Korean J Physiol Pharmacol.  2017 Jan;21(1):1-9. 10.4196/kjpp.2017.21.1.1.

Ameliorative effects of atractylodin on intestinal inflammation and co-occurring dysmotility in both constipation and diarrhea prominent rats

• Yu C
• Xiong Y
• Chen D
• Li Y
• Xu B
• Lin Y
• Tang Z
• Jiang C
• Wang L

• KMID: 2371062
• DOI: http://doi.org/10.4196/kjpp.2017.21.1.1

Abstract

Intestinal disorders often co-occur with inflammation and dysmotility. However, drugs which simultaneously improve intestinal inflammation and co-occurring dysmotility are rarely reported. Atractylodin, a widely used herbal medicine, is used to treat digestive disorders. The present study was designed to characterize the effects of atractylodin on amelioration of both jejunal inflammation and the co-occurring dysmotility in both constipation-prominent (CP) and diarrhea-prominent (DP) rats. The results indicated that atractylodin reduced proinflammatory cytokines TNF-α, IL-1β, and IL-6 in the plasma and inhibited the expression of inflammatory mediators iNOS and NF-kappa B in jejunal segments in both CP and DP rats. The results indicated that atractylodin exerted stimulatory effects and inhibitory effects on the contractility of jejunal segments isolated from CP and DP rats respectively, showing a contractile-state-dependent regulation. Atractylodin-induced contractile-state-dependent regulation was also observed by using rat jejunal segments in low and high contractile states respectively (5 pairs of low/high contractile states). Atractylodin up-regulated the decreased phosphorylation of 20 kDa myosin light chain, protein contents of myosin light chain kinase (MLCK), and MLCK mRNA expression in jejunal segments of CP rats and down-regulated those increased parameters in DP rats. Taken together, atractylodin alleviated rat jejunal inflammation and exerted contractile-state-dependent regulation on the contractility of jejunal segments isolated from CP and DP rats respectively, suggesting the potential clinical implication for ameliorating intestinal inflammation and co-occurring dysmotility.

Keyword

Atractylodin; Constipation; Contractile-state-dependent regulation; Diarrhea; Inflammation; Jejunal contractility

MeSH Terms

Animals
Constipation*
Cytokines
Diarrhea*
Herbal Medicine
Inflammation*
Interleukin-6
Myosin Light Chains
Myosin-Light-Chain Kinase
NF-kappa B
Phosphorylation
Plasma
Rats*
RNA, Messenger
Cytokines
Interleukin-6
Myosin Light Chains
Myosin-Light-Chain Kinase
NF-kappa B
RNA, Messenger

Figure

• 

  Fig. 1 Atractylodin-exerted contractile-state-dependent regulation on rat jejunal contractility.Representative traces of atractylodin-exerted stimulatory and inhibitory effects on rat jejunal contractility in CP and DP rats, respectively. Contractile amplitude of jejunal segments in normal contractile state is set to a relative value of 100% (control). Other data are the relative values compared with control. **p<0.01 compared with the NC; ##p<0.01 compared with contractile amplitude of jejunal segments in DP group or CP group before being given atractylodin.

• 

  Fig. 2 Histomorphological characteristics of atractylodin-induced anti-inflammatory effects on DP and CP rats.Panel A: the histology images including normal jejunal tissue, CP jejunal tissue, DP jejunal tissue, CP+Atrac jejunal tissue, and DP+Atrac jejunal tissue. Panel B: statistical analysis of the effects of atractylodin on the serum contents of TNF-α, IL-1β and IL-6 in both DP and CP rats. Data are expressed as mean±SD (% NC, n=6); **p<0.01 compared with the normal controls; ##p<0.01 compared with CP group or DP group before being given Atractylodin, respectively. Atrac, atractylodin; NC, normal control rats; CP, constipation prominent rats; DP, diarrhea prominent rats.

• 

  Fig. 3 Effect of atractylodin on iNOS and NF-κB in CP and DP rats.Panel A: histology images of iNOS expression (red fluorescence), DAPI nuclear staining (blue fluorescence), and merged images of the immunofluorescence of iNOS and DAPI. Panel B: histology images of NF-κB expression, DAPI nuclear staining, and merged images of the immunofluorescence of NF-κB and DAPI. DAPI, 4′,6-diamidino-2-phenylindole. Cells viewed at ×200 magnification.

• 

  Fig. 4 The characteristic of atractylodin-exerted contractile state dependent contractile-state-dependent regulation.Panel A: Representative traces (A1) and statistical analysis (A2) of atractylodin-induced stimulatory effects on the contractility of jejunal segment in 5 low contractile states. Panel B: Representative traces (B1) and statistical analysis (B2) of atractylodin-induced inhibitory effects on the contractility of jejunal segment in 5 high contractile states. Contractile amplitude of jejunal segments in normal contractile state is set to a relative value of 100% ( normal control, NC). Other data are the relative values compared with NC. Data are expressed as mean±SD (% NC, n=6); **p<0.01 compared with the NC; ##p<0.01 compared with contractile amplitude of jejunal segments in LCS (low contractile state) or HCS (high contractile state) before being given atractylodin, respectively. NC, normal control; SNP, sodium nitroprusside; AD, adrenaline; ACh, acetylcholine; ERY, erythromycin.

• 

  Fig. 5 Effects of atractylodin on myosin phosphorylation-related mechanisms.Panel A: phosphorylation extent (a1) and statistical analysis (a2) of 20-kDa myosin light chain (p-MLC20), Panel B: protein contents (b1) and statistical analysis (b2) of myosin light chain kinase (MLCK), Panel C: MLCK mRNA expression in jejunal segment in normal control (NC) rats, atractylodin-treated CP rats (CP+Atrac), and atractylodin-treated DP rats (DP+Atrac). Data were expressed as the mean±SD from four independent experiments; **p<0.01 compared with the normal controls; ##p<0.01 compared with CP group or DP group before being given atractylodin, respectively. p-MLC20, phosphorylated 20-kDa myosin light chain; MLC20, 20-kDa myosin light chain; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

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