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Ramulus Mori (Sangzhi) alkaloids (SZ-A) are a group of polyhydroxy alkaloids extracted and isolated from the traditional Chinese medicine mulberry twig, which is mainly used for the treatment of type 2 diabetes mellitus (T2DM). In addition to acting as a glycosidase inhibitor in the small intestine after oral administration, SZ-A can also be absorbed into blood and widely distributed to target organs related to diabetes, exerting multiple pharmacological effects. It is important to elucidate the possible pharmacokinetic influences of SZ-A for its clinical rational applications, such as drug interactions, the effects of food and alcohol on the absorption of SZ-A. However, studies in this area are limited. Therefore, the pharmacokinetic interactions between orally administrated SZ-A (50 mg·kg-1) and metformin hydrochloride (Met, 200 mg·kg-1) in Sprague-Dawley (SD) rats were examined. Then, the effect of food (standard feed) on the pharmacokinetics of SZ-A was investigated using fasting administration of SZ-A (50 mg·kg-1) in rats as a control. Finally, we investigated the pharmacokinetic characteristics of SZ-A (50 mg·kg-1) in different concentrations alcohol solutions using aqueous solution of SZ-A administered to rats as a control to evaluate the effect of alcohol on the bioabsorption of SZ-A. The results showed no significant pharmacokinetic interactions between SZ-A and Met after combination treatment. The standard feed had little effect on the pharmacokinetic profile of SZ-A. Alcohol retarded the absorption of SZ-A, resulting in a significant decrease in the Cmax of SZ-A. The decrease was greater at higher alcohol concentrations; however, no significant difference was observed in the AUC0-t. These results support the clinical rational applications of SZ-A. All animal protocols were approved by the Ethics Committee of Kangtai Medical Laboratory Service Hebei Co., Ltd. (Hebei, China) (No. MDL2022-01-17-1).
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OBJECTIVE： To study on the effects of prophylactic administration of Ramulus mori polysaccharides （RMP） on inflammatory response of renal ischemia reperfusion injury （RIRI） model mice and to explore its possible mechanism. METHODS： Totally 60 C57BL/6 mice were randomly divided to sham operation group， model group， atorvastatin group （positive control， 15 mg/kg）， RMP low-dose， medium-dose and high-dose groups （300， 600， 1 200 mg/kg）. Except for sham operation group， RIRI model was induced in other 5 groups. 24 h before surgery， they were given relevant medicine intragastrically， once a day， for consecutive one week. 24 h after reperfusion， the mice were sacrificed. The serum levels of Scr and BUN were detected. The morphological changes of renal tissue were observed under optical microscope. The serum levels of IL-1β， IL-6， IL-10 and TNF-α were determined by ELISA. Western blot assay was used to determine the protein expressions of Toll-like receptor 4 （TLR4）， p38 mitogen-activation protein kinase （p38MAPK） and p-p38MAPK. RESULTS： Compared with sham operation group， the serum levels of Scr and BUN were significantly elevated in model group （P＜0.01）. RIRI led to typical inflammatory response of renal tissue， widespread renal tubular epithelial cell degeneration and necrosis， and inflammatory cells infiltration. Serum levels of IL-1β， IL-6， IL-10 and TNF-α were increased significantly （P＜0.01）. The protein expressions of TLR4， p38MAPK and p-p38MAPK were increased significantly in renal cortex （P＜0.01）. Compared with model group， serum levels of Scr and BUN were decreased significantly in administration groups （P＜0.05 or P＜0.01）. The pathological damage of renal tissue was improved in varying degrees， especially in the RMP medium-dose and high-dose groups. Serum levels of IL-1β and IL-6 were decreased significantly in administration groups （P＜0.05 or P＜0.01）. Serum levels of IL-10 were further increased in atorvastatin group and RMP high-dose group （P＜0.01）， and serum level of TNF-α was decreased significantly in atorvastatin group and RMP medium-dose and high-dose groups （P＜0.05 or P＜0.01）. The protein expressions of TLR4 and p-p38MAPK in renal cortex were decreased significantly in administration groups （P＜0.05 or P＜0.01）. CONCLUSIONS： RMP prophylactic administration can improve RIRI of mice， the mechanism of which may be associated with relieving the inflammatory response through inhibition of TLR4/p38MAPK signaling pathway.
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Objective To extract the volatile oil of Ramulus Mori and analyze the chemical composition of the volatile oil. Methods The volatile oil of Ramulus Mori was extracted by simultaneous distillation extraction(SDE)and direct distillation extraction(DDE)method, and the chemical composition was identified by GC-MS. Results Nineteen compounds were identified in the volatile oil extracted by SDE, which accounted for 94.49% of the total amount of detection. There were alkyl, acids, aldehydes, ketones, esters, alcohols and other substances in the volatile oil. And 16 compounds were determined in the volatile oil extracted by DDE, accounting for 97.12% of the total amount detected. There were furan, carboxylic acids, esters, aldehydes, alcohols, phenols, ketones, alkene and acyl halide. The chemical compositions of the volatile oil extracted by SDE were more than those extracted by DDE. Conclusion The chemical constituents of volatile oil of Ramulus Mori can be more fully identified by the combination of the two extraction methods.
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Objective To extract the volatile oil of Ramulus Mori and analyze the chemical composition of the volatile oil. Methods The volatile oil of Ramulus Mori was extracted by simultaneous distillation extraction(SDE)and direct distillation extrac?tion(DDE)method,and the chemical composition was identified by GC-MS. Results Nineteen compounds were identified in the vol?atile oil extracted by SDE,which accounted for 94.49%of the total amount of detection. There were alkyl,acids,aldehydes,ketones, esters,alcohols and other substances in the volatile oil. And 16 compounds were determined in the volatile oil extracted by DDE,ac?counting for 97.12 % of the total amount detected. There were furan,carboxylic acids,esters,aldehydes,alcohols,phenols,ke?tones,alkene and acyl halide. The chemical compositions of the volatile oil extracted by SDE were more than those extracted by DDE. Conclusion The chemical constituents of volatile oil of Ramulus Mori can be more fully identified by the combination of the two ex?traction methods.
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Objective To establish a thin-layer chrometograrrgy(TLC) method for the identification of quercetin and chlorogenic acid in Ramulus mori, and to screen their antioxidant activity. Methods The quercetin and chlorogenic acid were extracted by ultrasonic method with methanol as a solvent. The effect of different developed system, reagent, temperature, view methods and different silica gel plate on the TLC of quercetin and chlorogenic acid in Ramulus mori were tested to select the best TLC conditions. The antioxidant activity of quercetin and chlorogenic acid was screened with DPPH as a reagent. Results The ethyl acetate:water:formic acid:toluene(17:2:2:0.8) was used as a developing system and 1% AlCl3 as a chromogenic reagent. Quercetin and chlorogenic acid in Ramulus mori were identified under 366 nm, with blue and blue-green spots on silica gel G plate, and yellowish spots under purple background by the test of TLC-bioautography. Both were proven to have antioxidant activity. Conclusion The method is simple, accurate and reliable, and can be used for quality control of Ramulus mori.
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Objective To establish the methods for identification and quantification of γ-aminobutyric acid (GABA) in Ramulus Mori by TLC and HPLC. Methods GABA was extracted by ultrasonic method with ethanol as solvent. The sample was applied on an efficient silica gel G plate with n-butanol-acetic acid-water (4∶2. 2∶1) as the developing system, and ninhydrin was used as chromogenie reagent. The content of γ-GABA was determined by HPLC. The separation was carried out on a ODS (4. 6 mm×250 mm, 5 μm) column with phosphate buffered solution (pH=6. 8)-methanol as a mobile phase with gradient elution, flow rate was 1. 0 ml/min, the detection wavelength was 335 nm. Results The TLC spots of GABA in Ramulus Mori were clear with good separation. The assay of GABA was linear in the range of 1. 006-84. 504 μg/ml and correlation coefficient was 0. 9994. Conclusion The methods are simple, accurate and reproducible which are valuable to identify Ramulus Mori.
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Aim To investigate the influence of Ramu-lus mori polysaccharides ( RMP) on blood glucose and anti-oxidative effect in streptozotocin ( STZ )-induced diabetic mice .Methods Diabetic mice were induced by intraperitoneal injection with 120 mg? kg -1 STZ and were randomly divided into the following 5 groups with 20 animals per group: model group , valsartan group ( 20 mg? kg -1 ) , low-, medium-, high-dose (0.3,0.6,1.2 g? kg -1 ) of RMP groups.Other 20 normal mice were treated as normal control group .The mice were administered orally for 90 d.On 45 d of ad-ministration , the 24 h urine was collected through met-abolic cages for urine protein detection .Pathological changes of kidney tissues were observed through HE staining .The serum levels of urea nitrogen ( BUN) and creatinine ( Cr ) were detected by automatic biochemical analyzer; and the manganese superoxide dismutase (Mn-SOD), catalase(CAT), malonaldehyde(MDA) and mitochondrial respiratory chain complex Ⅰ,Ⅲac-tivity of kidney tissues were also determined .ELISA method was used to detect ROS content in renal cortex . The SIRT1 , FOXO1 and NF-κB protein expressions were analyzed by Western blot .Results Compared with model group, the FBG, microalbuminuria, BUN and Cr were decreased by RMP medication ( P <0.05).The activities of Mn-SOD, CAT and mitochon-drial respiratory chain complex Ⅰ,Ⅲ in RMP groups were enhanced , while MDA and ROS levels were re-duced. Moreover, the expressions of SIRT1 and FOXO1 were up-regulated by RMP , the expression of NF-κB was down-regulated ( P<0.05) .Conclusion RMP exerts renal protective effect through up-regula-ting the expressions of SIRT1 and FOXO1 in renal cor-tex , which may relate to the improvement of anti-oxida-tive capability .
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Objective To establish the methods for identification and quantification ofγ-aminobutyric acid(GABA)in Ramu?lus Mori by TLC and HPLC. Methods GABA was extracted by ultrasonic method with ethanol as solvent. The sample was applied on an efficient silica gel G plate with n-butanol-acetic acid-water(4∶2.2∶1)as the developing system,and ninhydrin was used as chromog?enie reagent. The content ofγ-GABA was determined by HPLC. The separation was carried out on a ODS(4.6 mm × 250 mm,5μm) column with phosphate buffered solution(pH=6.8)-methanol as a mobile phase with gradient elution,flow rate was 1.0 ml/min,the de?tection wavelength was 335 nm. Results The TLC spots of GABA in Ramulus Mori were clear with good separation. The assay of GA?BA was linear in the range of 1.006-84.504μg/ml and correlation coefficient was 0.9994. Conclusion The methods are simple,ac?curate and reproducible which are valuable to identify Ramulus Mori.
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Objective To establish a thin-layer chrometograrrgy(TLC)method for the identification of quercetin and chloro?genic acid in Ramulus mori,and to screen their antioxidant activity. Methods The quercetin and chlorogenic acid were extracted by ultrasonic method with methanol as a solvent. The effect of different developed system,reagent,temperature,view methods and differ?ent silica gel plate on the TLC of quercetin and chlorogenic acid in Ramulus mori were tested to select the best TLC conditions. The antioxidant activity of quercetin and chlorogenic acid was screened with DPPH as a reagent. Results The ethyl acetate∶water∶formic acid∶toluene(17∶2∶2∶0.8)was used as a developing system and 1%AlCl3 as a chromogenic reagent. Quercetin and chlorogenic acid in Ramulus mori were identified under 366 nm,with blue and blue-green spots on silica gel G plate,and yellowish spots under purple background by the test of TLC-bioautography. Both were proven to have antioxidant activity. Conclusion The method is simple,accu?rate and reliable,and can be used for quality control of Ramulus mori.
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Objective To optimize the ultrasonic extraction technology of flavonoids from Ramulus Mori. Methods Ultrasonic extraction technology was optimized by orthogonal test,and the content of total flavonoids was determined by spectrophotometry. Results The optimal condition was:extracting for 3 times and 20 min for each time,with 8 fold of 80% ethanol. Conclusion The ultrasonic extraction technology was simple,rapid and high efficient.
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BACKGROUND: The exact role of aerobic bacteria in the pathogenesis of acne remains controversial but several reports showed that these microorganisms were important in the development of acne lesions and affected the severity of acne by being involved in the inflammatory process. There were several attempts to improve and prevent the acne lesions by using soap or cosmetics containing anti-microbial agents. OBJECTIVE: The purpose of this study was to compare the sebum secretion rates and the number of aerobic bacteria in acne patients after 4 weeks trial of RT cream containing 0.1% tea-tree oil and 0.01% Ramulus mori extract. METHODS: We studied 10 normal controls and 20 acne patients. Sebum secretion was measured by Sebumeter. Clinical efficacy was evaluated by comparing numbers of non-inflammatory and inflammatory lesions. Aerobic bacteria was also cultured from control group and acne patients group before and after RT cream use. RESULTS: Sebum secretion rates in acne patients were higher than those in control, but not statistically significant. Total number of aerobic bacteria in acne patients was higher than that in control (p<0.05). The aerobic bacteria consisted of coagulase negative Staphylococcus, Streptococcus sp., S. aureus in both acne patients and control. After 4 weeks, the number of inflammatory lesions had decreased (p<0.05%). In addition, the number of aerobic bacteria showed the tendency of decrease after RT cream use. CONCLUSION: RT cream consisting of 0.1% tea tree oil and 0.01% Ramulus mori extract was found to be effective and safe for acne patients with especially inflammatory lesions.
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Humans , Acne Vulgaris , Anti-Infective Agents , Bacteria, Aerobic , Coagulase , Sebum , Skin , Soaps , Staphylococcus , Streptococcus , Tea Tree Oil , TeaABSTRACT
AIM: To study the isolation,purification and characterization of Ramulus Mori polysaccharide. METHODS: Ramulus Mori was extracted by boiling water. The raw extract was precipitated fractionally by alcohol deproteinized,passing through DEAE ion exchange cellulose ( DEAE-52) and SephadexG-100,obtained RMPS1 and RMPS2. The composition and characterization of Ramulus Mori polysaccharide were researched by TLC、IR、 GC、HPLC and smith degradation. RESULTS: The molecular weight of RMPS1 and RMPS2 were 5. 8 ? 105 and 6. 5 ? 105; RMPS1was made up of rhammose、arabinose、glucose and galactose with the molarity rate of 1. 08 ∶ 1 ∶ 1. 40 ∶ 1. 57; RMPS2 of rhammose、glucose and galactose with the molarity rate of 11. 38 ∶ 1 ∶ 1. 35. Smith degradation showed that the main linkage form in RMPS1 and RMPS2 was 1→2 and 1→4 glycosidic linkages,But some 1→3 glycosidic linkages also existed in the molecules; infrared spectrum showed that both had the polysaccharide characteristic absorption peaks. CONCLUSION: The structures of RMPS1 and RMPS2 are first determined from Ramulus Mori.