Acupuncture promotes tear secretion by up-regulating VIP/cAMP/PKA/AQP5 signaling in guinea pigs with aqueous tear deficiency dry eye.

Xue Liu, Ying Li, Hu-Xing Shen, Wei-Ping Gao, Nan Zhao, Liu-Jiao Li, Cheng-Yong Liu
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Abstract

Objectives: To observe the effect of acupuncture on the ocular surface symptoms and the protein expression of vasoactive intestinal peptide (VIP) / cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) / aquaporin 5(AQP5) signaling pathway in lacrimal gland tissue of aqueous tear deficiency (ATD) type dry eye model, so as to investigate its mechanism underlying improvement of ATD.

Methods: British shorthair guinea pigs were randomly divided into blank control, model, acupuncture, sham-acupuncture and medication group, with 8 guinea pigs in each group. The ATD model was established by subcutaneous injection of scopolamine hydrobromide (0.6 mg/dose, 4 times/d for 10 days). For guinea pigs of the acupuncture group, filiform needles were inserted into bilateral "Jingming"(BL1), "Cuanzhu"(BL2), "Sizhukong"(TE23), "Taiyang"(EX-HN5), and "Tongziliao"(GB1) for 15 min. For guinea pigs of the sham-acupuncture group, a blunt filiform needle was used to repeatedly prick (not pierce) the skin of the same acupoints mentioned above. The treatment in the above two groups was conducted once daily for 14 days. The guinea pigs in the medication group received administration of sodium hyaluronate eye drops in both eyes, three times a day for 14 days. The objective tests of tear film break-up time (BUT), corneal fluorescein staining score (FLS) and phenol red thread (PRT) test were conducted before and after modeling and after the intervention. After the intervention, the lacrimal index (weight of lacrimal gland/body weight) was calculated. Histopathological changes of the lacrimal gland were observed after H.E. staining. The expression of AQP5 in the lacrimal gland were detected by immunofluorescence, and the contents of VIP and AQP5 in the lacrimal gland were measured by ELISA, the protein expression levels of VIP, cAMP, PKA, p-PKA and AQP5 in the lacrimal gland were detected by Western blot.

Results: In comparison with the blank control group, the PRT, BUT, lacrimal index, AQP5 immunoactivity, contents of VIP and AQP5, and protein expression levels of VIP, cAMP, PKA, p-PKA and AQP5 were significantly decreased(P<0.01, P<0.05), and FLS was obviously increased (P<0.01) in the model group . Compared to the model group, the PRT, BUT, lacrimal index, AQP5 immunoactivity, contents of VIP and AQP5, and expression levels of VIP and AQP5 in both acupuncture and medication groups, and the expression levels of cAMP, PKA, p-PKA in the acupuncture group were considerably increased (P<0.01, P<0.05), while the FLS was markedly decreased in both acupuncture and medication groups (P<0.01, P<0.05). Compared with the medication group, the acupuncture group had increased PRT (P<0.05).

Conclusions: Acupuncture intervention is effective in reducing ocular surface damage and promoting tear secretion in guinea pigs with ATD, which may be related to its function in activating VIP/cAMP/PKA signaling, and promoting the expression of AQP5 in the lacrimal gland.

针刺通过上调水性泪液缺乏性干眼症豚鼠的VIP/cAMP/PKA/AQP5信号传导来促进泪液分泌。
目的:观察针刺对水性泪液缺乏(ATD)型干眼模型泪腺组织血管活性肠肽(VIP)/环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/水通道蛋白5(AQP5)信号通路蛋白表达及眼表症状的影响,方法:将英国短毛豚鼠随机分为空白对照组、模型组、针刺组、假针刺组和药物组,每组8只。通过皮下注射氢溴酸东莨菪碱(0.6mg/剂量,4次/d,持续10天)建立ATD模型。针刺组豚鼠将丝状针插入双侧“精明”(BL1)、“宽竹”(BL2)、“四柱孔”(TE23)、“太阳”(EX-HN5)和“桐梓寮”(GB1)15分钟。假针刺组豚鼠用钝性丝状针反复刺(不刺)上述相同穴位的皮肤。上述两组的治疗每天进行一次,持续14天。药物组的豚鼠接受透明质酸钠滴眼液的双眼给药,每天三次,持续14天。在建模前后和干预后进行泪膜破裂时间(BUT)、角膜荧光素染色评分(FLS)和苯酚红线(PRT)测试的客观测试。干预后,计算泪腺指数(泪腺重量/体重)。H.E染色后观察泪腺组织病理学变化。免疫荧光法检测AQP5在泪腺中的表达,ELISA法检测VIP和AQP5的含量,Western blot法检测VIP、cAMP、PKA、p-PKA和AQP5的蛋白表达水平。结果:与空白对照组比较,PRT、BUT、泪道指数、AQP5免疫活性、VIP和AQP5含量以及VIP、cAMP、PKA、,p-PKA和AQP5显著降低(PPPPPP结论:针刺干预能有效减轻ATD豚鼠眼表损伤,促进泪液分泌,这可能与其激活VIP/cAMP/PKA信号传导,促进AQP5在泪腺的表达有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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