青光安Ⅱ号方通过Brn-3b调控Bax/Bcl-2/Caspase-3抑制青光眼RGCs凋亡的机制研究  

Mechanism of QingguanganⅡFormula Inhibiting RGCs Apoptosis in Glaucoma via Brn-3b/Bax/Bcl-2/Caspase-3 Regulation

作　　者：李文娟 谭涵宇 时健 彭榆津 周蓓 彭清华 LI Wen-juan;TAN Han-yu;SHI Jian;PENG Yu-jin;ZHOU Bei;PENG Qing-hua(Department of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208;Hunan Provincial Key Laboratory of Ophthalmolog,Otorhinolaryngology and Chinese Medicine,Changsha 410208;Department of Ophthalmology,Yueyang Hospital Affiliated to Hunan University of Chinese Medicine,Hunan 414000)

机构地区：[1]湖南中医药大学中西医结合学院,长沙410208 [2]中医药防治眼耳鼻喉疾病湖南省重点实验室,长沙410208 [3]湖南中医药大学附属岳阳医院眼科,湖南414000

出　　处：《中国中西医结合杂志》2025年第3期330-336,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上基金(No.82274588);湖南省中医药科研课题(No.B2023093)。

摘　　要：目的探讨青光安Ⅱ号方通过脑特定蛋白3B(Brn-3b)调控B淋巴细胞瘤-2基因(Bcl-2)/Bcl-2相关X蛋白(Bax)/半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)抑制青光眼RGCs凋亡的机制。方法将60只SPF级健康雄性SD大鼠,饲养1周后使用随机数字表将大鼠分入空白组、模型组、青光安Ⅱ号方低剂量组、青光安Ⅱ号方中剂量组、青光安Ⅱ号方高剂量组和益脉康组,每组10只。除空白组,其余各组烙闭巩膜上静脉引起大鼠眼压升高造模。空白组和模型组用生理盐水灌胃,益脉康组和青光安Ⅱ号方各剂量组分别用益脉康[9.67 g/(kg·d)]和相应剂量青光安Ⅱ号方[3.375、6.75、13.5 g/(kg·d)]灌胃,每组均灌胃4周。术前、术后30 min、术后1、7、14、28天各组测量眼压。处死大鼠后分离视网膜组织,部分组织在光镜下观察,并检测每组大鼠视网膜RGCs凋亡以及RGCs中Brn-3b、Caspase-3、Bcl-2、Bax蛋白及mRNA的表达情况。结果各造模组大鼠眼压均较造模前有所升高(P<0.05),且在4周时也能保持高眼压状态。与同时期模型组比较,青光安Ⅱ号方各组和益脉康组RGCs凋亡数量均明显降低(P<0.01);与同时期青光安Ⅱ号方低、中剂量组和益脉康组比较,青光安Ⅱ号方高剂量组RGCs凋亡数量最低(P<0.05)。与模型组比较,益脉康组与青光安Ⅱ号方高剂量组Caspase-3和Bax蛋白及mRNA表达明显升高(P<0.01),益脉康组、青光安Ⅱ号方高剂量组Brn-3和Bcl-2蛋白及mRNA表达明显降低(P<0.01)。结论青光安Ⅱ号方通过调控Brn-3b及Bax/Bcl-2/Caspase-3来抑制RGCs凋亡与,与益脉康疗效相当。Objective To explore the mechanism by which QingguanganⅡFormula(QGAⅡ)inhibits the apoptosis of retinal ganglion cells(RGCs)in glaucoma through the regulation of B-cell lymphoma 2(Bcl-2)/Bcl-2-associated X protein(Bax)/Cysteine-aspartic protease 3(Caspase-3)by Brain-specific homeobox/POU domain protein 3B(Brn-3b).Methods After one week of acclimatization sixty healthy male SPF SD rats were randomly divided into control group,model group,low-dose,medium-dose,and high-dose QGAⅡgroup,and Yimaikang group,10 in each group.Except for the control group,elevated intraocular pressure was induced in the other groups by cauterizing the episcleral veins.The control and model groups were treated with saline,while the Yimaikang(9.67 g·kg-1·d-1)and QGAⅡ(3.375,6.75,13.5 g·kg-1·d-1)groups received their respective treatments via gavage for 4 weeks.Intraocular pressure was measured preoperatively and at 30minutes,1,7,14,and 28 days postoperatively.After euthanizing the rats,retinal tissues were isolated for histological examination and analysis of RGC apoptosis and the expression of Brn-3b,Caspase-3,Bcl-2,and Bax proteins and mRNA.Results Intraocular pressure was significantly elevated in all model groups compared to baseline(P<0.05)and remained high at 4 weeks.Compared to the model group,the QGAⅡand Yimaikang groups exhibited significantly reduced RGC apoptosis(P<0.01).Among them,the high-dose QGAⅡgroup showed the lowest RGC apoptosis rates(P<0.05).Compared to the model group,expressions of Caspase-3and Bax proteins and mRNA increased in the Yimaikang and high-dose QGAⅡgroups(P<0.01),while expressions of Brn-3b and Bcl-2 proteins and mRNA significantly decreased(P<0.01).Conclusion QGAⅡinhibits RGC apoptosis by regulating Brn-3b,Bax/Bcl-2/Caspase-3,with efficacy comparable to Yimaikang.

关 键 词：青光安Ⅱ号方 脑特定蛋白3B B淋巴细胞瘤-2基因/Bcl-2相关X蛋白/半胱氨酸天冬氨酸蛋白酶-3 视网膜神经节细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学]