Rhubarb treatment of chronic renal failure study of the active ingredients
【Abstract】 Objective of medicinal rhubarb chronic renal failure study the chemical constituents. Methods chromatography and spectroscopy in the extraction and
isolation and identification of chemical constituents of rhubarb (-) epigallocatechin 3 o gallate (EGCG),
and through oral giving right nephrectomy-induced renal
failure rat model of treatment, observation of animals in each group blood urea nitrogen and serum creatinine levels. Results EGCG can reduce the renal failure rat model of blood urea nitrogen and serum creatinine levels and lower glomerular membrane injury index, and with a certain amount of drug dose-
effect relationship . Conclusion on chemical constituents of
medicinal rhubarb EGCG can kidney structure and physiological function near normal.
Key words chemical constituents of rhubarb renal failure
Study on the Chemical Constituent of Rhubarb in the Treatment of Chronic Renal Failure
Abstract: ObjectiveTo study the chemical constituent of rhubarb in the treatment of chronic renal failure. MethodsChromatographic and spectral technologies were used to extract, separate and elucidate the chemical constituent EGCG.Treat the kidney in the mouse model through giving orally, observes each group of animal urea nitrogen and the
serum myo anhydride content.ResultsThe level of urea nitrogen in the blood and serum myo anhydride decrease.EGCG
reducd the renal glomerulus's membrane damage
index.ConclusionThe chemical constituent EGCG of rhubarb can protect and repair the kidney organic function.
Key words: Medical rhubarb; Kidney failure; Chemical compostion
Chronic renal failure (Chronic Renal Failure, CRF) is a variety of renal diseases, the ultimate outcome of continued progress, but to stop its progress is still the most concern
clinicians and patients one of the issues. The treatment of CRF include renal replacement therapy and non-replacement
therapy. At present can only be used to extend the life of dialysis treatment rather than to fundamentally improve the
renal function; renal transplantation there is higher cost for the source of tension and there is rejection and complications and other issues. Rhubarb and its compound preparations have been widely used in clinical treatment of CRF . It inhibited the development of the precise effect of CRF [2,3]. According to literature reports [4 ~ 6], rhubarb effective in
inhibiting CRF mainly composed of () table galloyl catechins
3 o gallate () epigallocatechin 3 o
gallate) (EGCG ). This focus on the extraction and separation of EGCG, structure identification and suppression CRF preliminary pharmacological tests to study for the treatment of their chronic renal failure provide the basis for drug research in Chinese medicine.
An apparatus and materials
XRC 1 micro-melting point-type detector (temperature
uncorrected); IR 440 IR spectrometer; Varian INOVA600
nuclear magnetic resonance instrument (TMS as internal standard); thin layer chromatography silica gel and column chromatography silica gel (Qingdao Marine Chemical Factory); Sephadex LH 20 gel (Pharmacia company); other reagents were
of analytical grade. EGCG standard were purchased from SIGMA, Inc.; rhubarb purchased from Hubei Lichuan medicine company, from Hubei College of Professor Chen Keli identified as Rheum officinale Baill. Pure Wistar rats, normal level, provided by
the Hubei Traditional Chinese Medicine. EGCG, according to the methods described in the text after the extract preparation.
2 Methods and Results
2.1 EGCG Extraction, Isolation and identification of
2.1.1 Extraction and Separation of dried rhubarb meal
with 20 times the amount of 95% ethanol at room temperature percolation, vacuum extraction solvent was waving to make extracts, to four times the amount of water dispersion so-
miscible, and then ethyl acetate extract equivalent five times, vacuum solvent recovery to be cream. Recycled water-
phase dispersion, the chloroform extraction five times, the water layer after repeated silica gel chromatography, gradient elution of methanol with chloroform and Sephadex LH 20
column chromatography, gradient elution of methanol with water
be compound EGCG.
2.1.2 Structure Identification of compounds EGCG: light yellow powder, mp218 ?. Soluble in water, methanol and other polar solvents. FeCl3 responses of blue, with the reference substance EGCG were thin, toluene: ethyl acetate: formic acid (5:4:1) started the same Rf value. 1H NMR (Acetone d6)
δ: 2.95 (1H, d, C 4e H), 3.02 (1H, d, C 4a H),
5.10 (1H, d, C 2 H), 5.49 (1H, t,, C 3 H), 6.01
(1H, s, C 8 H), 6.03 (1H, s, C 6 H), 6.60 (2H, s,
C 2 ' H, C 6 ' H), 7.01 (2H, s, C 2'' H, C
6'' H). 13C NMR (Acetone d6) δ: 25.97 (C4), 68.66
(C3), 77.39 (C2), 95 (C8), 95.8 (C6), 98.2 (C10), 106 (C2 ', C8'), 109.2 (C2'', C6''), 121.1 (C1''), 130 (C1 '), 132.5
(C4'), 138.1 (C4''), 145.1 (C3'', C5''), 145.5 (C3 ', C5'), 156 (C5, C7), 157 (C9), 165.4 (C7'').
The above data and reported in the literature [7,8] of the spectral data, and therefore identified the powder as EGCG.
2.2 EGCG pharmacological experiments to remove the
healthy 20 male rats, intraperitoneal injection of 2.5% pentobarbital anesthesia. Line along the ventral midline abdominal incision, making the whole right kidney removed, and then cut out most of the left kidney , residual left kidney
treated by bleeding and suture the wound, sub-cage rearing.
Cut off the tail at intervals of 10 d after blood sampling determination of urea nitrogen, blood urea nitrogen to reach 62 mg, that he had kidney failure. Draw lots to decide grouping, the control group each watering 10 ml / d; treatment group ? each fed EGCG10 mg / d; treatment group ? each fed
EGCG 20 mg / d. Reposted elsewhere in the paper for free download http://
2.2.1 EGCG effect on blood urea nitrogen in Table 1. Table 1 EGCG on the impact of renal failure in rat model of blood urea nitrogen (abbreviated)
2.2.2 EGCG effects on serum creatinine in Table 2. Table 2 EGCG on the renal failure rat model the impact of serum creatinine (omitted)
As can be seen from Table 1, with the extension of
experimental time, blood urea nitrogen level of the control group gradually increased; and two treatment groups were gradually reduced to 20 d, when that is a significant difference.
As can be seen from Table 2, with the prolonged
experiment, three groups of serum creatinine levels were elevated, but compared with the reference substance, to 20 d when the two treatment groups of serum creatinine level has been a noticeable difference, especially in the late control rats had a marked uremic symptoms, the treatment group maintained a lower level.
2.2.3 Histopathological observations in Table 3. Table 3 Histopathological observation (omitted)
Normal rat mesangial cells are located in between the basement membrane is not coated capillary, attached to the vessel wall, its physiological function is to play supporting role to the capillaries, the glomerular capillary wall in the filtration material left over from the There scavenging effect. Renal failure in rat mesangial cells cytoplasm
swelling, filtration rate is low, through the capillary wall material left behind and more need to remove the material in mesangial cells are also more cells removed more than large particles, thus causing mesangial cells damage are also great,
matrix showing hyperplasia and fibrosis; while the treatment group was left behind by the capillary filtration and the material less mesangial cells and therefore removed the material particles are small and few, and also because removal of light load, causing the Department of mesangial cell matrix proliferation and fibrosis are also light.
Nitrogen metabolism disorder is a prominent manifestation of CRF patients with a large number of experimental and clinical studies have shown that rhubarb could improve
nitrogen metabolism . Preliminary experiments done more than EGCG on serum creatinine and urea nitrogen effects of pharmacological experiments confirmed that reducing serum creatinine and urea nitrogen levels.
Glomerular mesangial cell proliferation and extracellular matrix is an increase in glomerular sclerosis  in the process of the main performance of mesangial cells are vascular pericytes, is active in response to glomerular intrinsic cells in the inflammatory process is not However,
passive victims, but also a direct participant, through changes in its structure and metabolism of a direct impact on the inflammatory process, these experiments confirmed that, EGCG for the protection and restoration of the role of kidney
tissue, reducing blood urea nitrogen, lower serum creatinine, so that kidney filtration and excretion of metabolites of near-normal function, reducing waste in the body retention; the retention of waste in the body in turn reduce the poisoning and damaged kidney cells to reduce, so that the organizational structure and physiological function of kidney was near normal.
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