伏红颖,潘艳伶,陈俞如,陈洪民
FU Hongying,PAN Yanling,CHEN Yuru,CHEN Hongming

• 1:贵州医科大学临床医学院中医学教研室
• 2:贵州医科大学附属医院中医科

摘要(Abstract)：

目的 分析糖通饮含药血清对高糖环境下大鼠肾小球系膜细胞（HBZY-1）增殖的影响及可能作用机制。方法 将30只SPF级雄性SD大鼠随机均分为空白血清组和糖通饮含药血清组，糖通饮含药血清组大鼠予以糖通饮灌胃，空白血清组大鼠予以等体积的双蒸水灌胃，灌胃7 d后股动脉取血；常规培养第3～15代HBZY-1细胞分为正常糖+空白血清组、高糖+空白血清组以及高糖+糖通饮含药血清组，按分组给予相应大鼠血清干预；细胞培养结束后，CCK-8法检测各组细胞增殖情况，实时荧光定量PCR法检测各组细胞转化生长因子-β1(TGF-β1)、Smad2、Smad3 mRNA表达，Western blot法检测各组细胞TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达。结果 与正常糖+空白血清组相比，高糖+空白血清组HBZY-1细胞明显增殖，细胞中TGF-β1、Smad2、Smad3 mRNA表达增加，TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达也增加，差异均具有统计学意义（P<0.05）；与高糖+空白血清组相比，高糖+糖通饮含药血清组HBZY-1细胞增殖被抑制，细胞中的TGF-β1、Smad2、Smad3 mRNA表达降低，TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达也降低，差异均具有统计学意义（P<0.05）。结论 糖通饮可以抑制高糖状态下HBZY-1细胞的增殖，其作用机制可能与抑制TGF-β1/Smad信号通路的过度激活有关。
Objective To analyze the effect of tangtongyin drug-containing serum serum on the proliferation of rat glomerular mesangial(HBZY-1) cells in high glucose environment and its possible mechanism.Methods Thirty SPF male SD rats were randomly divided into blank serum group and tangtongyin drug-containing serum group. Rats in tangtongyin drug-containing group were fed with tangtongyin by intragastric administration, and rats in blank serum group were given equal volume double distilled water. Blood was collected from femoral artery after continuous intragastric administration for 7 days. HBZY-1 cells cultured from 3rdto 15thgeneration were divided into normal glucose + blank serum group, high glucose + blank serum group and high glucose + tangtongyin drug-containing serum group. At the end of cell culture, the cell proliferation of each group was detected by CCK-8 method, the expression ofTGF-β1,Smad2, andSmad3 mRNA was detected by real-time fluorescence quantitative PCR, and the expression of TGF-β1, p-Smad2/Smad2, and pSmad3/Smad3 protein was detected by Western blot method.Results Compared with the normal glucose + blank serum group, cells in the high glucose + blank serum group proliferated significantly, and the expression ofTGF-β1,Smad2, andSmad3 mRNA and the expression of TGF-β1, p-Smad2/Smad2, and p-Smad3/Smad3 protein were also increased(P< 0. 05). Compared with the high glucose + blank serum group, the cell proliferation was inhibited, and the expression of TTGF-β1,Smad2, andSmad3 mRNA and the protein expression of TGF-β1, p-Smad2/Smad2, and p-Smad3/Smad3 were decreased in the high glucose + tangtongyin group(P< 0. 05).Conclusion Tangtongyin can inhibit the proliferation of hbzy-1 cells in high glucose state, and its mechanism may be related to the inhibition of TGF-β1/Smad signaling pathway.

关键词(KeyWords)： 肾小球系膜细胞;细胞增殖;糖尿病肾病;大鼠;转化生长因子-β1/Smad信号通路;高糖环境;糖通饮含药血清
glomerular mesangial cells;cell proliferation;diabetic kidney disease(DKD);rat;TGF-β1/Smad signal pathway;high glucose environment;tangtongyin-containing serum

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金（82060837）;; 贵州医科大学附属医院2020年国家自然科学基金培育项目（gy5ynsfc[2020]-3）

作者(Author): 伏红颖,潘艳伶,陈俞如,陈洪民
FU Hongying,PAN Yanling,CHEN Yuru,CHEN Hongming

DOI: 10.19367/j.cnki.2096-8388.2022.06.001

参考文献(References)：

• [1]林汉英，谢乃强，李晓莉，等．肇庆市区40岁以上人群糖尿病、糖尿病前期流行病学调查[J]．广东医学，2016,37(2):285-287.
• [2]张圣英，尹长江，刘喜纲，等．中医药治疗糖尿病肾脏纤维化研究进展[J]．中国实验方剂学杂志，2022,28(11):275-282.
• [3] LV W, BOOZ G W, WANG Y, et al. Inflammation and renal fibrosis:recent developments on key signaling molecules as potential therapeutic targets[J]. Eur J Pharmacol,2018,820:65-76.
• [4]李娜，王风云，高小玲，等．中药三七防治肾纤维化的研究进展[J]．中成药，2020,42(11):2974-2978.
• [5] CHING-CHUAN H, LIN C L, HE J T, et al. Administration of cytokine-induced myeloid-derived suppressor cells ameliorates renal fibrosis in diabetic mice[J]. Stem Cell Research&Therapy,2018,9(1):183-197.
• [6]魏丹丹，莫楠，朱星昊，等．基于TGF-β1/Smad通路探讨红花黄色素对腺嘌呤致肾间质纤维化大鼠的保护作用[J]．中药新药与临床药理，2021,32(10):1437-1443.
• [7]叶斌，买鹏宇，陆智华，等．中医药干预TGF-β1/Smads信号通路对组织器官细胞纤维化调控作用的研究进展[J]．中华中医药学刊，2019,37(3):611-614.
• [8]李颖，张振，赵博，等．异丙酚调控TGF-β1/smads信号通路对UUO诱导的肾纤维化小鼠的保护作用[J]．解剖科学进展，2020,26(6):664-667,672.
• [9] AL-RASHEED N M, AL-RASHEED N M, AL-AMIN M A, et al. Fenofibrate attenuates diabetic nephropathy in experimental diabetic rat's model via suppression of augmented TGF-betal/Smad3 signalig pathway[J]. Arch Physiol Biochem,2016,122(4):186-194.
• [10] ZHAO T T, ZHANG H J, LU X G, et al. ChaihuangYishen granule inhibits diabetic kidney disease in rats through blocking TGF-β/Smad3 signaling[J]. PLoS One,2014,9(3):e90807.
• [11]房勃龙，韩鸿玲，张鹏，等．TGF-β、Smad信号通路蛋白及α-SMA在糖尿病肾病患者肾组织中的表达和意义[J]．天津医药，2013,41(11):1067-1069,1138.
• [12]高楠楠，侯胜开，潘艳伶，等．糖通饮通过抑制TGF-β1/Smad信号通路保护早期糖尿病肾病大鼠肾组织[J]．贵州医科大学学报，2019,44(1):53-58.
• [13]潘艳伶，侯胜开，高楠楠，等．糖通饮治疗早期糖尿病肾病临床研究[J]．中医药信息，2019,36(1):91-94.
• [14]钱洁玉，桑锋，刘真，等．益艾康含药血清对人NK-92MI细胞凋亡及其受体NKG2A、NKG2D、细胞因子IFN-γ表达的影响[J]．中华中医药杂志，2021,36(11):6756-6759.
• [15]王增四，高文，陈丹，等．黄芪甲苷抑制内质网应激及CHOP信号通道改善衣霉素诱导的系膜细胞凋亡[J]．医药导报，2022,41(2):150-154.
• [16]赵婷，张祥贵．TGF-β1/Smad信号通路与糖尿病肾病关系的研究进展[J]．海南医学，2021,32(5):642-646.
• [17]HUANG H, HU L, LIN J, et al. Effect of fosinopril on chemerinand VEGF expression in diabetic nephropathy rats[J]. Int J Clin Exp Pathol, 2015, 8(9):11479-11474.
• [18]凌光辉，唐文彬，孙林，等．Smad锚着蛋白在高糖诱导人肾小管上皮细胞细胞外基质沉积中的作用[J]．肾脏病与透析肾移植杂志，2012,21(4):346-352.
• [19] YANG J, ZENG Z, WU T, et al. Emodin attenuates high glucose-induced TGF-β1 and fbronectin expression in mesangial cells through inhibiton of NF-κB pathway[J]. Exp Cell Res,2013,319(20):3182-3189.
• [20]娄凯，何永，魏静，韩文霞，等．糖肾康中药复方含药血清对高糖环境下RMCs的增殖及TGF-β1/Smad2/3信号通路的影响[J]．中国中西医结合杂志，2015,35(1):88-92.
• [21]娄志杰，韩向莉，邵岩．糖尿病肾病中医病因病机及中医药内、外治疗研究新进展[J]．医学理论与实践，2017,30(6):808-810.
• [22]肖政华，杨辉，王和强，等．凌湘力教授之糖通饮方的相关研究[J]．中华中医药学刊，2019,37(8):1865-1868.
• [23]倪慧明，董哲毅，陈香美．黄芪主要活性成分对糖尿病肾病的疗效机制研究进展[J]．解放军医学杂志，2021,46(3):294-299.
• [24]王钦汶，戴新新，项想，等．丹酚酸和丹参酮干预糖尿病肾病的分子机制研究进展[J]．药学学报，2019,54(8):1356-1363.
• [25]马欢，高楠楠，陈俞如，等．糖通饮对2型糖尿病大鼠糖脂代谢的影响[J]．中成药，2021, 43(3):791-795.
• [26]潘艳伶，凌湘力．糖通饮对早期糖尿病肾病患者尿微量白蛋白排泄率、糖脂代谢的影响[J]．北方药学，2015,12(2):112-113.

文章评论(Comment)：