0.01)； and rhinitis score， HE and PAS scores， serum LTC4， histamine and β-hexosaminidase A levels in M-XF group， H-XF group and DXM group were significantly decreased(P＜0.05， P＜0.01)； the drug degranulation inhibition rate of M-XF group， H-XF group and DXM group was significantly higher than that of L-XF group(P＜0.05， P＜0.01). Western blot showed that compared with AR group， protein expression of p-PKC/PKC was significant decreased in each group(P＜0.05， P＜0.01)， and protein expression of p-LAT/LAT、p-PLCγ1/PLCγ1 were significantly decreased in M-XF group， H-XF group and DXM group(P＜0.05， P＜0.01). [Conclusion] XF water extract may ameliorate mice AR symptom mediated inhibition of LAT/PLCγ1/PKC signaling pathway to restrain mast cell degranulation. |
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