巴戟天经不同比例甘草炮制后对腺嘌呤致肾阳虚模型大鼠肾功能和HPG轴改善作用的比较

Effect of Morindae Officinalis Radix Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Improvement of Renal Function and HPG Axis of Kidney Yang Deficiency Model Rats Induced by Adenine

目的:通过比较巴戟天生品及不同比例甘草炮制品对腺嘌呤致肾阳虚模型大鼠肾功能和下丘脑-垂体-性腺轴(HPG轴)的改善作用,以及对大鼠组织中Wnt/β-连环蛋白(β-catenin)和转化生长因子-β_(1)( TGF-β_(1))/Smad信号通路蛋白表达的调节作用,分析不同比例甘草炮制(甘草与巴戟天质量比分别为0∶100、3∶100、6∶100、12∶100、24∶100,以下分别简称无甘草炮制品和100∶3、100∶6、100∶12、100∶24炮制品)对巴戟天补肾助阳功效的影响,阐释制巴戟天的炮制内涵。方法:巴戟天炮制品按2020年版《中华人民共和国药典》收载的制巴戟天方法进行制备。采用腺嘌呤灌胃给药复制肾阳虚大鼠模型,利用酶联免疫吸附测定法(ELISA)测定促卵泡激素(FSH)、黄体生成素(LH)、雌二醇(E2)、睾酮(T)水平,分光光度法测定尿素氮(BUN)和肌酐(SCr)水平,苏木素-伊红(HE)染色法评价组织病理形态变化,免疫组织化学法分析E-钙黏蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)、Wnt2b、β-catenin、Smad1和Smad4蛋白的表达。结果:100∶6、100∶12炮制品对体态的改善较好,100∶12炮制品降低血清中BUN、SCr、FSH和LH水平及调整E_(2)/T比值的作用最优,100∶6、100∶12炮制品调节E-cadherin、α-SMA、Wnt2b、β-catenin、Smad1和Smad4蛋白表达的作用较佳。结论:巴戟天经100∶6、100∶12比例甘草炮制后对腺嘌呤致肾阳虚模型大鼠肾功能和HPG轴有较好的改善作用,这与其能更好地调节大鼠肾脏和睾丸组织中Wnt/β-catenin通路及睾丸组织中TGF-β_(1)/Smads通路密切相关。

Objective:To explain the scientific connotation of Morindae Officinalis Radix(MOR)processed by Glycyrrhizae Radix et Rhizoma(Gly)by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly(GMOs)on the improvement of renal function and hypothalamic-pituitary-gonadal(HPG)axis,the protein expression of Wnt/β-catenin and transforming growth factor-β_(1)(TGF-β_(1))/Smad signal pathways in kidney Yang deficiency model rats induced by adenine.Method:GMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia.Rat model of kidney Yang deficiency was established by intragastrical administration of adenine,levels of follicle stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E_(2))and testosterone(T)were measured by enzymelinked immunosorbent assay(ELISA).Levels of urea nitrogen(BUN)and serum creatinine(SCr)were measured by spectrophotometry,hematoxylin-eosin(HE)staining was used to evaluate the pathological changes of kidney,testis and epididymis.Immunohistochemistry(IHC)was used to analyze the protein expression of E-cadherin,α-smooth muscle actin(α-SMA),Wnt2b,β-catenin,Smad1 and Smad4.Result:MOR processed with 100∶6 and 100∶12 proportions of Gly(short for GMO/100∶6 and GMO/100∶12)had the most obvious improvement on the body posture of kidney Yang deficiency model rats.GMO/100∶12 had the best effect on reducing the levels of BUN,SCr,FSH,LH and the ratio of E2/T.GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin,α-SMA,Wnt2b,β-catenin,Smad1 and Smad4.Conclusion:GMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine,which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β_(1)/Smads pathway in testicular tissue.