Aim Butyrate is a major gut microbiota‐derived metabolite. Reduced butyrate‐producing bacteria has been reported in the spontaneously hypertensive rat (SHR), a model of hypertension characterized by dysfunctional autonomic nervous system and gut dysbiosis. Here, we demonstrate a potential mechanism for butyrate in blood pressure regulation. Methods High‐performance liquid chromatography and liquid chromatography‐mass spectrometry were performed to measure butyrate levels in feces and serum. Ussing chamber determined butyrate transport in colon ex vivo. Real‐time PCR and immunohistochemistry evaluated expression of butyrate transporter, Slc5a8, in the colon. Mean arterial blood pressure was measured in catheterized anesthetized rats before and after a single butyrate intracerebroventricular injection. Activity of cardioregulatory brain regions was determined by functional magnetic resonance imaging to derive neural effects of butyrate. Results In the SHR, we demonstrated elevated butyrate levels in cecal content, but diminished butyrate levels in circulation, possibly due to reduced expression of Slc5a8 transporter in the colon. In addition, we observed lower expression levels of butyrate‐sensing receptors in the hypothalamus of SHR, likely leading to the reduced effects of centrally administered butyrate on blood pressure in the SHR. Functional magnetic resonance imaging revealed reduced activation of cardioregulatory brain regions following central administration of butyrate in the SHR compared to control. Conclusion We demonstrated a reduced availability of serum butyrate in the SHR, possibly due to diminished colonic absorption. Reduced expression of butyrate‐sensing receptors in the SHR hypothalamus may explain the reduced central responsiveness to butyrate, indicating microbial butyrate may play a role in blood pressure regulation.

The brain-gut axis plays a critical role in the regulation of different diseases, many of which are characterized by sympathetic dysregulation. However, a direct link between sympathetic dysregulation and gut dysbiosis remains to be illustrated. Bone marrow (BM)-derived immune cells continuously interact with the gut microbiota to maintain homeostasis in the host. Their function is largely dependent upon the sympathetic nervous system acting via adrenergic receptors present on the BM immune cells. In this study, we utilized a novel chimera mouse that lacks the expression of BM beta1/2 adrenergic receptors (b1/2-ARs) to investigate the role of the sympathetic drive to the BM in gut and microbiota homeostasis. Fecal analyses demonstrated a shift from a dominance of Firmicutes to Bacteroidetes phylum in the b1/2-ARs KO chimera, resulting in a reduction in Firmicutes/Bacteroidetes ratio. Meanwhile, a significant reduction in Proteobacteria phylum was determined. No changes in the abundance of acetate-, butyrate-, and lactate-producing bacteria, and colon pathology were observed in the b1/2-ARs KO chimera. Transcriptomic profiling in colon identified Killer Cell Lectin-Like Receptor Subfamily D, Member 1 (Klrd1), Membrane-Spanning 4-Domains Subfamily A Member 4A (Ms4a4b), and Casein Kinase 2 Alpha Prime Polypeptide (Csnk2a2) as main transcripts associated with the microbiota shifts in the b1/2-ARs KO chimera. Suppression of leukocyte-related transcriptome networks (i.e., function, differentiation, migration), classical compliment pathway, and networks associated with intestinal function, barrier integrity, and excretion was also observed in the colon of the KO chimera. Moreover, reduced expression of transcriptional networks related to intestinal diseases (i.e., ileitis, enteritis, inflammatory lesions, and stress) was noted. The observed suppressed transcriptome networks were associated with a reduction in NK cells, macrophages, and CD4+ T cells in the b1/2-ARs KO chimera colon. Thus, sympathetic regulation of BM-derived immune cells plays a significant role in modifying inflammatory networks in the colon and the gut microbiota composition. To our knowledge, this study is the first to suggest a key role of BM b1/2-ARs signaling in host-microbiota interactions, and reveals specific molecular mechanisms that may lead to generation of novel anti-inflammatory treatments for many immune and autonomic diseases as well as gut dysbiosis across the board.

Emerging evidence suggests an associative link between gut dysbiosis, the autonomic nervous system (ANS) and the immune system in pathophysiology of neurogenic hypertension (HTN). However, the close interplay between these three systems presents us with difficulties in deciphering the cause-effect relationship in disease. The present study utilized beta 1 and 2 adrenergic receptor knock out (AdrB1tm1BkkAdrB2tm1Bkk/J KO) mice to isolate the effects of reduced overall sympathetic drive on gut microbiota and systemic immune system. We observed the following: (i) Diminished beta adrenergic signaling mainly reflects in shifts in the Firmicutes phyla, with a significant increase in abundance of largely beneficial Bacilli Lactobacillales in the KO mice; (ii) This was associated with increased colonic production of beneficial short chain fatty acids (SCFAs) butyrate, acetate and propionate, confirming functional microbiota shifts in the KO mice; (iii) Dampened systemic immune responses in the KO mice reflected in reduction on circulating CD4+.IL17+ T cells and increase in young neutrophils, both previously associated with shifts in the gut microbiota. Taken together, these observations demonstrate that reduced expression of beta adrenergic receptors may lead to beneficial shifts in the gut microbiota and dampened systemic immune responses. Considering the role of both in hypertension, this suggests that dietary intervention may be a viable option for manipulation of blood pressure via correcting gut dysbiosis.

Gut microbiota is crucial for function of the gastrointestinal (GI) tract, and modulates the communication between the GI tract and the central nervous system. Microbial metabolites such as propionate have been shown to regulate blood pressure (BP), while butyrate, one of the major bacterial fermented byproducts, reportedly produces beneficial effects in multiple dysbiosis-related diseases. We previously showed that chronic supplementation with soluble fiber-rich, butyrolytic diet modified microbiota and increased BP in the spontaneously hypertensive rat (SHR). Here, we tested the impact of same diet on BP regulation in the Wistar Kyoto rats (WKY). Methods: Male 4 weeks old WKY were placed on either the fructooligosaccharides/inulin-rich diet (fiber, N=6), or its calorie-matched control diet (control, N=6) (Research Diets, Inc.) for 14 weeks. Baseline BP was measured by tail cuff every week for the duration of the study, and by telemetry at the end of the study and during 20 minutes of restraint stress. Spectral analysis of BP waveform was performed during restraint stress to measure autonomic variables. Results: We observed no significant difference in mean BP measured by tailcuff or by telemetry between the two groups. However, there was an increase in mean BP after 2 minutes of restraint (104.5 ± 7.45 mmHg vs. 116.3 ± 3.19 mmHg; p=0.0411, n=6), which was associated with a trend in increase in LF/HF variable linked with vasovagal balance (1.99 ± 1.43 vs. 3.04 ± 1.46, p=0.17) in the fiber group when submitted to restraint stress. Conclusion: Unlike in the SHR, fiber-rich diet did not alter baseline BP in the normotensive WKY. However, chronic fiber diet produced autonomic imbalance and increased BP in response to restraint stress in the WKY.

Introduction: The link between gut dysbiosis and hypertension is characterized by a significant reduction in butyrate-producing bacteria in the spontaneously hypertensive rat (SHR) compared with its Wistar Kyoto (WKY) control. Butyrate, a major end-product of gut microbiota fermentation, has beneficial effects in multiple dysbiosis-related diseases. Here, we investigated the link between serum butyrate levels and central effects of butyrate in regulation of blood pressure (BP) in WKY and SHR. Methods: Serum samples were collected from 12 weeks old male WKY and SHR, and butyrate quantification was performed using LC-MS. Quantitative real time PCR was employed to measure relative expression levels of butyrate sensing receptors (Gpr41, Gpr43) in the hypothalamus of SHR and WKY. Central effects of butyrate on BP were evaluated by acute ICV injection in anaesthetized WKY and SHR. Results: Significantly lower levels of serum butyrate were observed in the SHR (WKY 1.23±0.14μmol/L vs SHR 0.23±0.033μmol/L, N=4, P=0.0005). Moreover, lower expression of Gpr43 were observed in the hypothalamus of SHR (WKY 0.0002 ± 2.7e-005 vs SHR 6.9e-005 ± 1.9e-005, N=4/3, P=0.0152). ICV administration of butyrate reduced BP in both strains; however, the reduction in arterial BP was greater in the WKY at two time points (400s post-injection: WKY -5.83±0.49mmHg vs SHR -2.62±0.63mmHg, N=4, P=0.0235; 1000s post-injection: WKY -19.7±1.85mmHg vs SHR -13.26±1.1mmHg, N=3, P=0.05). Conclusion: Reduced serum butyrate is associated with reduced sensitivity to central butyrate. This may contribute to elevation in BP in the SHR.