Research On The Efficacy Of Crude And Related Processed Rhizoma Pinelliae Based On Material Basis And Biological Activity

Pinelliae rhizoma, belonging to the family Araceae, is one of the Traditional Chinese Medicinal herbs. It was first recorded in "Shen Nong’s Herbal Classic" and the raw pinelliae rhizoma was classified into the third class according to the book because of its negligible toxicity. The pinelliae rhizoma’s property is warm and possesses various medical activities such as dislodging dampness and resolving phlegm, calming stomach and preventing emesis, dissolving lumps and resolving masses. The oral administration of raw pinelliae rhizoma is prohibited because of its raw material could benumb the tongue and stimulate throat then lead to numbness, swelling, emesis even asphyxia. Therefore, the pinelliae rhizoma is always used after preparation. There are four pinelliae rhizoma processed products viz. pinelliae rhizoma praeparatum cum alumine, pinelliae rhizoma praeparatum cum zingibere et alumine, pinelliae rhizoma praeparatum and Jing Pinelliae. The former three have been recorded in the "Chinese Pharmacopoeia"(2010, Volume I). There are various processed products of Pinelliae rhizoma and their chemical components and pharmacological effects are quite different according to the processed methods. The above problem puzzles the quality control o pinelliae rhizoma and it’s processed product for a long time. To date, there is no effective method to solve this problem.This paper systematically investigated the medical effects’changes of raw pinelliae rhizoma and its four processed products through three aspects, viz. chemical components, pharmacodynamics and bioactivity, illuminated the relativity of chemical components and pharmacodynamics, bioactivity before and after preparation, revealed how the change of effective ingredients basis before and after preparation of pinelliae rhizoma influenced the medical effects and bioactivity, indicated the anti-tussive effect of total organic acids and the the anti-inflammatory effect of total alkaloids could be used as the reference quality compounds of aw pinelliae rhizoma and its four processed products. We also established the evaluation methods of inherent quality of raw pinelliae rhizoma and its four processed products based on the changes of HPLC character fingerprints, pharmacological activity and bioactivity, and provided a new idea for improving the quality standard and establishing the overall and diversification quality control mode of traditional Chinese medicine quality control mode. Here are the main results:1、Physical and chemical analysis of Pinellia rhizoma and its processed products1.1This article illuminated the change of effective ingredients basis before and after preparation of pinelliae rhizoma by qualitative and quantitative analysis of chemical compositions. We measured the contents of total organic acids and total alkaloids of raw pinelliae rhizoma and its processed products by potentiometric titration and acid dye colorimetry. The total organic acids contents of all processed products increased at different level after the preparation. The total organic acids contents of Jing Pinelliae, Rhizoma Pinelliae Praeparatum and Rhizoma Pinelliae Praeparatum Cum Alumine increased by25%,20%,15%compared to pinelliae rhizoma. Except the content of total alkaloids of Rhizoma Pinelliae Praeparatum Cum Zingibere Et Alumine increased by20%, Jing Pinelliae, Rhizoma Pinelliae Praeparatum and Rhizoma Pinelliae Praeparatum Cum Alumine decreased by12%,38%and50%compared to pinelliae rhizoma.1.2It was the first time to establish the characteristic fingerprints of different parts (water,75%alcohol and95%alcohol) in pinelliae rhizoma and its processed products by HPLC. We also identified8characteristic peaks, viz. inosine, guanosine, adenosine, succinic acid, ephedrine hydrochloride, liquiritin, ammonium glycyrrhetate,6-gingerol.6-gingerol was the unique characteristic peak of pinelliae rhizoma praeparatum cum zingibere et alumina while liquiritin and ammonium glycyrrhetate are unique characteristic peaks of pinelliae rhizoma praeparatum and Jing pinelliae respectively. Furthermore, compared with pinelliae rhizoma, pinelliae rhizoma praeparatum cum alumine and pinelliae rhizoma praeparatum cum zingibere et alumine had an new peak at18.3min while pinelliae rhizoma praeparatum and Jing pinelliae had an new peak at45.4min without inosine.2、Pharmacological effects of Pinellia rhizoma and its processed products2.1We studied the pharmacological effects of the chemical compounds of Pinellia rhizoma and its processed products. The results indicated that total organic (?)loids and total phenolic acids are the three active ingredients of Pinellia rhizoma and its processed products, showed anti-tussive, anti-inflammatory and antioxidant activities, respectively.2.2The study on total organic acids content and its anti-tussive activity of pinelliae rhizoma and its processed products indicated that Jing pinelliae and pinelliae rhizoma praeparatum behaved the highest total organic acids content and showed the highest anti-tussive activity; pinelliae rhizoma praeparatum cum alumine and pinelliae rhizoma praeparatum cum zingibere et alumine had higher total organic acids content and showed higher anti-tussive activity. Pinelliae rhizoma had the lowest total organic acids content and showed the lowest anti-tussive activity. In the clinical use of traditional Chinese medicine, the application of pinelliae rhizoma praeparatum and Jing pinelliae emphasis on anti-tussive because of concocted process by adding alicorice. The application of Rhizoma Pinelliae Praeparatum Cum Alumine emphasis on expectprant and reduce lung air, its anti-tussive activity is weaker than pinelliae rhizoma praeparatum and Jing pinelliae. Pinelliae rhizoma praeparatum cum zingibere et alumina emphasis on calming stomach and preventing emesis because of adding ginger. Pinelliae rhizoma main topical application, not orally. Our research indicated that there existed a close relationship between total organic acids and anti-tussive activity. The anti-tussive activity could be higher when the total organic acids content is higher and the total organic acids’s anti-tussive activity was consistent with the clinical application. Therefore, it’s reasonable for "Chinese Pharmacopoeia"(2010, Volume I) to apply total organic acids content as quality standard for pinelliae Yiizoma and its processed products.2.3There is a close relationship between active ingredients and pharmacological activities of traditional Chinese medicine. The former reflect the later and the later is the expression of the former. Therefore, we performed a correlation analysis of active ingredients and pharmacological activities. The result indicated that the relativity existing between total organic acids and anti-tussive activity, total alkaloids and anti-inflammation activity, total phenols and antioxidant activity. Futhermore, the activities were enhanced while the dosage increased.2.4It was the first time to survey the "chromatography-effect" relationship of total organic acids’anti-tussive activity of pinelliae rhizoma and its processed products. The results showed that the chemical compositions represented by peak 0.4, NO.6(succinic acid) and NO.12had strong relevancy with anti-tussive activity and contributed the most to this activity. What’s more, the chemical compositions represented by peak NO.4with large peak area and high relevancy could used as a new reference compound to assess the quality of pinelliae rhizoma and its processed products and deserved futher research about its structure.3、Bioactivity of of pinelliae rhizoma and its processed products3.1It was the first time to establish the determination methods of anti-tussive titer by total organic acids and anti-inflammation titer by total alkaloids for pinelliae rhizoma and its processed products. We established the method for determination of biological potency of pinelliae rhizoma and its processed products and obtained their titer’s value, starting with the pharmacological effects, basing on mouse strong aqua ammoniac induced cough model and foot swelling affecting inflammatory mediator PGE2model. The results demonstrated that the anti-tussive titers of pinelliae rhizoma and its processed products increased after prepared. Jing pinelliae showed the highest anti-tussive titer with2105.59U·g-1, followed by pinelliae rhizoma praeparatum, pinelliae rhizoma praeparatum cum alumine and pinelliae rhizoma praeparatum cum zingibere et alumine. Pinelliae rhizoma existed the lowest anti-tussive titer, only33.3%compared to Jing pinelliae. However, the anti-inflammation titers of pinelliae rhizoma and its processed products decreased after prepared. Pinelliae rhizoma showed the highest anti-tussive titer with8.49U·g-1followed by pinelliae rhizoma praeparatum cum zingibere et alumine, Jing pinelliae and pinelliae rhizoma praeparatum. Pinelliae rhizoma praeparatum cum alumine showed the lowest anti-tussive titer, only25%compared to Pinelliae rhizoma.3.2The study on biological potency and pharmacological activity of pinelliae rhizoma and its processed products demonstrated that the total organic acids of Jing pinelliae and pinelliae rhizoma praeparatum had the biggest anti-tussive titer value, showed the highest anti-tussive activity; while pinelliae rhizoma praeparatum cum alumine and pinelliae rhizoma praeparatum cum zingibere et alumine had bigger anti-tussive titer value, showed higher anti-tussive activity. Pinelliae rhizoma had the smallest anti-tussive titer value and showed the lowest anti-tussive activity. Thus, we could concluded that there existed a close relationship between biological potency value and pharmacological activity. The anti-tussive activity could be higher when the anti-tussive titer value is bigger and the determination result of the total organic acids’s anti-tussive titer was consistent with the clinical application. Total organic acids’s anti-tussive assay can used to evaluate the quality of pinelliae rhizoma and its processed products which better serve their clinical application. The result has significant meaning for ensuring the safety and efficacy of traditional Chinese medicine.3.3Biological potency reflects the information of pharmacological activity and the active ingredients are the basis of pharmacological activity. Thus, in order to illustrate the relationships between total organic acids and anti-tussive titer, total alkaloids and anti-inflammatory titer, we performed a correlation analysis between biological potency and active ingredients. The results indicated that the anti-tussive effect and anti-tussive titer increased when the content of total organic acids is higher. The content of total organic acids showed the same trend of anti-tussive titer, expressing a significant positive correlation between them. The content of total alkaloids also showed the same trend of anti-inflammatory titer.In conclusion, the study on the correlation analysis between the active ingredients with pharmacological activities and bioactivities of Pinellia ternata and its processed products indicated that the total organic acids of Pinellia ternata and its processed products exhibited antitussive activity and the total alkaloids had anti-inflammatory activity which can be use as the quality evaluation standard of them. We also found that the combination of HPLC character fingerprints, pharmacological activity and bioactivity effectively evaluate the quality of pinelliae rhizoma and its processed products and serve as a new quality standard method. The method could improve the quality standards of pinelliae rhizoma and its processed products and provide experimental basis to build a new mode for traditional Chinese medicine.