| Objective:In this study,the effect of Jujing Zhuyu granule on semen parameters of patients with asymptomatic asthenospermic infertility was observed in order to confirm the clinical effect of Jujing Zhuyu granule.At the same time,the basic components of the active substances in the compound prescription of Jujing Zhuyu granules were identified and the HPLC fingerprints of the prescriptions under different wavelengths were established in order to clarify the basic components and the whole fingerprint of Jujing Zhuyu granules,so as to make a preliminary exploration for the pharmacodynamic study of Jujing Zhuyu granules in the next step.Method:1According to the inclusion criteria,60 cases of asymptomatic asthenospermia infertility(4 cases)were treated with Jujing Zhuyu granule(1 dose/day,3 times/day,boiled water after meal,three times in the morning,middle and evening).The course of treatment was 13 weeks.The semen quality samples of patients before treatment,4 weeks,8 weeks and 13 weeks after treatment were collected strictly according to the requirements,and the relevant indexes in semen were analyzed by computer-aided sperm analysis system.according to the standards of the fourth edition of the World Health Organization Human semen examination and treatment Laboratory Manual,a self-control study was conducted before and after treatment.2 The study on the HPLC fingerprint of Jujing Zhuyu granule compound mainly includes the identification of the basic components of the active substances and the establishment of the fingerprint of the whole prescription.The single standard sample,the single particle sample,the negative control sample,the mixed standard sample 1,the mixed standard sample 2 and ten batches of particle samples of Jujingzhu granules were determined by HPLC-DAD technique.According to the determination results,the whole side dual-wavelength fingerprints(300nm and 210nm)of Jujingzhu granules were established,and the similarity evaluation system(2004A)of traditional Chinese medicine chromatographic fingerprint was used to evaluate them.Finally,the characteristic chromatographic peaks of HPLC fingerprints of Jujing Zhuyu granules were assigned unilaterally.Results:1 Study on the effect of Jujing Zhuyu granule on sperm motility in patients with asymptomatic asthenospermia:The semen related indexes of 56 patients with asymptomatic asthenospermia after 13 weeks of treatment were compared with those before treatment.The sperm density of 56 patients with asymptomatic asthenospermia after treatment(23.36±9.66×10~6/ml)was not significantly different from that before treatment(22.89±10.37×10~6/ml).The total sperm motility of patients after treatment(59.84±12.87%)was significantly different from that before treatment(46.82±10.34%)(P<0.05).Grade A sperm motility after treatment(37.10±12.07%)was significantly different from that before treatment(25.11±9.85%)(P<0.05).Grade B sperm motility after treatment(13.88±5.27%)was significantly different from that before treatment(11.33±7.23%)(P<0.05).Grade A+B sperm motility after treatment(50.97±11.92%)was significantly different from that before treatment(36.43±7.66%)(P<0.05).Among the 56 effective cases,the average age of the patients was 34.21±5.36years old,mainly between 26 and 35 years old,with the youngest being 23 years old and the maximum 45 years old.According to the standard of curative effect,8 cases were cured(14.29%),21 cases were markedly effective(37.50%),13cases were improved(23.21%),14 cases were ineffective(25.00%),and the total effective rate was 75.00%.2 Study on HPLC fingerprint of Jujing Zhuyu granules at different wavelengths:2.1 By comparing the chromatograms of single standard sample,single particle sample,negative control sample and mixed standard sample 1 and 2 of Jujing Zhuyu granule,it was found that the negative control sample decoction of Jujing Zhuyu granule had about 35chromatographic peaks at 300nm wavelength.After removing the chromatographic peak in the first 5 minutes,Danshen granule contributed 10 chromatographic peaks and Shengzhi Huangqi granule contributed 4 chromatographic peaks.Shengshu Dihuang granules contributed 2 chromatographic peaks,Chinese wolfberry granules contributed 4 chromatographic peaks,Jiuhuangjing granules contributed 1 chromatographic peak,Astragalus granules contributed7 chromatographic peaks,Chuanduan Dipsacus granules contributed 8chromatographic peaks,motherwort granules contributed 1 chromatographic peak,Polygonum multiflorum granules contributed 2 chromatographic peaks,while dodder granules,Taizishen granules and Jixueteng granules had no special chromatographic peaks.2.2 By comparing the chromatograms of the determination results of each single standard sample and each single particle sample,29 common peaks in the whole HPLC fingerprint of ten batches of Jujing Zhuyu granules under 300nm and 210nm wavelengths were identified,and 6 of them were identified.The chromatographic peaks of No.12,13,14,19,25 and29 are respectively Astragaloside,Rehmannia glutinosa,hyperoside(dodder)and stilbene glycoside(Polygonum multiflorum).The chromatographic peaks are Salvianolic acid B(Salvianolic acid B),Salvianolic acid B(Salvia miltiorrhiza),and No.29 saponins VI(Dipsacus).The other chromatographic peaks are made up of several drug particles.So it is difficult to identify and belong.Conclusions:Jujing Zhuyu Granules can effectively improve the patient’s sperm motility after 13 weeks of treatment.The chromatograms of Danshen granules,Chuanxuanduan granules,Shayuanzi granules,Shengzhi Huangqi granules and wolfberry granules in the negative control sample chromatograms of Jujingzhuyu granules at the wavelength of 300nm have the largest contribution;according to Fangyi of Jujingzhuyu granules According to the analysis,the 6 drugs with the most contributions in the chromatographic peak were Shengzhi Huangqi Granule and Wolfberry Seed Granule as the monarch medicine,Danshen Granule as the official medicine,Shayuanzi Granule as the adjuvant medicine,and Chuanxuandi Granule as the medicine,and the results Compatible with the drug compatibility of Jujing Zhuyu Granules,which further confirmed the effectiveness of the compatibility of Jujing Zhuyu Granules.Secondly,the chromatographic peak No.12 in the HPLC fingerprint of the ten batches of Jujing Assistant Granules at the wavelengths of 300 nm and 210 nm was identified as Verbascum isoflavone glucoside(Astragalus),peak No.13was verbascoside(Dihuang),and peak No.14 was gold.Hyperoside(Cuscuta chinensis)and stilbene glycoside(manufactured Polygonum multiflorum),No.19 chromatographic peak is Shayuanzi glycoside(Shayuanzi),No.25chromatographic peak is salvianolic acid B(danshen),No.29 chromatographic peak is Chuanxuantan Saponin VI(intermittent).It can be seen that the characteristics of the basic information of the Chinese medicinal compound medicinal substances in Jujing Zhuyu Granules are actually a comprehensive whole in the mixed state,which confirms the integration of the interaction and mutual interference of the ingredients of Jujing Zhuyu Granules.Function;and through the identified chromatographic peaks,it is known that it is compatible with Fang Yi,which further confirms the effectiveness of the combination of Jujing Zhuyu Granules. |
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