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Libin Wang School of Medicine, Shaanxi Energy Institute, Xianyang, Shaanxi Province 712000, China

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Xin Shen Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing 100850, China

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Fang Wang School of Medicine, Shaanxi Energy Institute, Xianyang, Shaanxi Province 712000, China

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Xiaohui Xu Department of Orthopaedics, QILU Hospital of Shandong University Dezhou Hospital, DeZhou, Shandong Province 253000, China

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Abstract

Bai-Hu-Jia-Ren-Shen-Tang Decoction (BHJRSTD) is one of the oldest classic Chinese medicine prescriptions which used in the field of treatment of diabetes. However, to the best of our knowledge, the ingredients of this prescription have not been identified, and there are very few studies on the anti-diabetic mechanism of this prescription. Therefore, BHJRSTD was detected and identified by ultra-high-performance liquid chromatography coupled with Quadrupole-Exactive Focus Orbitrap MS (UHPLC–Q/Orbitrap/MS/MS). We identified 74 compounds, including flavonoids, alkaloids, chalcones, xanthones, phenols, phenylpropanoids, terpenes, triterpenes, amino acid derivatives, etc. Then, Sprague Dawley rats were fed with a high-fat and high-sugar diet for two months and injected with streptozotocin (STZ) to induce type 2 diabetes (T2DM). The diabetic rats were randomized to given metformin (200 mg kg−1·d−1, n = 15), BHJRSTD extracts (40 g kg−1·d−1) and BHJRSTD extracts (10 g kg−1·d−1) by gavage for 8 weeks. The results confirmed that BHJRSTD significantly decreased the level of MDA and increased levels of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), it shows that the prescription has significant antioxidant activity in the treatment of T2DM.

Abstract

Bai-Hu-Jia-Ren-Shen-Tang Decoction (BHJRSTD) is one of the oldest classic Chinese medicine prescriptions which used in the field of treatment of diabetes. However, to the best of our knowledge, the ingredients of this prescription have not been identified, and there are very few studies on the anti-diabetic mechanism of this prescription. Therefore, BHJRSTD was detected and identified by ultra-high-performance liquid chromatography coupled with Quadrupole-Exactive Focus Orbitrap MS (UHPLC–Q/Orbitrap/MS/MS). We identified 74 compounds, including flavonoids, alkaloids, chalcones, xanthones, phenols, phenylpropanoids, terpenes, triterpenes, amino acid derivatives, etc. Then, Sprague Dawley rats were fed with a high-fat and high-sugar diet for two months and injected with streptozotocin (STZ) to induce type 2 diabetes (T2DM). The diabetic rats were randomized to given metformin (200 mg kg−1·d−1, n = 15), BHJRSTD extracts (40 g kg−1·d−1) and BHJRSTD extracts (10 g kg−1·d−1) by gavage for 8 weeks. The results confirmed that BHJRSTD significantly decreased the level of MDA and increased levels of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), it shows that the prescription has significant antioxidant activity in the treatment of T2DM.

Introduction

In recent years, the advantages of Herbal and Traditional Chinese Medicine in the prevention and treatment of diabetes mellitus (DM) have been widely recognized around the world [12]. Bai-Hu-Jia-Ren-Shen-Tang Decoction (BHJRSTD), composed of Ginseng Radix, Rhizoma Anemarrhenae, Radix Glycyrrhizae Preparata, Gypsum Fibrosum and japonica rice is a traditional Chinese medicine prescriptions (TCMP) described in the Chinese medicine book “Treatise on Febrile and Miscellaneous Diseases”, which has been used in China for over 1,800 years [3]. The indications of BHJRSTD include infectious diseases, such as influenza, lung infection, encephalitis, enteric typhoid and hospital infection; thermoplegia; acute cerebrovascular disease, severe hyperosmolarity, diabetes mellitus [4], intractable hypotension, hypernatremia, shock and other internal diseases. This TCMP has been widely used to treat acute and severe cases in the cardiovascular intensive care ward. Refractory hypotension and hypovolemic shock that need large dose of supplemental fluid to maintain blood pressure and may also belong to the extension of the BHJRSTD formula syndrome.

Insulin resistance (IR) and pancreatic β-cell function defects were considered to be the main mechanisms of the pathogenesis of T2DM [5]. Oxidative stress can directly or indirectly aggravate IR and the damage of pancreatic β-cells, which is closely associated with the occurrence and development of T2DM [6, 7]. Studies have shown that compounds with antioxidant properties are more effective in treating T2DM [8, 9]. Nowadays, BHJRSTD is frequently used clinically in DM, especially as a complementary therapy [4]. The application of BHJRSTD proves to be more effective in the treatment of type 2 diabetes mellitus (T2DM) than using routine treatment only.

To our knowledge, there are no reports on chemical characterization and antioxidant activity studies of BHJRSTD, which is the usual way of consumption in traditional medicine in China. The complexity of chemicals in TCMP presents significant challenges to rapidly identify and characterize components. Over the past few decades, many natural product extracts, botanical ingredients and TCMP are analyzed using quadrupole Orbitrap spectrometry [10–12]. In this study, the main goals are develop an efficient method for rapid chemical characterization using UHPLC-Q-Exactive Orbitrap MS and measurement of the antioxidant activity of the BHJRSTD in diabetic rats, which are very beneficial for the further research and application of this TCMP including the material basis and pharmacological activity study.

Experimental

Materials

Streptozotocin (STZ) was MP Biomedicals, Trisodium citrate and citric acid were purchased from Sigma Aldrich Inc. (St Louis, MO, USA). Enzyme-linked immune adsorbent assay (ELISA) kits of Malondialdehyde (MDA), glutathione peroxidase (GSHPx), superoxide dismutase (SOD) and catalase (CAT) were obtained from R&D Systems Inc. (Minneapolis, MN, USA). Ginseng Radix, Rhizoma Anemarrhenae, Radix Glycyrrhizae Preparata, Gypsum Fibrosum and japonica rice were purchased from Xi'an Grass Plant Technology Co. Ltd. (Xi'an, Shaanxi, China). Methanol and acetonitrile of LC–MS grade were purchased from Fisher (Fisher Corp., Fair lawn, NJ, USA). All other chemicals were of analytical grade available commercially. ALL Chinese herbal medicine were obtained from Department of Chinese Materia Medica and Natural Medicines, School of Pharmacy, Fourth Military Medical University, and botanically identified by Professor Xiao-mei Song a traditional Chinese Medicine chemist at College of pharmacy, Shaanxi University of Chinese Medicine.

Preparation of BHJRSTD extracts

Ginseng Radix (10 g), Rhizoma Anemarrhena (18 g), Radix Glycyrrhizae Preparata (6 g), Gypsum Fibrosum (50 g) and japonica rice (9 g) were mixed in a classical dosage ratio used in the Han Dynasty. The mixture was soaked in distilled water (750 mL) for 30 min in a beaker (2,000 mL), then refluxed at 100 °C for 1 h twice under nitrogen protection in a round-bottomed flask (3,000 mL). The extraction solutions were combined and to be filtered, partial moisture was removed by rotary evaporator under reduced pressure. Then, the residuary solution was condensed to crude drug (2 g ml−1), the extracts were stored at 4 °C and were brought to room temperature before use [8].

Sample preparation

1 mL of BHJRSTD extracts was vortexed for 30 s at 4 °C and centrifuged at 12,000 rpm for 15 min. Then take 200 μL of supernatant into an EP tube, added 1 mL of extract (methanol: water = 4:1); Vortexed for 30 s and sonicated in ice-water bath for 5 min; After standing at −40 °C for 1 h, the samples were centrifuged at 4 °C and 12,000 rpm for 15 min; The supernatant was carefully filtered through a 0.22 μm microporous filter and stored at −20 °C until assayed.

Instruments and conditions

LC-MS/MS analysis was performed on an Agilent ultra-high performance liquid chromatography 1290 UPLC system with a Waters UPLC BEH C18 column (1.7 μm, 2.1 × 100 mm). The flow rate was set at 0.4 mL min−1 and the sample injection volume was set at 5 μL. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The multi-step linear elution gradient program was as follows: 0–3.5 min, 95–85% A; 3.5–6 min, 85–70% A; 6–6.5 min, 70–70% A; 6.5–12 min, 70–30% A; 12–12.5 min, 30–30% A; 12.5–18 min, 30–0% A; 18–25 min, 0–0% A; 25–26 min, 0–95% A; 26–30 min, 95–95% A.

An Q Exactive Focus mass spectrometer coupled with an Xcalibur software was employed to obtain the MS and MS/MS data based on the IDA acquisition mode. During each acquisition cycle, the mass range was from 100 to 1,500, and the top three of every cycle were screened and the corresponding MS/MS data were further acquired. Sheath gas flow rate: 45 Arb, Aux gas flow rate: 15 Arb, Capillary temperature: 400 °C, Full ms resolution: 70,000, MS/MS resolution: 17,500, Collision energy: 15/30/45 in NCE mode, Spray Voltage: 4.0 kV (positive) or −3.6 kV (negative).

Animals and treatment

All the studies on animals were conducted in strict compliance in accordance with the Guide for the Care and Use of Laboratory Animals. The experimental protocol (2021-9-11) involving animals was reviewed and approved by the Institutional Animal Care and Use Committee of the Fourth Military Medical University. One-month-old, healthy, specific pathogen-free, Sprague Dawley rats weighing 120 ± 10 g were supplied by the Experimental Animal Center of Air Force Military Medical University. All rats were housed in a tidy animal room with constant temperature (23 °C ± 2 °C) and humidity (55% ± 10%), and a 12-h light/dark cycle. The rats had unrestricted access to water and food. After a week of feeding, all rats were randomly divided into two groups: (1) control group (n = 15): fed with normal diet and water; (2) model group (n = 60): fed with high-fat and high-sugar feed for two month. After these, model group rats were starved overnight and injected 40 mg kg−1 streptozotocin (STZ) to induce diabetes, and the control group was injected with an equal volume of citric acid-sodium citrate buffer solution (pH = 4.2). Seventy-two hours after injection, fasting for 12 h, blood glucose was measured, after one week, a re-examination was performed 1 week later. The fasting blood glucose (FBG) was greater than 11.1 mmol L−1 twice, and then the T2DM model was considered successfully established. T2DM rats were randomly divided into 4 groups: (1) T2DM (DM) model group (n = 15) (2) metformin (Met) group (200 mg kg−1·d−1, n = 15); (3) high-dose (H) BHJRSTD group (40 g kg−1·d−1, n = 15); (4) low-dose (L) BHJRSTD group (10 g kg−1·d−1, n = 15). The model groups were given high-fat and high-sugar diet every day, and the normal control (NC) group continued to be given the normal diet and water. Each group was administration through gastric gavage for one month to verify the hypoglycemic effect. Body weight, food consumption, and FBG levels were monitored and recorded during the administration period.

Collection and processing of test samples

After 8 weeks of administration, then all rats were killed, the excised pancreas tissues from the normal and experimental rats were immediately rinsed with normal saline and fixed in 4% paraformaldehyde. The tissues were cut into 5 μm thick sections after embedding in paraffin and then stained with hematoxylin/eosin staining (HE) for histopathological examination.

Antioxidant activity

The CAT, SOD, GSH-Px are important ones of antioxidant defense systems. MDA is formed as an end product of lipid peroxidation. Briefly, portions of pancreatic tissue were homogenized in cool phosphate-buffered saline (pancreatic tissue to normal saline, 1:4). The levels of CAT, SOD, GSH-Px and MDA in pancreatic tissue was measured using a commercially available ELISA kit, according to the manufacturer's instructions.

Statistical analysis

The results of the antioxidant assays were statistically processed using GraphPad Prism 6.0 (P < 0.05; Dunnett's test) to determine significant differences between multiple groups.

Results and discussion

Identification of BHJRSTD extracts

To comprehensively characterize the BHJRSTD extract, an analytical method based on UHPLC Q-Exactive Focus Orbitrap MS was established in this research. Firstly, the prepared sample was injected into the UHPLC Q-Exactive Focus Orbitrap MS to obtain full-scan high-resolution MS data. Then, these data were processed using the Q Exactive Focus mass spectrometer's control software (Xcalibur, Thermo Fisher Scientific) to acquire, detect and predict various types of molecules. Third, the MS2 of the predicted molecule was obtained in parallel reaction monitoring mode via UHPLC Q-Exactive Focus Orbitrap MS. Finally, compounds were identified by full scan MS date, MS2 data, retention times and references. Details of the 74 major compounds are shown in Table 1 and Fig. 1.

Table 1.

High resolution UHPLC–Q/Orbitrap/MS/MS identification of ingredients from BHJRSTD extracts

Peak no. Components name Formula InChIKey RT (min) Measured Mass (m/z) Quasi-Molecular Ion (m/z) Class
1 Isoleucine C6H13NO2 AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.75 132.1020527 132.102

[M+H]+
Amino acid
2 L-Proline C5H9NO2 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.75 116.0706856 116.071

[M+H]+
Amino acid
3 Nicotinic acid C6H5NO2 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.87 124.0394663 124.039

[M+H]+
Vitamin
4 L-Isoleucine C6H13NO2 AGPKZVBTJJNPAG-WHFBIAKZSA-N 1.03 132.1020398 132.102

[M+H]+
Amino acid
5 L-Tryptophan C11H12N2O2 QIVBCDIJIAJPQS-VIFPVBQESA-N 2.34 205.0974881 205.097

[M+H]+
Organoheterocyclic compounds
6 Neomangiferin C25H28O16 VUWOVGXVRYBSGI-IRXABLMPSA-N 2.62 585.1453811 585.145

[M+H]+
Xanthones
7 Mangiferin C19H18O11 AEDDIBAIWPIIBD-ZJKJAXBQSA-N 3.07 423.092836 423.093

[M+H]+
Xanthones
8 Isomangiferin C19H18O11 CDYBOKJASDEORM-HBVDJMOISA-N 4.76 423.0929111 423.093

[M+H]+
Xanthones
9 Liquiritin C21H22O9 DEMKZLAVQYISIA-UZQFATADSA-N 5.30 441.114928 441.115

[M+Na]+
Flavonoids
10 Liquiritigenin C15H12O4 FURUXTVZLHCCNA-AWEZNQCLSA-N 5.96 257.0808213 257.081

[M+H]+
Flavonoids
11 Licochalcone B C16H14O5 DRDRYGIIYOPBBZ-XBXARRHUSA-N 6.21 287.0911089 287.091

[M+H]+
Chalcones
12 Pseudoginsenoside F11 C42H72O14 JBGYSAVRIDZNKA-UHFFFAOYSA-N 7.01 801.5010963 801.501

[M+H]+
Terpenoids
13 Isoliquiritigenin C15H12O4 DXDRHHKMWQZJHT-FPYGCLRLSA-N 7.64 257.0808108 257.081

[M+H]+
Chalcones
14 Ginsenoside Rg1 C42H72O14 YURJSTAIMNSZAE-ILDRPQKSSA-N 8.40 823.4817834 823.482

[M+Na]+
Terpenoids
15 Kaempferide C16H12O6 SQFSKOYWJBQGKQ-UHFFFAOYSA-N 8.48 301.0710347 301.071

[M+H]+
Flavonoids
16 Ginsenoside Rg3 C42H72O13 RWXIFXNRCLMQCD-XXYKJBRESA-N 8.75 807.4859478 807.486

[M+Na]+
Terpenoids
17 Ginsenoside Rg5 C42H70O12 NJUXRKMKOFXMRX-UHFFFAOYSA-N 8.85 767.4953034 767.495

[M+H]+
Terpenoids
18 Glabrone C20H16O5 COLMVFWKLOZOOP-UHFFFAOYSA-N 9.11 337.1070099 337.107

[M+H]+
Flavonoids
19 Ginsenoside Re C48H82O18 PWAOOJDMFUQOKB-DQASNDRCSA-N 9.22 969.538058 969.538

[M+Na]+
Terpenoids
20 Licoricesaponin G2 C42H62O17 WBQVRPYEEYUEBQ-OJVDLISWSA-N 9.29 839.4050082 839.405

[M+H]+
Terpenoids
21 Ginsenoside Rf C42H72O14 UZIOUZHBUYLDHW-KIOKIGKZSA-N 9.59 801.4973674 801.497

[M+H]+
Terpenoids
22 Licoflavone A C20H18O4 HJGURBGBPIKRER-UHFFFAOYSA-N 10.35 323.1278005 323.128

[M+H]+
Flavonoids
23 Chikusetsu saponin IVa C42H66O14 YOSRLTNUOCHBEA-SNRHWUJASA-N 10.57 817.4358779 817.436

[M+Na]+
Terpenoids
24 Licochalcone A C21H22O4 KAZSKMJFUPEHHW-DHZHZOJOSA-N 10.76 339.1589659 339.159

[M+H]+
Flavonoids
25 Sarsasapogenin C27H44O3 GMBQZIIUCVWOCD-WWASVFFGSA-N 11.49 417.33681 417.337

[M+H]+
Terpenoids
26 Timosaponin A-III C39H64O13 MMTWXUQMLQGAPC-HRSJJQGESA-N 11.49 741.4428077 741.443

[M+H]+
Terpenoids
27 Ginsenoside Rk1 C42H70O12 KWDWBAISZWOAHD-MHOSXIPRSA-N 12.35 789.4744795 789.474

[M+Na]+
Terpenoids
28 20(S)-Ginsenoside Ck C36H62O8 FVIZARNDLVOMSU-IRFFNABBSA-N 12.60 645.4331453 645.433

[M+Na]+
Terpenoids
29 L-Valine C5H11NO2 KZSNJWFQEVHDMF-BYPYZUCNSA-N 26.14 118.0862706 118.086

[M+H]+
Amino acid
30 Cinnamic acid C9H8O2 WBYWAXJHAXSJNI-VOTSOKGWSA-N 2.58 147.0451477 147.045

[M−H]
Phenylpropanoids
31 Phenylacetic acid C8H8O2 WLJVXDMOQOGPHL-UHFFFAOYSA-N 2.63 135.045226 135.045

[M−H]
Aromaticity
32 Gallic acid C7H6O5 LNTHITQWFMADLM-UHFFFAOYSA-N 2.77 169.0144344 169.014

[M−H]
Phenols
33 Cryptochlorogenic acid C16H18O9 GYFFKZTYYAFCTR-AVXJPILUSA-N 3.16 353.0880346 353.088

[M−H]
Phenylpropanoids
34 2-Hydroxycinnamic acid, predominantly trans C9H8O3 PMOWTIHVNWZYFI-AATRIKPKSA-N 3.98 163.0402139 163.040

[M−H]
Phenylpropanoids
35 Daidzin C21H20O9 KYQZWONCHDNPDP-QNDFHXLGSA-N 4.45 461.1104935 461.110

[M−H]
Flavonoids
36 Homoorientin C21H20O11 ODBRNZZJSYPIDI-VJXVFPJBSA-N 4.58 447.0945323 447.095

[M−H]
Flavonoids
37 Gentiopicroside C16H20O9 DUAGQYUORDTXOR-UHFFFAOYNA-N 4.65 355.1038508 355.104

[M−H]
Iridoids
38 Gossypetin-8-C-glucoside C21H20O13 SJRXVLUZMMDCNG-UHFFFAOYSA-N 4.75 479.0832296 479.083

[M−H]
Flavonoids
39 Orientin C21H20O11 PLAPMLGJVGLZOV-VPRICQMDSA-N 4.76 447.0944571 447.094

[M−H]
Flavonoids
40 Pseudolaric acid B C23H28O8 VDGOFNMYZYBUDT-VAFVZNMKSA-N 4.77 431.1691084 431.169

[M−H]
Terpenoids
41 Eleutheroside E C34H46O18 FFDULTAFAQRACT-UHFFFAOYSA-N 5.00 787.2695132 787.270

[M+HCOO]
Phenylpropanoids
42 Vitexin C21H20O10 SGEWCQFRYRRZDC-VPRICQMDSA-N 5.22 431.098706 431.099

[M−H]
Flavonoids
43 Quercetin-3-O-galactoside C21H20O12 OVSQVDMCBVZWGM-UHFFFAOYSA-N 5.62 463.0886957 463.089

[M−H]
Flavonoids
44 Tectoridin C22H22O11 CNOURESJATUGPN-UDEBZQQRSA-N 5.87 461.1103949 461.110

[M−H]
Flavonoids
45 Digitonin C56H92O29 UVYVLBIGDKGWPX-UHFFFAOYNA-N 6.21 1227.566017 1227.566

[M−H]
Terpenoids
46 Glycitin C22H22O10 OZBAVEKZGSOMOJ-MIUGBVLSSA-N 6.44 445.1151963 445.115

[M−H]
Flavonoids
47 Isovitexin C21H20O10 MYXNWGACZJSMBT-VJXVFPJBSA-N 6.45 431.0985813 431.099

[M−H]
Flavonoids
48 Acacetin-7-O-rutinoside C28H32O14 YFVGIJBUXMQFOF-UHFFFAOYNA-N 6.47 591.1737251 591.174

[M−H]
Flavonoids
49 Isoliquiritin C21H22O9 YNWXJFQOCHMPCK-LXGDFETPSA-N 6.55 417.1195589 417.120

[M−H]
Chalcones
50 Timosaponin B II C45H76O19 SORUXVRKWOHYEO-UHFFFAOYSA-N 6.77 919.4922594 919.492

[M−H]
Terpenoids
51 Atractyloside A C21H36O10 QNBLVYVBWDIWDM-BSLJOXIBSA-N 7.03 493.2290177 493.229

[M+HCOO]
Terpenoids
52 Hesperetin C16H14O6 AIONOLUJZLIMTK-AWEZNQCLSA-N 7.21 301.0723311 301.072

[M−H]
Flavonoids
53 Ginsenoside F3 C41H70O13 HJRVLGWTJSLQIG-UHFFFAOYSA-N 7.49 815.4799289 815.480

[M+HCOO]
Terpenoids
54 Terrestrosin K C51H82O24 TVRRDUXJKROMDX-CXLJPAHDSA-N 7.68 1077.512784 1077.513

[M−H]
Triterpenoids
55 Naringenin C15H12O5 FTVWIRXFELQLPI-UHFFFAOYNA-N 7.92 271.0614345 271.061

[M−H]
Flavonoids
56 Ginsenoside Rh1 C36H62O9 RAQNTCRNSXYLAH-UHFFFAOYSA-N 7.99 683.4375706 683.438

[M+HCOO]
Terpenoids
57 Kaempferol C15H10O6 IYRMWMYZSQPJKC-UHFFFAOYSA-N 8.10 285.0404805 285.040

[M−H]
Flavonoids
58 Ginsenoside Rb1 C54H92O23 GZYPWOGIYAIIPV-UHFFFAOYNA-N 8.54 1153.599607 1153.600

[M+HCOO]
Terpenoids
59 Ginsenoside Rc C53H90O22 JDCPEKQWFDWQLI-UHFFFAOYSA-N 8.67 1077.587843 1077.588

[M−H]
Terpenoids
60 Ginsenoside Rg2 C42H72O13 AGBCLJAHARWNLA-UHFFFAOYNA-N 8.75 783.4898787 783.490

[M−H]
Terpenoids
61 Ginsenoside Ro C48H76O19 NFZYDZXHKFHPGA-QQHDHSITSA-N 8.80 955.4926865 955.493

[M−H]
Terpenoids
62 Ginsenoside Rb2 C53H90O22 NODILNFGTFIURN-UHFFFAOYNA-N 8.85 1123.595118 1123.595

[M+HCOO]
Terpenoids
63 Ginsenoside F1 C36H62O9 XNGXWSFSJIQMNC-UHFFFAOYNA-N 8.96 683.4385504 683.439

[M+HCOO]
Terpenoids
64 Araloside A C47H74O18 KQSFNXMDCOFFGW-UGBTYEQWSA-N 9.03 925.4819032 925.482

[M−H]
Terpenoids
65 3-[(Carboxycarbonyl)amino]-l-alanine C5H8N2O5 NEEQFPMRODQIKX-REOHCLBHSA-N 9.43 174.9563911 174.956

[M−H]
Alkaloids
66 Isoanhydroicaritin C21H20O6 RJANATGWWPNKAL-UHFFFAOYSA-N 9.79 367.1197283 367.120

[M−H]
Flavonoids
67 Icaritin C21H20O6 TUUXBSASAQJECY-UHFFFAOYSA-N 10.24 367.1197729 367.120

[M−H]
Flavonoids
68 Neobavaisoflavone C20H18O4 OBGPEBYHGIUFBN-UHFFFAOYSA-N 10.36 321.114183 321.114

[M−H]
Flavonoids
69 xanthohumol C21H22O5 ORXQGKIUCDPEAJ-YRNVUSSQSA-N 10.57 353.1398336 353.140

[M−H]
Flavonoids
70 Inermin C16H12O5 HUKSJTUUSUGIDC-ZBEGNZNMSA-N 10.64 283.0616985 283.062

[M−H]
Flavonoids
71 Ginsenoside Rg3(S-FORM) C42H72O13 RWXIFXNRCLMQCD-UHFFFAOYNA-N 10.96 783.4903466 783.490

[M−H]
Terpenoids
72 Momordin Ic C41H64O13 HWYBGIDROCYPOE-WEAQAMGWSA-N 11.14 763.4297182 763.430

[M−H]
Terpenoids
73 Glabridin C20H20O4 LBQIJVLKGVZRIW-ZDUSSCGKSA-N 11.32 323.1290457 323.129

[M−H]
Flavonoids
74 Mulberrin C25H26O6 UWQYBLOHTQWSQD-UHFFFAOYSA-N 13.01 421.1659774 421.166

[M−H]
Flavonoids
Fig. 1.
Fig. 1.

UHPLC–Q/Orbitrap/MS/MS chromatograms of BHJRSTD extracts (total ion current; a: positive ion mode; b: negative ion mode)

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01068

Peaks 9, 10, 15, 18, 22, 24, 35, 36, 38, 39, 42–44, 46–48, 52, 55, 57, 66–70, 73 and 74 were identified as flavonoids based on comparing the retention time, measured mass, and MS spectrum with reference standards. Peaks 5 and 65 (Alkaloids) were identified as L-Tryptophan and 3-[(Carboxycarbonyl)amino]-L-alanine, respectively. Peaks 2 and 4 with the same [M+H]+ ion at m/z 116.071, 132.102 were identified as amino acid. Peak 31 was identified as phenylacetic acid and peak 37 was identified as gentiopicroside. Peak 11, 13, 49 were identified as chalcone. Peak 32 was identified as gallic acid (Phenols). Peak 30, 33, 34, 41 were regarded as phenylpropanoids. Peak 12, 14, 16, 17, 19–21, 23, 25–28, 40, 45, 50, 51, 53, 56, 58–64, 71, 72 were identified as terpenoids, while peak 54 was identified as terrestrosin K (Triterpenoids). Peak 6–8 (Xanthones) were identified as neomangiferin, mangiferin and isomangiferin, respectively.

Effects of BHJRSTD on the histomorphology of pancreatic islets in T2DM rats

As shown in Fig. 2, the pancreatic islets in the NC group were oval in shape, with regular structure, different size of cell clusters, regular shape, complete structure, clear edge, and scattered among the pancreatic vesicles. The β-cells in pancreatic islets were abundant, plentiful in shape, uniform in size, and tightly arranged, no pathological changes were found. In contrast, the shape of pancreatic islets in the DM group was blurred, irregular, and structurally disordered. The β-cells in the pancreatic islets were deformed and swollen, and the cytoplasm was lightly stained. Compared with the pancreatic islets of the DM group, the number of islet β-cells in the MET group, H group and L group was significantly increased, meanwhile, the structure of β-cells was regular and no obvious necrosis was seen. The islet borders were clear and the morphological structure was improved.

Fig. 2.
Fig. 2.

Hematoxylin and eosin staining of T2DM rats pancreatic tissue

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01068

Effects of BHJRSTD on the level of oxidative stress in T2DM rats

The human body is constantly exposed to different types of agents, resulting in the production of reactive species called free radicals [13], which lead to the oxidation of cellular machinery through the transfer of their free unpaired electrons [14]. In response to the harmful effects of these species, the body has acquired an endogenous antioxidant system or obtained exogenous antioxidants from the diet to neutralize these species and maintain the body's homeostasis. Any imbalance between reactive species and antioxidants might cause a condition called “oxidative stress”, which can lead to the development of pathological conditions, one of which is diabetes [15]. The damage of these free radicals cause to cells can be quantified by measuring levels of MDA, a product of lipid peroxidation [16, 17]. Certain enzymes play important roles in antioxidant defense to maintain viable reproductive capacity. These enzymes include CAT, SOD and GSH-Px, which convert free radicals or reactive oxygen intermediates to non-radical products [18].

In this study, the effect of BHJRSTD on the level of anti-oxidative stress in T2DM rats was evaluated via observing the contents of CAT, SOD, MDA and GSH-Px in rat pancreatic tissue. Figure 3 illustrates that, compared with the NC group, the levels of CAT, SOD, and GSH-Px in the DM group were significantly decreased (P < 0.05), and the level of MDA was significantly increased (P < 0.05). Compared with the DM group, the levels of CAT, SOD and GSH-Px were significantly decreased (P < 0.05), and the level of MDA in the MET group was significantly increased (P < 0.05). After administration of BHJRSTD extracts to rats for 8 weeks, the levels of SOD and GSH-Px in H and L groups were significantly increased (P < 0.05). Compared with the DM group, H and L groups showed an increasing trend, but the difference was not statistically significant (P > 0.05). BHJRSTD could reduce the MDA level in H and L groups, and there was a significant difference compared with the DM group (P < 0.05). The results indicated that BHJRSTD extracts had a certain anti-oxidative stress effect on pancreatic tissue of T2DM rats.

Fig. 3.
Fig. 3.

The effects of each group on the level of oxidative stress in T2DM rats (#P < 0.05 vs NC group, *P < 0.05 vs DM group; Mean ± SD, n = 8)

Citation: Acta Chromatographica 2022; 10.1556/1326.2022.01068

Conclusion

In the present investigation, the potential chemical composition and antioxidant effect of BHJRSTD extracts were evaluated via UHPLC–Q/Orbitrap/MS/MS and ELISA kit. We identified 74 compounds, including flavonoids, alkaloids, chalcones, xanthones, phenols, phenylpropanoids, terpenes, triterpenes, terpenes, amino acid derivatives, etc. It obviously and increased the content of CAT, SOD, GSH-Px and reduced level of MDA in T2DM rats after 8 weeks of administration. This study provides some reference information for the clinical use of the BHJRSTD, and also provides some basis for the development of Chinese herbal medicine

Acknowledgement

This research was supported by the Scientific Research Program Funded by Education Department of Shaanxi Provincial Government, Shaanxi Province, China (Grant No: 21JK0578) and Basic Research plan of Natural Science of Shaanxi Province, China (Grant No: 2021JQ-887, 2021JQ-888).

References

  • 1.

    Xu, Y. X. Z. ; Xi, S. ; Qian, X. Evaluating traditional Chinese medicine and herbal products for the treatment of gestational diabetes mellitus. J. Diabetes Res. 2019, 2019, 9182595.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Wang, J. ; Ma, Q. ; Li, Y. ; Wang, M. ; Wang, T. ; Zhao, B. Research progress on Traditional Chinese Medicine syndromes of diabetes mellitus. Biomed. Pharmacother. 2020, 121, 109565.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 3.

    Liu, H.-K. ; Hung, T.-M. ; Huang, H.-C. ; Huang, H. C. ; Lee, I. ; Chang, C. C. ; Cheng, J. J. ; Huang, C. Bai-Hu-Jia-Ren-Shen-Tang decoction reduces fatty liver by activating AMP-activated protein kinase in vitro and in vivo. Evidence-Based Complement. Altern. Med. 2015, 2015, 651734.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4.

    Tian, Y. ; Zhong, W. ; Zhang, Y. ; Zhou, L. ; Fu, X. ; Wang, L. ; Xie, C. Baihu Jia Renshen Decoction for type 2 diabetic mellitus: a protocol for systematic review and meta-analysis. Medicine (Baltimore) 2020, 99(19) e20210–e20210.

    • Search Google Scholar
    • Export Citation
  • 5.

    Abdul-Ghani, M. A. ; Tripathy, D. ; DeFronzo, R. A. Contributions of β-cell dysfunction and insulin resistance to the pathogenesis of impaired glucose tolerance and impaired fasting glucose. Diabetes Care 2006, 29(5), 11301139.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Caimi, G. ; Carollo, C. ; Presti, R. L. Diabetes mellitus: oxidative stress and wine. Curr. Med. Res. Opin. 2003, 19(7), 581586.

  • 7.

    Patil, V. S. ; Patil, V. P. ; Gokhale, N. ; Acharya, A. ; Kangokar, P. Chronic periodontitis in type 2 diabetes mellitus: oxidative stress as a common factor in periodontal tissue injury. J. Clin. Diagn. Res. 2016, 10(4), BC12BC16.

    • Search Google Scholar
    • Export Citation
  • 8.

    Unuofin, J. O. ; Lebelo, S. L. Antioxidant effects and mechanisms of medicinal plants and their bioactive compounds for the prevention and treatment of type 2 diabetes: an updated review. Oxidative Med. Cell Longevity 2020, 2020, 1356893.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Dembinska-Kiec, A. ; Mykkänen, O. ; Kiec-Wilk, B. ; Mykkänen, H. Antioxidant phytochemicals against type 2 diabetes. Br. J. Nutr. 2008, 99(E-S1), ES109ES117.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Qiao, X. ; Li, R. ; Song, W. ; Miao, W. J. ; Liu, J. ; Chen, H. B. ; Ye, M. A targeted strategy to analyze untargeted mass spectral data: rapid chemical profiling of Scutellaria baicalensis using ultra-high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry and key ion filtering. J. Chromatogr. A 2016, 1441, 8395.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Nardin, T. ; Piasentier, E. ; Barnaba, C. ; Larcher, R. Targeted and untargeted profiling of alkaloids in herbal extracts using online solid-phase extraction and high-resolution mass spectrometry (Q-Orbitrap). J. Mass Spectrom. 2016, 51(9), 729741.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Gómez, J. ; Simirgiotis, M. J. ; Lima, B. ; Paredes, J. D. ; Villegas Gabutti, C. M. ; Gamarra-Luques, C. ; Tapia, A. Antioxidant, gastroprotective, cytotoxic activities and UHPLC PDA-Q orbitrap mass spectrometry identification of metabolites in baccharis grisebachii decoction. Molecules 2019, 24(6), 1085.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Feinendegen, L. E. Reactive oxygen species in cell responses to toxic agents. Hum. Exp. Toxicol. 2002, 21(2), 8590.

  • 14.

    Asmat, U. ; Abad, K. ; Ismail, K. Diabetes mellitus and oxidative stress—a concise review. Saudi Pharm. J. 2016, 24(5), 547553.

  • 15.

    Wei, W. ; Liu, Q. ; Tan, Y. ; Liu, L. ; Li, X. ; Cai, L. Oxidative stress, diabetes, and diabetic complications. Hemoglobin 2009, 33(5), 370377.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Tiwari, B. K. ; Pandey, K. B. ; Abidi, A. B. ; Rizvi, S. I. Markers of oxidative stress during diabetes mellitus. J. Biomarkers 2013, 2013, 378790.

  • 17.

    Yaribeygi, H. ; Sathyapalan, T. ; Atkin, S. L. ; Sahebkar, A. Molecular mechanisms linking oxidative stress and diabetes mellitus. Oxidative Med. Cell Longevity 2020, 2020, 8609213.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Djordjevic, A. ; Spasic, S. ; Jovanovic-Galovic, A. ; Djordjevic, R. ; Grubor-Lajsic, G. Oxidative stress in diabetic pregnancy: SOD, CAT and GSH-Px activity and lipid peroxidation products. J. Maternal-Fetal Neonatal Med. 2004, 16(6), 367372.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 1.

    Xu, Y. X. Z. ; Xi, S. ; Qian, X. Evaluating traditional Chinese medicine and herbal products for the treatment of gestational diabetes mellitus. J. Diabetes Res. 2019, 2019, 9182595.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Wang, J. ; Ma, Q. ; Li, Y. ; Wang, M. ; Wang, T. ; Zhao, B. Research progress on Traditional Chinese Medicine syndromes of diabetes mellitus. Biomed. Pharmacother. 2020, 121, 109565.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 3.

    Liu, H.-K. ; Hung, T.-M. ; Huang, H.-C. ; Huang, H. C. ; Lee, I. ; Chang, C. C. ; Cheng, J. J. ; Huang, C. Bai-Hu-Jia-Ren-Shen-Tang decoction reduces fatty liver by activating AMP-activated protein kinase in vitro and in vivo. Evidence-Based Complement. Altern. Med. 2015, 2015, 651734.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4.

    Tian, Y. ; Zhong, W. ; Zhang, Y. ; Zhou, L. ; Fu, X. ; Wang, L. ; Xie, C. Baihu Jia Renshen Decoction for type 2 diabetic mellitus: a protocol for systematic review and meta-analysis. Medicine (Baltimore) 2020, 99(19) e20210–e20210.

    • Search Google Scholar
    • Export Citation
  • 5.

    Abdul-Ghani, M. A. ; Tripathy, D. ; DeFronzo, R. A. Contributions of β-cell dysfunction and insulin resistance to the pathogenesis of impaired glucose tolerance and impaired fasting glucose. Diabetes Care 2006, 29(5), 11301139.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Caimi, G. ; Carollo, C. ; Presti, R. L. Diabetes mellitus: oxidative stress and wine. Curr. Med. Res. Opin. 2003, 19(7), 581586.

  • 7.

    Patil, V. S. ; Patil, V. P. ; Gokhale, N. ; Acharya, A. ; Kangokar, P. Chronic periodontitis in type 2 diabetes mellitus: oxidative stress as a common factor in periodontal tissue injury. J. Clin. Diagn. Res. 2016, 10(4), BC12BC16.

    • Search Google Scholar
    • Export Citation
  • 8.

    Unuofin, J. O. ; Lebelo, S. L. Antioxidant effects and mechanisms of medicinal plants and their bioactive compounds for the prevention and treatment of type 2 diabetes: an updated review. Oxidative Med. Cell Longevity 2020, 2020, 1356893.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Dembinska-Kiec, A. ; Mykkänen, O. ; Kiec-Wilk, B. ; Mykkänen, H. Antioxidant phytochemicals against type 2 diabetes. Br. J. Nutr. 2008, 99(E-S1), ES109ES117.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Qiao, X. ; Li, R. ; Song, W. ; Miao, W. J. ; Liu, J. ; Chen, H. B. ; Ye, M. A targeted strategy to analyze untargeted mass spectral data: rapid chemical profiling of Scutellaria baicalensis using ultra-high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry and key ion filtering. J. Chromatogr. A 2016, 1441, 8395.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Nardin, T. ; Piasentier, E. ; Barnaba, C. ; Larcher, R. Targeted and untargeted profiling of alkaloids in herbal extracts using online solid-phase extraction and high-resolution mass spectrometry (Q-Orbitrap). J. Mass Spectrom. 2016, 51(9), 729741.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Gómez, J. ; Simirgiotis, M. J. ; Lima, B. ; Paredes, J. D. ; Villegas Gabutti, C. M. ; Gamarra-Luques, C. ; Tapia, A. Antioxidant, gastroprotective, cytotoxic activities and UHPLC PDA-Q orbitrap mass spectrometry identification of metabolites in baccharis grisebachii decoction. Molecules 2019, 24(6), 1085.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Feinendegen, L. E. Reactive oxygen species in cell responses to toxic agents. Hum. Exp. Toxicol. 2002, 21(2), 8590.

  • 14.

    Asmat, U. ; Abad, K. ; Ismail, K. Diabetes mellitus and oxidative stress—a concise review. Saudi Pharm. J. 2016, 24(5), 547553.

  • 15.

    Wei, W. ; Liu, Q. ; Tan, Y. ; Liu, L. ; Li, X. ; Cai, L. Oxidative stress, diabetes, and diabetic complications. Hemoglobin 2009, 33(5), 370377.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Tiwari, B. K. ; Pandey, K. B. ; Abidi, A. B. ; Rizvi, S. I. Markers of oxidative stress during diabetes mellitus. J. Biomarkers 2013, 2013, 378790.

  • 17.

    Yaribeygi, H. ; Sathyapalan, T. ; Atkin, S. L. ; Sahebkar, A. Molecular mechanisms linking oxidative stress and diabetes mellitus. Oxidative Med. Cell Longevity 2020, 2020, 8609213.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Djordjevic, A. ; Spasic, S. ; Jovanovic-Galovic, A. ; Djordjevic, R. ; Grubor-Lajsic, G. Oxidative stress in diabetic pregnancy: SOD, CAT and GSH-Px activity and lipid peroxidation products. J. Maternal-Fetal Neonatal Med. 2004, 16(6), 367372.

    • Crossref
    • Search Google Scholar
    • Export Citation
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Senior editors

Editor(s)-in-Chief: Kowalska, Teresa

Editor(s)-in-Chief: Sajewicz, Mieczyslaw

Editors(s)

  • Danica Agbaba (University of Belgrade, Belgrade, Serbia)
  • Ivana Stanimirova-Daszykowska (University of Silesia, Katowice, Poland)
  • Monika Waksmundzka-Hajnos (Medical University of Lublin, Lublin, Poland)

Editorial Board

  • R. Bhushan (The Indian Institute of Technology, Roorkee, India)
  • J. Bojarski (Jagiellonian University, Kraków, Poland)
  • B. Chankvetadze (State University of Tbilisi, Tbilisi, Georgia)
  • M. Daszykowski (University of Silesia, Katowice, Poland)
  • T.H. Dzido (Medical University of Lublin, Lublin, Poland)
  • A. Felinger (University of Pécs, Pécs, Hungary)
  • K. Glowniak (Medical University of Lublin, Lublin, Poland)
  • B. Glód (Siedlce University of Natural Sciences and Humanities, Siedlce, Poland)
  • A. Gumieniczek (Medical University of Lublin, Lublin, Poland)
  • U. Hubicka (Jagiellonian University, Kraków, Poland)
  • K. Kaczmarski (Rzeszow University of Technology, Rzeszów, Poland)
  • H. Kalász (Semmelweis University, Budapest, Hungary)
  • K. Karljiković Rajić (University of Belgrade, Belgrade, Serbia)
  • I. Klebovich (Semmelweis University, Budapest, Hungary)
  • A. Koch (Private Pharmacy, Hamburg, Germany)
  • Ł. Komsta (Medical University of Lublin, Lublin, Poland)
  • P. Kus (Univerity of Silesia, Katowice, Poland)
  • D. Mangelings (Free University of Brussels, Brussels, Belgium)
  • E. Mincsovics (Corvinus University of Budapest, Budapest, Hungary)
  • Á. M. Móricz (Centre for Agricultural Research, Budapest, Hungary)
  • G. Morlock (Giessen University, Giessen, Germany)
  • A. Petruczynik (Medical University of Lublin, Lublin, Poland)
  • R. Skibiński (Medical University of Lublin, Lublin, Poland)
  • B. Spangenberg (Offenburg University of Applied Sciences, Germany)
  • T. Tuzimski (Medical University of Lublin, Lublin, Poland)
  • Y. Vander Heyden (Free University of Brussels, Brussels, Belgium)
  • A. Voelkel (Poznań University of Technology, Poznań, Poland)
  • B. Walczak (University of Silesia, Katowice, Poland)
  • W. Wasiak (Adam Mickiewicz University, Poznań, Poland)
  • I.G. Zenkevich (St. Petersburg State University, St. Petersburg, Russian Federation)

 

KOWALSKA, TERESA
E-mail: kowalska@us.edu.pl

SAJEWICZ, MIECZYSLAW
E-mail:msajewic@us.edu.pl

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