Effects Of Different Extracted Parts Of Cistanche Deserticola On Hypoglycemic Effects Of Db / Db Black Mice

Abstract: 

Objective: To explore the effects and mechanisms of different extracted parts of Cistanche deserticola and Tartschula deserticola on lowering blood sugar in db / db black mice. 

Method: The total polysaccharides, oligosaccharides, and glycosides of Cistanche deserticola and Tartschula deserticola were enriched, and their contents were compared and analyzed. db/db black mice were selected as experimental animals. After 28 days of administration, the body weight, food intake, and water consumption of the black mice were recorded, and blood glucose was detected. ELISA detected serum insulin (INS), glycosylated hemoglobin (HbA1c), and liver triglyceride. ester (TG), total cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), malondialdehyde (MDA), superoxide dismutase (SOD), plasma, and urine The level of uric acid (UA), creatinine (CR), and urine micro protein (MALB) levels in liquid; the HE staining method is prepared for the pathological slices of pancreatic and kidney tissue; IHC measures the level of protein expression of the kidney TYPE -IV Collagen.

 Results: The content of the polysaccharide part of Cistanche deserticola decreased after steaming with wine. The content of betaine in the oligosaccharide part of Cistanche deserticola increased after steaming with wine. The content of total glycosides echinaceaside slightly decreased after steaming with wine. The content of verbascoside decreased. The content of heterogeneous leaves Cimicifugarin content increased. Compared with the model group, different extracted parts of Cistanche deserticola and Tartscholaris can reduce body weight, fasting blood glucose (FBG), oral glucose tolerance (OGTT), HbA1c, insulin resistance index (HOMA-IR), islet β-cell secretion index (HOMA- β), MDA, LDL-C, UA, Cr and mALB, INS and HDL-C have an upward trend (P<0.05, P<0.01), and can improve the pathological damage of pancreas and kidney and ty ｐｅＩＶ Expression of collagen. 

Conclusion The total glycosides of Cistanche deserticola and Tartschalis deserticola have better hypoglycemic effects, and the polysaccharides and oligosaccharides of Tartschalne deserticola also have certain hypoglycemic effects.

Keywords: Cistanche deserticola; total polysaccharides; total oligosaccharides; total glycosides; db / db black mice; hypoglycemia

ORGANIC CISTANCHE EXTRACT WITH 25% ECHINACOSIDE AND 9% ACTEOSIDE

Cistanche deserticola is the dry fleshy stem with scaly leaves of Cistanche deserticola or Cistanche deserticola. Dig the seedlings in spring when they are newly unearthed or before the freezing soil in autumn, remove the stem tips, cut them into sections, and dry them in the sun [1]. Cistanche deserticola has the effects of nourishing kidney yang, replenishing essence and blood, and moisturizing the intestines and laxatives. It is clinically used for insufficient kidney yang, deficiency of essence and blood, impotence and infertility, soreness and weakness of the waist and knees, weakness of muscles and bones, intestinal dryness, and constipation. The pharmacological effects of Cistanche deserticola include anti-aging, antioxidant, anti-dementia, anti-fatigue, and laxative effects [2-3]

. The main components of Cistanche deserticola include phenylethanoid glycosides, iridoid glycosides, polysaccharides, etc., among which phenylethanoid glycosides Alcohol glycosides, and polysaccharides are the main medicinal substances [4].

Modern pharmacological research shows that Cistanche deserticola polysaccharide has many pharmacological effects such as regulating immune activity, anti-aging, improving learning and memory ability, protecting nerves, anti-liver damage, anti-virus, anti-tumor, affecting intestinal flora [5]. Cistanche deserticola total glycosides have the functions of nourishing the liver and kidney, replenishing essence and blood, antioxidant, anti-aging, immune enhancement, and neuroprotection [6]. "Chinese Pharmacopoeia" 2020 Edition

(Part 1) contains two processed varieties of Cistanche deserticola tablets and Liquor deserticola. Preliminary research by the research team found that the raw product of Cistanche deserticola tends to moisturize the intestines and relieve constipation, while Liquor deserticola has significantly enhanced kidney-tonifying and yang-enhancing effects. The total glycosides and total polysaccharides of Cistanche deserticola are the effective parts for replenishing kidney yang, and the oligosaccharides of Cistanche deserticola are the effective parts for moisturizing the intestines and laxative [7]. During the wine-steaming process of Cistanche deserticola, the phenylethanoid glycoside components changed significantly, the content of verbascoside decreased, while the content of its isomer verbascoside increased significantly [8]. Type 2 diabetes is a metabolic disease caused by insulin resistance, accompanied by structural and functional damage to pancreatic β-cells, which in turn leads to insufficient insulin secretion and is mainly characterized by elevated blood sugar [9]. Traditional Chinese medicine believes that diabetes is often related to kidney essence, which stores essence. Deficiency of essence can induce diabetes and even lead to diabetic nephropathy. Symptoms of type 2 diabetes mostly include excessive urination, fatigue, thirst and thirst, and hunger after eating too much, which are similar to those of kidney yang deficiency. Therefore, the clinical treatment of diabetes in traditional Chinese medicine focuses on tonifying the kidney and replenishing the kidney yang [10-11].

  According to reports in the literature, the phenylethanol glycosides in Cistanche deserticola, such as echinaceaside and verbascoside, can not only inhibit the increase in postprandial blood sugar levels but also improve the glucose tolerance of starch-loaded mice [12-15]. This also suggests that Cistanche deserticola has the effect of lowering blood sugar.

db / db Black mice are congenitally obese T2DM mice caused by defects in the leptin receptor gene. The leptin receptor gene loses function and hyperinsulinemia occurs within 2 weeks after birth. 3~ Obesity develops in 4 weeks, and very severe hyperglycemia develops in 8 weeks, accompanied by insulin resistance and β-cell failure [16-17]. Death usually occurs within 8 to 10 months and may be complicated by obvious renal failure. Disease［18］

. In this study, db/db black mice were used as experimental subjects to enrich the total polysaccharides, total oligosaccharides, and total glycosides of Cistanche deserticola and Tartschalis deserticola respectively. They conducted a comparative study on the hypoglycemic effects of different extracted parts. It is expected that Provide a scientific basis for the processing mechanism of Cistanche deserticola wine steaming.

1 Materials and instruments

1.1 Experimental animals BKS of spontaneous type 2 diabetes mouse model. Cg-Dock7m + / +Leprdb / JNju (genotype: db / db diabetes

Diseased black mouse) Male 7-week-old black mouse, body weight 35 ~ 40 g, DB / db m+ (genotype: db / m+ Black mouse) Male 7-week-old black mouse, body weight 18 ~ 20 g, from Provided by Nanjing University-Nanjing Institute of Biomedicine and Model Animal Institute of Nanjing University. This research plan was reviewed by the Experimental Animal Ethics Committee of Liaoning University of Traditional Chinese Medicine, and the approval number is 2018YSDW-030-02.

1.2 Drugs and reagents

The medicinal material Cistanche deserticola was collected from Alxa, Inner Mongolia (batch number: 20190508-1) in May 2019. It was identified as Cistanche desert (Cistanche desert) of the Orobanchiaceae plant by Professor Zhai Yanjun of the School of Pharmacy, Liaoning University of Traditional Chinese Medicine. Dry scaly leaves of cola Y. C. Ma) Fleshy stem, specimen preserved in Liaoning Provincial Traditional Chinese Medicine Processing Technology Industry Innovation Center.

Echinacoside reference substance (batch number: MUST -

13080801, purity ≥ 98%), verbascoside (verbascoside) reference substance (batch number: MUST-13122711, purity ≥ 98%), isomulascoside Glycoside (isoacteoside) reference substance (batch number: MUST-15081001, purity ≥99.77%), betaine reference substance (batch number: 894-200001, purity ≥98%) were purchased from China Institute for the Control of Pharmaceutical and Biological Products; D101 macroporous resin was provided by Chengdu Manster Biotechnology Co., Ltd.

Mouse anti-8-hydroxydeoxyguanosine (8-OHdG) monoclonal antibody [batch number: ab62623, Abcam (Shanghai) Trading Co., Ltd.]; PAS (batch number: G1281, Beijing Solebao Technology Co., Ltd.); anti-fluorescence Quenching mounting solution (Hai Biyuntian Biotechnology Co., Ltd.); SP9001 Rabbit SP detection kit (Wuxi Aorui Dongyuan Biotechnology Co., Ltd.); DAB, insulin (INS), glycosylated hemoglobin (HbA1c), triglyceride ( TG), mouse total cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), uric acid (UA), creatinine (Cr), malondialdehyde (MDA), superoxide Substance dismutase (SOD), urinary microalbumin

White (mALB) kit (Shanghai Langton Biotechnology Co., Ltd.). Cholesterol, xylene, absolute ethanol, acetonitrile, and methanol were of chromatographic grade, water was ultrapure water, and other reagents were of analytical grade.

1.3 Main instruments

  ACQUITY H-Class UPLC (Waters Company of the United States); blood glucose meter and blood glucose test strips (Bayer Healthcare Co., Ltd.); AE240 type 1/100,000 electronic analytical balance (Mettler, Switzerland) - Toledo Company); FA1004 electronic balance (Shanghai Precision Scientific Instrument Co., Ltd.); BN-518 automatic biological tissue embedding machine (Hubei Bernard Medical Technology Co., Ltd.); Electric constant temperature drying oven (Zhongwei Electronic Instrument Co., Ltd.); Leica RM2245 microtome (Leica Biosys, Germany) items); pH acidity meter (Shanghai Pengshun Scientific Instrument Co., Ltd.); Milli-Q Integral 3 type water purifier (French Molsheim Company).

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