Restoration of gut dysbiosis through Clostridium butyricum and magnesium possibly balance blood glucose levels: an experimental study

doi:10.21203/rs.3.rs-3339607/v1

Download PDF

Research Article

Restoration of gut dysbiosis through Clostridium butyricum and magnesium possibly balance blood glucose levels: an experimental study

https://doi.org/10.21203/rs.3.rs-3339607/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 01 Apr, 2024

Read the published version in BMC Microbiology →

You are reading this latest preprint version

Diabetes mellitus (DM) is a chronic metabolic disorder characterized by an elevated level of blood glucose due to the absence of insulin secretion, ineffectiveness, or lack of uptake of secreted insulin in the body. The improperly diagnosed and poorly managed DM can cause severe damage to organs in the body like the nerves, eyes, heart, and kidneys. This study was aimed at investigating the effect of probiotic Clostridium butyricum with magnesium supplementation to evaluate the effect on gut microbial dysbiosis and blood glucose levels. In an animal house, 24 male albino rats weighing 200–250 g have free access to water and food. Diabetes was induced using streptozotocin 60 mg/kg in overnight fasted rats. Diabetic rats were divided into four groups (n = 6). Control group (G₀) rats were given Metformin 100 mg/kg/day with a standard basal diet, treatment group 1 was given Clostridium Butyricum 2.5x10⁵ CFU/day with standard basal diet, magnesium 500 mg/kg/day to group 2, While Clostridium Butyricum (1.5x10⁵ CFU/day) and magnesium (300 mg/kg/day) in combination with a standard basal diet to group 3. Random blood glucose levels were monitored twice a week for 21 days and were represented as mean of each week. The results conclude that Clostridium Butyricum 2.5 x10⁵ CFU is very effective in balancing random blood glucose levels from 206.6 ± 67.7 to 85.1 ± 3.8 (p = 0.006) compared to other groups (p > 0.005). The results of stool analysis showed that Clostridium Butyricum restores microbial dysbiosis as evident by the 10⁵ CFU Clostridium Butyricum load in G₁, which was higher than G₀, G₂ and G₃ which were 10³ and 10⁴ CFU respectively. The findings of this study conclude that Clostridium Butyricum supplementation improved blood glucose levels and intestinal bacterial load in type II diabetes mellitus.

Diabetes Mellitus

Probiotic

Magnesium Supplementation

Clostridium Butyricum

Diabetes mellitus (DM) is a chronic metabolic disorder characterized by hyperglycemia due to either the absence of insulin secretion or an impaired hormonal response to secreted insulin in the body [1–3]. Undiagnosed and poorly managed DM can progress to severe damage to organs in the body like the nerves, eyes, heart, and kidneys [4]. DM is divided into three forms based on clinical symptoms and aetiology [5]. These categorized forms of diabetes mellitus are Type I diabetes mellitus (TIDM), Type II diabetes mellitus (T2DM) and Gestational diabetes mellitus (GDM). Dietary habits and physical activity are major contributors to human health. Lifestyle and dietary changes over the past few decades have accelerated the incidence of diabetes in adults [6]. People adopt many modifications to their diets now a days due to food availability and cheaper prices. People tend to consume more high-fat, salt-loaded, and energy-dense foods [7]. Thus, by reducing obesity and high BMI through diet and lifestyle modifications, the burden of diabetes can be reduced [8]. According to the World Health Organization (WHO), 422 million people in the world suffer from diabetes and the mortality rate is 1.5. Prevalence mostly occurs in middle and low-income countries [9]. According to the National Diabetes Survey of Pakistan (NDSP 2016–17), 26.3% of the population has diabetes, which estimates 27.4 million people greater than 20 years of age have diabetes [10]. In Pakistan, 16.98% of the population had type 2 diabetes and 10.91% had pre-diabetes [11].

Probiotics usually termed as oral introduction of microorganisms in the body for certain beneficial effects. Most common probiotics used are Lactobacillus and Bifidobacterium [12–14]. For overall good health, gut microbiota should be in a balance state, any imbalance in gut micro biota causes many metabolic disorders such as insulin resistance due to production of metabolites and abnormality in utilization of nutrients [15, 16]. According to previous studies there is a significant difference noted in the intestinal microbiota of diabetics and controlled samples [17]. The differences in composition of the gut microbiota may affect immunity, transit time and permeability, inflammation and energy extracted from ingested food [18]. Magnesium is an important co-factor in many biochemical events, including about 300 enzymatic systems which regulate blood pressure, protein synthesis, muscle and neuron function and blood glucose regulation [19–21]. In order to treat diabetes and reduce the global burden of disease, dietary management can play a vital role [22]. A study conducted by Ling Jia et al., 2021 in School of Food Science and Technology, Jiangnan University, Wuxi, China researched the anti-diabetic properties of Clostridium Butyricum in mice [23]. They used 800 mg/kg/day sodium butyrate for 8 weeks and observed the lower Glycated haemoglobin (HbA1c) and fasting blood glucose when checked through oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) [23]. Another study of the human model was performed by Nadja Larsen et al. (2010) in the Department of Food Science, University of Copenhagen, and Frederiksberg, Denmark. They performed this study on 36 male adults divided into diabetic and non-diabetic male groups to identify changes in intestinal microorganisms via faces testing, investigated by tag-encoded Pyro sequencing and qPCR testing. They investigated that diabetes in adults had a strong association with compositional changes in intestinal microbiota and suggested that glucose tolerances should be considered important for diabetic treatment strategies through changes in microbiota [24]. In another study by Dae Jung Kim et al. (2010) in the Department of Nutrition, Gillings School of Global Public Health, and School of Medicine, the University of North Carolina conducted a 20-years follow-up study on 4,497 Americans (18–30 years of age) who had no diabetes at the start of the study. They studied the relationship of magnesium with diabetes, inflammatory markers and insulin resistance in subjects using the Coronary Artery Risk Development in Young Adults (CARDIA) study design. They found that magnesium intake had a significant inverse association with diabetes incidence in young adults. In order to determine the presence of diabetes in subjects, they use the hexokinase ultraviolet method, radioimmunoassay, Oral Glucose tolerance test, and Tosoh G-7 high performance liquid chromatography [25].

The numbers of patients with diabetes mellitus are increasing day by day and have caused several problems for human health and lowered the quality of life as compared to healthy and normal individuals. The current research work using magnesium supplementation and Clostridium butyricum as probiotics revealed a novel approach to curing type II diabetes mellitus because it enhanced the working efficiency of the gut microbiota and minimized the blood glucose level.

2.1 Induction of Diabetes

For induction of diabetes, 60 mg/kg body weight of Streptozotocin (STZ) is used in overnight fasting albino rats. After Streptozotocin induction, fasting blood glucose levels of > 200 mg/dl are considered diabetic in rats [26]. Rats were procured from the Animal House Institute of Molecular Biology and Biotechnology (IMBB) at the University of Lahore, Pakistan

2.2 Rat Bioassay

24 albino male rats were housed in the animal house of IMBB, The University of Lahore. Rats were kept on a standard basal diet for 1 week for acclimatisation purposes with controlled conditions like temperature (23 ± 2 ⁰C) and relative humidity (55%), along with a 12-hour light-dark period, and these conditions were followed throughout the trial.

A total of 24 rats were divided into four groups (n = 6). G₀ had 6 rats, which were diabetic control groups and fed a standard basal diet of metformin at 100 mg/kg/day [27, 28]. G₁ had six diabetic rats fed probiotic Clostridium Butyricum (2.5x10⁵ CFU/day) with a standard basal diet. G₂ had six diabetic rats fed magnesium (500 mg/kg) with a standard basal diet. G₃ had six diabetic rats given a combined probiotic (1.5x10⁵ CFU/day) and magnesium (300 mg/kg) with a standard basal diet [29, 30].

2.3 Glucose Assay

Random and fasting blood glucose levels were measured through the Glucometer device [11].

2.4 Magnesium Blood Test: Blood magnesium levels were measured by the Magnesium Assay Kit. A calmagite dye in the kit forms a colored complex specifically with magnesium. The intensity of the color, measured at 500 nm, is directly proportional to the magnesium concentration in the sample [20].

2.5 Microbial Assay: Fresh faces were collected from rat cages [31]. 30g of samples were diluted with 150 ml of sterile saline, homogenised in a standard blender, and filtered through gauze three times [32]. It was centrifuged at 6,000 rpm for 15 min, re-suspended in 150 ml of fresh sterile saline, and transferred directly to blood agar at a room temperature of 30°C in an anaerobic incubator [33]. After incubation at 37 ⁰C for 48 to 72 hours, colonies were identified on the basis of colony morphology [34].

2.6 Statistical Analysis

Data analysis was done using SPSS version 20. Quantitative variables like weight, blood glucose levels were represented as Mean ± SD. Differences in pre-treatment and post treatment variables like fasting and random blood glucose levels were analyzed using paired sample T-test.

Table 1, Pre and Post Treatment Mean ± SD values of Random Blood Glucose Levels of Albino Rats Induced Diabetic with Streptozotocin.

Week 1
Groups	Pre Treatment Mean ± SD	Post Treatment Mea n± SD	% Change	P value
G₀	206.6±67.7	246.1±17.1	19.1	0.203
G₁	206.6±67.7	189.3±32.6	8.4	0.431
G₂	231.7±101.3	255.2±34.3	10.1	0.534
G₃	126±7.07	213±7.07	69.0	0.429
Week 2
G₀	206.6±67.7	260.1±13.3	25.9	0.024
G₁	206.6±67.7	142.3±10.9	31.1	0.052
G₂	231.7±101.3	328.5±28.29	41.8	0.077
G₃	126±7.07	348.7±7.4	176.7	0.001
Week 3
G₀	206.6±67.7	370±22.3	79.1	0.005
G₁	206.6±67.7	85.1±3.8	58.8	0.006
G₂	231.7±101.3	354.2±17.6	52.9	0.061
G₃	126±7.07	288.5±7.07	129.0	0.008

Table 1 shows the random blood glucose levels of rats of all groups. Paired sample t-test was performed to observe the changes of treatment. Results are represented as mean value with standard deviation. In control group G₀, during week 1^stthere was no significant increase in random blood glucose level by 19.1%, during week 2^ndsignificant increase in random blood levels by 25.9% and at the end of week 3^rd significant elevated random blood glucose levels by 79.1%. In G₁, during week 1^st no significant reduction in random blood glucose levels were recorded and it was 8.4%, during week 2^ndsignificant reduction in blood glucose levels were observed as decrease in random blood glucose levels by 31.1%. At the end of week 3^rd there was significant decrease in random blood glucose by 58.8%. In G₂, during week 1^st there was no significant difference in random blood glucose levels (RBGL) by increase by 10.1%, while in week 2^nd and 3^rd there was no significant difference in blood glucose levels by increase in RBGL by 41.8% and 52.9% respectively. In G₃, there was no significant difference observed during week 1^st as increase in RBGL by 69%, while significant difference observed in week 2^nd by increase in RBGL by 176.7% and 3^rdby elevated RBGL by 129% respectively. The results showed that G₁ had significant reduction in RBGL by 8.4%, 31.1% and 58.8% respectively during the week wise treatment.

Table 2, Pre and Post Treatment Mean ± SD values of Fasting Blood Glucose Levels in Streptozotocin Induced Diabetic Rats

Group	Pre-Treatment Mean ± SD	Post-Treatment Mean ± SD	% Change	P value
G₀	142.6±12.2	261.6±17.9	83.5	0.000
G₁	101±2.68	72.33±16.8	28.4	0.004
G₂	115.0±2.3	232±124.7	101.7	0.152
G₃	140.5±6.36	239.5±7.7	70.5	0.006

Table 2 shows the mean values of fasting blood glucose levels for all groups’ pre and post treatment. Results are represented as mean values with standard deviation. In G₀ there was significant increase in fasting blood glucose levels (FBGL) by 83.5%, in G₁ there was signification reduction in fasting blood glucose levels (FBGL) by 28.4%, in G₂ there was increase in fasting blood glucose levels (FBGL) by 101.7%, while in G₃ there was increase in FBGL by 70.5%. These results showed that G₁ had significant reduction in fasting blood glucose levels (FBGL) by 28.4% at the end of treatment as shown in the Table 2

Table 3, Pre- and Post-Training Mean ± SD Values of Serum Magnesium in Streptozotocin-Induced Diabetic Rats.

Group	Pre Treatment Mean ± SD	Post Treatment Mean ± SD	% Change	P value
G₀	0.24±.00	0.27 ±.11	11.1	.256
G₁	0.28±.10	0.40 ±.15	30.0	.116
G₂	0.26±.12	0.38 ±.05	31.6	.049
G₃	0.27 ±.04	0.45 ±.15	40.0	.442

Table 3 shows the mean values of serum magnesium levels for all groups’ pre- and post-treatment. Results are represented as a mean value with a standard deviation. G₀showed no significant increase in serum magnesium levels by 11.1%, and G₁ expressed no significant increase in serum magnesium levels by 30.0%. G₂ showed a significant increase in serum magnesium levels by 31.1%, while in G₃, there was no significant increase in serum magnesium levels by 40.0%. These results showed that there was a significant increase in serum magnesium levels only in the magnesium-fed group (G₂), but all groups remained hypomagnesemia as mentioned in the Table 3.

Probiotics have a critical role in managing diabetes by affecting random blood glucose levels and improving gut function [35]. A study designed the placebo trial to observe the effect of magnesium on diabetes. 300 mg magnesium per day as magnesium sulphate for 3 months was given as compared to placebo, and they observed a positive effect of magnesium supplementation on random blood glucose levels from (239.1 ± 74.75 to 189.1 ± 60mg/dL versus 246.4 ± 97.37 to 247.8 ± 86.74 mg/dL, p < 0.01) [35], while contrary to our study, it showed no effect of magnesium supplementation on random blood glucose levels from 231.7 ± 101.3 to 354.2 ± 17.6 as a P value 0.06. The positive effect of magnesium on RBG levels could not be observed due to the shorter duration of treatment in this study or the use of magnesium oxide instead of magnesium sulphate [36]. Hani Al Salami et al., in 2008, experimented with this study to evaluate the effect of probiotics with gliclazide in diabetic rats, 40 rats were divided into 4 groups, and probiotic 75 mg/kg was given to rats. They observed random blood glucose levels decreasing by 2 folds (P < 0.01) in diabetic rats [37]. In this study, probiotic Clostridium butyricum also decreased RBG levels from 206.6 ± 67.7 to 85.1 ± 3.8 with a P value of 0.006. As Clostridium butyricum produced short chain fatty acids and butyrate producing bacteria which improves gut homeostasis and helps in controlling random blood glucose levels especially in diabetes [38]. Elevated fasting blood glucose level is characteristic sign of diabetes. Probiotic improves fasting blood glucose levels in diabetes [39]. A study was performed to access the hypoglycemic effect of Clostridium butyricum in rat trial where animals were divided into 2 groups and given Clostridium butyricum (2.5x10⁸) for 45 weeks, and observed significant results (P < 0.0172) in FBG levels. They also observed that Clostridium butyricum feeding at an early age delayed the onset of diabetes in rats [23]. The findings of this study also showed impressive effects in lowering FBG levels by the use of Clostridium butyricum. The results indicate that G₀ on a normal diet with metformin had no major impact on FBG levels from 142.6 ± 12.2 to 261.6 ± 17.9 (P value = 0.000), while G₁ with Clostridium butyricum treatment showed significant effects in lowering FBG levels from 101 ± 2.68 to 72.33 ± 16.8 (P value = 0.004). G₂ with magnesium treatment also had no such good impact on FBG levels, such as 115.0 ± 2.3 to 232 ± 124.7 values showing an increasing trend as P value = 0.152, which shows that magnesium had no effect in lowering blood glucose levels, instead of it magnesium in this study had no positive effects, magnesium solely caused bloating and constipation in rats. While G3, which was magnesium + probiotic, showed a significant increasing trend in FBG levels from 140.5 ± 6.36 to 239.5 ± 7.7 as P value = 0.006. In this study, it was hypothesized that solely magnesium supplementation through oral diet would reduce hyperglycemia, however the results showed that magnesium supplementation had no effect on lowering hyperglycemia, instead, it caused intestinal inflammation and paralytic ileus in rats, and 66.6% of rats died due to magnesium toxicity and hyperglycemia in this group [40]. G₀ showed a significant increase in serum magnesium levels from 0.24 ± .00 mg/dL to 0.27 mg/dL ± .11 observed as P = 0.256; G₁ expressed no significant increase in serum magnesium levels from 0.28 ± .10 mg/dL to 0.40 ± .15 mg/dL observed as P = 0.116. G₂ showed a significant increase in serum magnesium levels from 0.26 ± .12 mg/dL to 0.38 ± .05 mg/dL observed as P = 0.049, while in G₃ there was no significant increase in serum magnesium levels from 0.27 ± .04 mg/dL to 0.45 ± .15 mg/dL observed as P = 0.442. These results showed that there was a significant increase in serum magnesium levels only in the magnesium-fed group (G₂), but all groups remained hypomagnesemic. A study designed by Naila Masood et al. in 2009 observed the same effect that magnesium concentration had no significant difference in diabetes and the control group from 22.67 ± 24.5 mg/dL to 18.3 ± 3.4 mg/dL, P = 0.26 [41]. The positive effect of magnesium in controlling diabetes, which is described in various investigation, may not be observed in this study due to magnesium being given in the form of magnesium oxide, which caused constipation and bloating. As magnesium response is dose-dependent, the high dose of magnesium oxide in this study caused the problems stated above.

The use of Probiotic supplementation in diabetic control has many advantages. Probiotics improve gut flora, manage gut homeostasis, and reduce intestinal inflammation to improve insulin sensitivity [42]. Induction of Clostridium butyricum via oral administration in rats improved Clostridium butyricum load in gut microbiota composition. Ling Jia et al. in 2017 observed the same effect by conducting a study trail in which they divided rats into 2 groups and gave one group Clostridium butyricum for 5 weeks and the second group the same probiotic for 13 weeks, and they observed a significant change in the composition of Clostridium butyricum (P = 0.385.23) [23]. In this study, Clostridium butyricum changes were also observed via microbial analysis. Gut homeostasis plays a major role in managing metabolic diseases. The changes in the composition of Clostridium butyricum in each group indicate that G1 reported from Fig. 1 as of petri plate B, which had a greater number of Clostridium butyricum 10⁵ CFU/mL than other groups, had a significant impact on lowering random blood glucose levels (P value = 0.006), and an increase in weight was also observed in this group from 246.6 ± 8.6 to 258 ± 11.6 (P value = 0.001). The observations and results from G₀10³ CFU/mL, Fig. 1 from petri plate A indicates that a lower number of Clostridium butyricum was associated with a decrease in weight from 261.0 ± 8.08 to 197.7 ± 26.8 such as (P value = 0.036) and random blood glucose levels from 206.6 ± 67.7 to 370 ± 22.3 as the (P value = 0.005). In G2, from petri plate C, Fig. 1 the same trend of Clostridium butyricum was observed at 10³ CFU/ml, which was less than in G1, and results indicate that there was a significant decrease in weight from 251.3 ± 12.1 to 186.6 ± 13.8 as a (P value of 0.000), and the same trend of a lower load of Clostridium butyricum was observed in random blood glucose levels. There was increase in random blood glucose levels from 231.7 ± 101.3 to 328.5 ± 28.29 as (P value = 0.077), the same as the fasting blood glucose levels of G₂ from 115.0 ± 2.3 to 232 ± 124.7 as (P value = 0.152). G₃ petri plate D, Fig. 1, had a same relation of Clostridium butyricum load in fecal samples, observed as10⁴ CFU/ml, which was greater than G₂ and G₁ due to supplementation of Clostridium butyricum in combination with magnesium. Significant weight change occurred as 266 ± 7.07 to 181.2 ± 13.7 as (P value = 0.036)0, and random blood glucose levels as 126 ± 7.07 to 288.5 ± 7.07 as (P values = 0.008). Fasting blood glucose levels were observed at 140.5 ± 6.36 to 239.5 ± 7.7 with P value = 0.006. These results showed that Clostridium butyricum has major role in diabetes management as this probiotic produces short chain fatty acids which regulate intestinal homeostasis and help regulate blood glucose levels and increase growth performance [38].

The findings of this study reveal that, contrary to many other studies, magnesium didn’t show effective results in controlling blood glucose levels alone. However, Clostridium butyricum (2.5x10⁵ CFU)-fed rats showed great improvement in fasting and random blood glucose levels as compared to other groups, which were diabetic control and magnesium-fed groups. Clostridium butyricum improved gut health, boosted up the homeostasis and metabolism of the micro biota which was proved by microbial analysis of a Clostridium butyricum load of 105 CFU in stool samples of rats. Magnesium and Clostridium butyricum combination helped to overcome the type II diabetes as indicated by the results of our study. Further studies need to be carried out to evaluate the effect of magnesium oxide and the dose of magnesium in combination with Clostridium butyricum in diabetes for better and safer results.

Acknowledgments

We are highly grateful to the "Institute of Molecular Biology and Biotechnology", The University of Lahore, for providing their animal house. Xiao Zhang from the Clinical Research Centre of the Xuzhou Affiliated Hospital for providing assistance in carrying out research work.

Author Contributions Statement

UT, AA, and XZ, conceived the core ideas and designed the outlines of the manuscript. XZ contributed to the project administration and supervision. SJ, HR, and HK carried out the critical analysis of literature. MU and MB made significant contributions to write and revise the manuscript. XZ set-up the platforms, provided resources, and secured funding’s for the study. All authors read and approved the submitted version of the manuscript.

Funding:

The Unveiling & Leading Project of XZHMU under Grant No. JBGS202204. Xiao Zhang was supported by National Key Research and Development Program of China [No. 2020YFB1711500] and authors also express appreciation to the Deanship of Scientific Research at King Khalid University Saudi Arabia for funding through research groups program under grant number [R.G.P. 2/428/44].

Data Availability

Not applicable.

Ethics Approval and Consent to Participate

All methods were performed in accordance with the relevant guidelines and regulations. Written informed consent was obtained from all participants included in the study. This study was approved by Research Ethical Committee of University of Lahore: IRB-UOL-FAHS/822/2021.

Consent for publication

Not applicable.

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Publisher’s note

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.

N.G. Forouhi, N.J. Wareham, Epidemiology of diabetes, Medicine,38(11) (2010) 602-606.
M.F. Brereton, M. Iberl, K. Shimomura, Q. Zhang, A.E. Adriaenssens, P. Proks, I.I. Spiliotis, W. Dace, K.K. Mattis, R. Ramracheya, Reversible changes in pancreatic islet structure and function produced by elevated blood glucose, Nature communications,5(1) (2014) 4639.
A.D. Association, Diagnosis and classification of diabetes mellitus, Diabetes care,37(Supplement_1) (2014) S81-S90.
Z. Tao, A. Shi, J. Zhao, Epidemiological perspectives of diabetes, Cell biochemistry and biophysics,73 (2015) 181-185.
W. Kerner, J. Brückel, Definition, classification and diagnosis of diabetes mellitus, Experimental and clinical endocrinology & diabetes,122(07) (2014) 384-386.
A.R. Barnosky, K.K. Hoddy, T.G. Unterman, K.A. Varady, Intermittent fasting vs daily calorie restriction for type 2 diabetes prevention: a review of human findings, Translational Research,164(4) (2014) 302-311.
D.R. Jacobs Jr, Fast food and sedentary lifestyle: a combination that leads to obesity, Oxford University Press, 2006, pp. 189-190.
A.K. Jenum, I. Brekke, I. Mdala, M. Muilwijk, A. Ramachandran, M. Kjøllesdal, E. Andersen, K.R. Richardsen, A. Douglas, G. Cezard, Effects of dietary and physical activity interventions on the risk of type 2 diabetes in South Asians: meta-analysis of individual participant data from randomised controlled trials, Diabetologia,62 (2019) 1337-1348.
I.V. Kononenko, O.M. Smirnova, A.Y. Mayorov, M.V. Shestakova, Classification of diabetes. World Health Organization 2019. What’s new?, Diabetes mellitus,23(4) (2020) 329-339.
A. Basit, A. Fawwad, S.A. Siddiqui, K. Baqa, Current management strategies to target the increasing incidence of diabetes within Pakistan, Diabetes, metabolic syndrome and obesity: targets and therapy, (2018) 85-96.
A.H. Aamir, Z. Ul-Haq, S.A. Mahar, F.M. Qureshi, I. Ahmad, A. Jawa, A. Sheikh, A. Raza, S. Fazid, Z. Jadoon, Diabetes Prevalence Survey of Pakistan (DPS-PAK): prevalence of type 2 diabetes mellitus and prediabetes using HbA1c: a population-based survey from Pakistan, BMJ open,9(2) (2019) e025300.
R.J. Boyle, R.M. Robins-Browne, M.L. Tang, Probiotic use in clinical practice: what are the risks?, The American journal of clinical nutrition,83(6) (2006) 1256-1264.
H. Morita, H. Toh, K. Oshima, M. Murakami, T.D. Taylor, S. Igimi, M. Hattori, Complete genome sequence of the probiotic Lactobacillus rhamnosus ATCC 53103, Journal of bacteriology,191(24) (2009) 7630-7631.
B.T. Nguyen, E. Bujna, N. Fekete, A.T. Tran, J.M. Rezessy-Szabo, R. Prasad, Q.D. Nguyen, Probiotic beverage from pineapple juice fermented with Lactobacillus and Bifidobacterium strains, Frontiers in nutrition,6 (2019) 54.
F. Karlsson, V. Tremaroli, J. Nielsen, F. Bäckhed, Assessing the human gut microbiota in metabolic diseases, Diabetes,62(10) (2013) 3341-3349.
Y. Gálvez-Ontiveros, S. Páez, C. Monteagudo, A. Rivas, Endocrine disruptors in food: impact on gut microbiota and metabolic diseases, Nutrients,12(4) (2020) 1158.
A. Pascale, N. Marchesi, C. Marelli, A. Coppola, L. Luzi, S. Govoni, A. Giustina, C. Gazzaruso, Microbiota and metabolic diseases, Endocrine,61 (2018) 357-371.
A.W. Janssen, S. Kersten, The role of the gut microbiota in metabolic health, The FASEB Journal,29(8) (2015) 3111-3123.
K. Pasternak, J. Kocot, A. Horecka, Biochemistry of magnesium, Journal of Elementology,15(3) (2010) 601-616.
D.P. Chaudhary, R. Sharma, D.D. Bansal, Implications of magnesium deficiency in type 2 diabetes: a review, Biological trace element research,134 (2010) 119-129.
U. Gröber, J. Schmidt, K. Kisters, Magnesium in prevention and therapy, Nutrients,7(9) (2015) 8199-8226.
S.H. Ley, O. Hamdy, V. Mohan, F.B. Hu, Prevention and management of type 2 diabetes: dietary components and nutritional strategies, The Lancet,383(9933) (2014) 1999-2007.
L. Jia, D. Li, N. Feng, M. Shamoon, Z. Sun, L. Ding, H. Zhang, W. Chen, J. Sun, Y.Q. Chen, Anti-diabetic effects of Clostridium butyricum CGMCC0313. 1 through promoting the growth of gut butyrate-producing bacteria in type 2 diabetic mice, Scientific reports,7(1) (2017) 7046.
N. Larsen, F.K. Vogensen, F.W. Van Den Berg, D.S. Nielsen, A.S. Andreasen, B.K. Pedersen, W.A. Al-Soud, S.J. Sørensen, L.H. Hansen, M. Jakobsen, Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults, PloS one,5(2) (2010) e9085.
D.J. Kim, P. Xun, K. Liu, C. Loria, K. Yokota, D.R. Jacobs Jr, K. He, Magnesium intake in relation to systemic inflammation, insulin resistance, and the incidence of diabetes, Diabetes care,33(12) (2010) 2604-2610.
J.N. Asiwe, K.K. Anachuna, E.G. Moke, K.O. Sanusi, D.E. Okonofua, O. Omeru, A.A. Fasanmade, High dietary salt intake alleviates fasting blood glucose in streptozotocin-induced diabetic male Wistar rats, The Thai Journal of Pharmaceutical Sciences,45(3) (2021) 172-177.
M. Pellizzon, Choice of laboratory animal diet influences intestinal health, Lab animal,45(6) (2016) 238.
O. Fapohunda, O. Balogun, Oral magnesium supplementation modulates hepatic and intestinal expression of some carbohydrate metabolizing genes in type 2 diabetic rats, Int J Mol Biol Open Access,4(6) (2019) 189-194.
C.-C. Lin, G.-J. Tsweng, C.-F. Lee, B.-H. Chen, Y.-L. Huang, Magnesium, zinc, and chromium levels in children, adolescents, and young adults with type 1 diabetes, Clinical Nutrition,35(4) (2016) 880-884.
C. Chu, W. Zhao, Y. Zhang, L. Li, J. Lu, L. Jiang, C. Wang, W. Jia, Low serum magnesium levels are associated with impaired peripheral nerve function in type 2 diabetic patients, Scientific reports,6(1) (2016) 32623.
C.H. Lee, T. Steiner, E.O. Petrof, M. Smieja, D. Roscoe, A. Nematallah, J.S. Weese, S. Collins, P. Moayyedi, M. Crowther, Frozen vs fresh fecal microbiota transplantation and clinical resolution of diarrhea in patients with recurrent Clostridium difficile infection: a randomized clinical trial, Jama,315(2) (2016) 142-149.
E. Mattila, R. Uusitalo–Seppälä, M. Wuorela, L. Lehtola, H. Nurmi, M. Ristikankare, V. Moilanen, K. Salminen, M. Seppälä, P.S. Mattila, Fecal transplantation, through colonoscopy, is effective therapy for recurrent Clostridium difficile infection, Gastroenterology,142(3) (2012) 490-496.
M.J. Hamilton, A.R. Weingarden, M.J. Sadowsky, A. Khoruts, Standardized frozen preparation for transplantation of fecal microbiota for Recurrentclostridium difficileinfection, Official journal of the American College of Gastroenterology| ACG,107(5) (2012) 761-767.
N.G. Rossen, S. Fuentes, M.J. van der Spek, J.G. Tijssen, J.H. Hartman, A. Duflou, M. Löwenberg, G.R. van den Brink, E.M. Mathus-Vliegen, W.M. de Vos, Findings from a randomized controlled trial of fecal transplantation for patients with ulcerative colitis, Gastroenterology,149(1) (2015) 110-118. e4.
S. Singh, R. Sharma, S. Malhotra, R. Pothuraju, U. Shandilya, Lactobacillus rhamnosus NCDC17 ameliorates type-2 diabetes by improving gut function, oxidative stress and inflammation in high-fat-diet fed and streptozotocintreated rats, Beneficial microbes,8(2) (2017) 243-255.
M. Solati, E. Ouspid, S. Hosseini, N. Soltani, M. Keshavarz, M. Dehghani, Oral magnesium supplementation in type II diabetic patients, Medical journal of the Islamic Republic of Iran,28 (2014) 67.
H. Al-Salami, G. Butt, J.P. Fawcett, I.G. Tucker, S. Golocorbin-Kon, M. Mikov, Probiotic treatment reduces blood glucose levels and increases systemic absorption of gliclazide in diabetic rats, European journal of drug metabolism and pharmacokinetics,33 (2008) 101-106.
Q. Zhang, X. Xiao, M. Li, M. Yu, F. Ping, J. Zheng, T. Wang, X. Wang, Vildagliptin increases butyrate-producing bacteria in the gut of diabetic rats, PloS one,12(10) (2017) e0184735.
M. Babashahi, M. Mirlohi, R. Ghiasvand, L. Azadbakht, L. Mosharaf, S. Torki-Baghbadorani, Effects of probiotic soy milk fermented by lactobacillus plantarum A7 (KC 355240) added with Cuminum Cyminum essential oil on fasting blood glucose levels, serum lipid profile and body weight in diabetic Wistar rats, International Journal of Preventive Medicine,11 (2020).
J. Golzarian, H. William Scott, W.O. Richards, Hypermagnesemia-induced paralytic ileus, Digestive diseases and sciences,39 (1994) 1138-1142.
N. Masood, G.H. Baloch, R.A. Ghori, I.A. Memon, M.A. Memon, M.S. Memon, Serum zinc and magnesium in type-2 diabetic patients, J Coll Physicians Surg Pak,19(8) (2009) 483-486.
A. H Rad, S. Abbasalizadeh, S. Vazifekhah, F. Abbasalizadeh, T. Hassanalilou, P. Bastani, H.-S. Ejtahed, A.-R. Soroush, M. Javadi, A. M Mortazavian, The future of diabetes management by healthy probiotic microorganisms, Current diabetes reviews,13(6) (2017) 582-589.

No competing interests reported.

Download PDF

Journal Publication

published 01 Apr, 2024

Read the published version in BMC Microbiology →

Editorial decision: Major revision
14 Sep, 2023
Editor assigned by journal
14 Sep, 2023
Submission checks completed at journal
14 Sep, 2023
First submitted to journal
09 Sep, 2023

You are reading this latest preprint version

Restoration of gut dysbiosis through Clostridium butyricum and magnesium possibly balance blood glucose levels: an experimental study

Status:

Journal Publication

Version 1

Abstract

Figures

1. Introduction

2. Materials & Methods

2.1 Induction of Diabetes

2.2 Rat Bioassay

2.3 Glucose Assay

3. Results

4. Discussion

5. Conclusion

Declarations

References

Additional Declarations

Status:

Journal Publication

Version 1